Recombinant host cells comprising phosphoketolases
09550999 ยท 2017-01-24
Assignee
Inventors
- Michael Dauner (Claymont, DE)
- Lori Ann Maggio-Hall (Wilmington, DE)
- Jean-Francois Tomb (Wilmington, DE)
Cpc classification
Y02E50/10
GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
C12N15/74
CHEMISTRY; METALLURGY
C12P7/46
CHEMISTRY; METALLURGY
C12P13/08
CHEMISTRY; METALLURGY
Y02P20/52
GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
C12N9/0008
CHEMISTRY; METALLURGY
C12P13/06
CHEMISTRY; METALLURGY
C12Y102/07001
CHEMISTRY; METALLURGY
C12N9/1029
CHEMISTRY; METALLURGY
C12Y203/01054
CHEMISTRY; METALLURGY
International classification
C12P7/46
CHEMISTRY; METALLURGY
C12P13/06
CHEMISTRY; METALLURGY
C12P13/08
CHEMISTRY; METALLURGY
C12N1/00
CHEMISTRY; METALLURGY
C12N9/00
CHEMISTRY; METALLURGY
C12N15/00
CHEMISTRY; METALLURGY
C12N15/74
CHEMISTRY; METALLURGY
Abstract
The present invention is related to recombinant host cells comprising: (i) at least one deletion, mutation, and/or substitution in an endogenous gene encoding a polypeptide that converts pyruvate to acetaldehyde, acetyl-phosphate, or acetyl-CoA; and (ii) a heterologous polynucleotide encoding a polypeptide having phosphoketolase activity. The present invention is also related to recombinant host cells further comprising (iii) a heterologous polynucleotide encoding a polypeptide having phosphotransacetylase activity.
Claims
1. A recombinant host cell comprising: (i) at least one genetic modification in an endogenous gene encoding a polypeptide having pyruvate decarboxylase activity, wherein the at least one genetic modification reduces or eliminates pyruvate decarboxylase activity; and (ii) a heterologous polynucleotide encoding a polypeptide having phosphoketolase activity.
2. The recombinant host cell of claim 1, wherein said host cell further comprises: (iii) a heterologous polynucleotide encoding a polypeptide having phosphotransacetylase activity.
3. The recombinant host cell of claim 1, wherein said host cell has a reduced requirement for an exogenous two-carbon substrate for its growth in culture compared to a recombinant eukaryotic host cell comprising (i) and not (ii).
4. The recombinant host cell of claim 1, wherein said host cell has improved growth as compared to a host cell comprising (i) and not (ii) in culture media that is not supplemented with an exogenous two-carbon carbon substrate.
5. The recombinant host cell of claim 4, wherein said exogenous to-carbon substrate is ethanol or acetate.
6. The recombinant host cell of claim 1, wherein said endogenous gene encoding a polypeptide having pyruvate decarboxylase activity is PDC1, PDC5, PDC6, or combinations thereof and said host cell is S. cerevisiae.
7. The recombinant host cell of claim 1, wherein said heterologous polynucleotide encoding a polypeptide having phosphoketolase activity is xpk1 from Lactobacillus plantarum, xpkA from Lactobacillus pentosus MD363 or 6-phosphate phosphoketolase from B. lactis.
8. The recombinant host cell of claim 1 wherein said polypeptide having phosphoketolase activity comprises at least 85% identity to SEQ ID NO 481 or an active fragment thereof.
9. The recombinant host cell of claim 2, wherein said heterologous polynucleotide encoding a polypeptide having phosphotransacetylase activity is EutD from Lactobacillus plantarum or phosphotransacetylase from Bacillus subtilis.
10. The recombinant host cell of claim 2, wherein said, polypeptide having phosphotransacetylase activity comprises at least 85% identity to SEQ ID NO: 1472 or an active fragment thereof.
11. The recombinant host cell of claim 1, wherein said host cell is a member of the genera Clostridium, Zymomonas, Escherichia, Salmonella, Serratia, Erwinia, Klebsiella, Shigella, Rhodococcus, Pseudomonas, Bacillus, Lactobacillus, Enterococcus, Alcaligenes, Paenibacillus, Arthrobacter, Corynebacterium, Brevibacterium, Schizosaccharomyces, Kluyveromyces, Yarrowia, Pichia, Candida, Hansenula, or Saccharomyces.
12. The recombinant host cell of claim 1 wherein said host, cell is Saccharomyces cerevisiae.
13. The recombinant host cell of claim 1 further comprising a pyruvate-utilizing biosynthetic pathway, wherein said pyruvate-utilizing biosynthetic pathway forms a product selected from the group consisting of: 2,3-butanediol, 2-butanol, 2-butanone, valine, leucine, lactic acid, malate, and isoamyl alcohol.
14. The recombinant host cell of claim 1, further comprising a pyruvate-utilizing biosynthetic pathway herein said pyruvate-utilizing biosynthetic pathway is an isobutanol biosynthetic pathway comprising the substrate to product conversions: (i) pyruvate to acetolactate; (ii) acetolactate to 2,3-dihydroxyisovalerate; (iii) 2,3-dihydroxyisovalerate to 2-ketoisovalerate; (iv) 2-ketoisovalerate to isobutyraldehyde; (v) isobutyraldehyde to isobutanol; and wherein said recombinant host cell produces isobutanol.
15. The recombinant host cell of claim 13, wherein said pyruvate-utilizing biosynthetic pathway is a 2-butanone biosynthetic pathway comprising the substrate to product conversions: (i) pyruvate to acetolactate; (ii) acetolactate to acetoin; (iii) acetoin to 2,3-butanediol; (iv) 2,3-butanediol to 2-butanone; wherein said recombinant host cell produces 2-butanone.
16. The recombinant host cell of claim 13, wherein said pyruvate-utilizing biosynthetic pathway is a 2-butanol biosynthetic pathway comprising the substrate to product conversions: (i) pyruvate to acetolactate; (ii) acetolactate to acetoin; (iii) acetoin to 2,3-butanediol; (iv) 2,3-butanediol to 2-butanone; (v) 2-butanone to 2-butanol; wherein said recombinant host cell produces 2-butanol.
17. The recombinant host cell of claim 1 wherein the phosphoketolase matches the Profile HMM given in Table 6 with an E value of less than 7.5E-242.
18. The recombinant host cell of claim 1 wherein the phosphoketolase matches the Profile HMMs given in Tables 6, 7, 8, and 9 with E values of less than 7.5E-242, 1.1E-124, 2.1E-49, 7.8E-37, respectively.
19. The recombinant host cell of claim 1 further comprising a phosphotransacetylase which matches the Profile HMM given in Table 14 with an E value of less than 5E-34.
20. The recombinant host cell of claim 13, wherein said pyruvate-utilizing biosynthetic pathway is a 2,3-butanediol biosynthetic pathway comprising the substrate to product conversations: (i) pyruvate to acetolactate; (ii) acetolactate to acetoin; (iii) acetoin 2,3-butanediol; and wherein said recombinant host cell produces 2,3 butanediol.
21. A method for the production of 2,3-butanediol, 2-butanol, 2-butanone, valine, leucine, lactic acid, malic acid, alanine, fumaric acid, succinic acid or isoamyl alcohol comprising growing the recombinant host cell of claim 13 under conditions wherein the product is produced and optionally recovering the product.
22. A method for the production of isobutanol comprising growing the recombinant host cell of claim 14 under conditions wherein the product is produced and optionally recovering the product.
Description
BRIEF DESCRIPTION OF THE DRAWINGS/FIGURES AND INCORPORATION OF SEQUENCE LISTING AND TABLES
(1) The various embodiments of the invention can be more fully understood from the detailed description, the figures, and the accompanying sequence descriptions, which form a part of this application.
(2)
(3)
(4)
(5)
(6)
(7)
(8)
(9)
(10) Tables 6, 7, 8, 9, and 14 are tables of the Profile HMMs described herein. Table 6, 7, 8, and 14 are submitted herewith electronically and are incorporated herein by reference.
(11) The invention can be more fully understood from the following detailed description and the accompanying sequence descriptions which form a part of this application.
(12) The sequence listing provided herewith is herein incorporated by reference and conforms with 37 C.F.R. 1.821-1.825 (Requirements for Patent Applications Containing Nucleotide Sequences and/or Amino Acid Sequence Disclosuresthe Sequence Rules) and is consistent with World Intellectual Property Organization (WIPO) Standard ST.25 (2009) and the sequence listing requirements of the EPO and PCT (Rules 5.2 and 49.5 (a-bis), and Section 208 and Annex C of the Administrative Instructions). The symbols and format used for nucleotide and amino acid sequence data comply with the rules set forth in 37 C.F.R. 1.822. The content of the electronically submitted sequence listing Name: 20110615_CL4871USNA_SeqList.txt; Size: 6.67 MB; and Date of Creation/Modification: Jun. 9, 2011/Jun. 15, 2011 is incorporated herein by reference in its entirety.
(13) SEQ ID NOs: 1-20 are sequences of PDC target gene coding regions and proteins.
(14) SEQ ID NOs: 21-638 are phosphoketolase target gene coding regions and proteins.
(15) SEQ ID NOs: 762-1885 are phosphotransacetylase target gene coding regions and proteins.
(16) SEQ ID NOs: 1893-1897 are hybrid promoter sequences.
(17) SEQ ID NOs: 639-642, 644-654, 656-660, 662-701-714, 725-726, 729-740, 742-748, and 750-761 are primers.
(18) SEQ ID NO: 643 is the vector pRS426::GPD-xpk1+ADH1-eutD.
(19) SEQ ID NO: 655 is the TEF1p-kan-TEF1t gene.
(20) SEQ ID NO: 661 is vector pLA54.
(21) SEQ ID NO: 715 is vector pRS423::pGAL1-cre.
(22) SEQ ID NO: 716 is the vector pLH468-sadB.
(23) SEQ ID NOs: 717 and 718 are the amino acid and nucleic acid sequences for sadB from Achromobacter xylosoxidans.
(24) SEQ ID NO: 719 is the kivD coding region from L. lactis.
(25) SEQ ID NO: 720 is the plasmid pRS425::GPM-sadB.
(26) SEQ ID NO: 721 is the GPM promoter.
(27) SEQ ID NO: 722 is the ADH1 terminator.
(28) SEQ ID NO: 723 is the GPM-sadB-ADHt segment.
(29) SEQ ID NO: 724 is the pUC19-URA3 plasmid.
(30) SEQ ID NO: 741 is the ilvD-FBA1t segment.
(31) SEQ ID NO 749 is URA3r2 template DNA.
(32) SEQ ID NO: 1886 is the ilvD coding region from S. mutans.
(33) SEQ ID NO: 1888 is vector 011468.
(34) SEQ ID NO: 1898 is pUC19-URA3::pdc1::GPD-xpk1+ADH1-eutD.
(35) SEQ ID NOs: 1899-1906 are the sequences of modified S. cerevisiae loci.
(36) SEQ ID NO: 1907 is the sequence of pLH702.
(37) SEQ ID NO: 1908 is the sequence of pYZ067DkivDDhADH
(38) SEQ ID NO: 1909 is the amino acid sequence of ALD6.
(39) SEQ ID NO: 1910 is the amino acid sequence of K9D3.
(40) SEQ ID NO: 1911 is the amino acid sequence of K9G9.
(41) SEQ ID NO: 1912 is the amino acid sequence of YMR226c.
(42) SEQ ID NOs: 1913 and 1914 are the nucleic acid and amino acid sequences of AFT1.
(43) SEQ ID NOs: 1915 and 1916 are the nucleic acid and amino acid sequences of AFT2.
(44) SEQ ID NOs: 1917 and 1918 are the nucleic acid and amino acid sequences of FRA2.
(45) SEQ ID NOs. 1919 and 1920 are the nucleic acid and amino acid sequences of GRx3.
(46) SEQ ID NOs: 1921 and 1922 are the nucleic acid and amino acid sequences of CCC1.
(47) SEQ ID NO: 1923 is the amino acid sequence of an alcohol dehydrogenase from Beijerinkia indica.
DETAILED DESCRIPTION OF THE INVENTION
(48) Applicants have solved the stated problem by reducing or eliminating the need for providing two substrates, one of which is converted to a desired product, the other fully or partly into acetyl-CoA by recombinant host cells requiring such supplementation for growth comprising the expression of enzymes of the phosphoketolase pathway in such cells. One such enzyme, phosphoketolase (Enzyme Commission Number EC 4.1.2.9), catalyzes the conversion of xylulose 5-phosphate into glyceraldehyde 3-phosphate and acetyl-phosphate (Heath et al., J. Biol. Chem. 231: 1009-29; 1958). Another such enzyme is phosphotransacetylase (Enzyme Commission Number EC 2.3.1.8) which converts acetyl-phosphate into acetyl-CoA.
(49) Applicants have provided PDC-KO recombinant host cells comprising a heterologous polynucleotide encoding a polypeptide having phosphoketolase activity, and optionally a heterologous polynucleotide encoding a polypeptide having phosphotransacetylase activity. Such cells exhibit a reduced or eliminated requirement for exogenous two-carbon substrate supplementation for their growth compared to PDC-KO cells. Applicants have also provided methods of making and using such recombinant host cells including, for example, methods of increasing cell growth, methods of reducing or eliminating the requirement of an exogenous two-carbon substrate for cell growth, methods of increasing glucose consumption and methods of increasing the production of a product of a pyruvate-utilizing pathway.
(50) Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In case of conflict, the present application including the definitions will control. Unless otherwise required, by context, singular terms shall include pluralities and plural terms shall include the, singular. All publications, patents and other references mentioned herein are incorporated by reference in their entireties for all purposes as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference, unless only specific sections of patents or patent publications are indicated to be incorporated by reference.
(51) Although methods and materials similar or equivalent to those described herein can be used in practice or testing of the present invention, suitable methods and materials are described below. The materials, methods and examples are illustrative only and are not intended to be limiting. Other features and advantages of the invention will be apparent from the detailed description and from the claims.
(52) In order to further define this invention, the following terms, abbreviations and definitions are provided.
(53) As used herein, the terms comprises, comprising, includes, including, has, having, contains, or containing, or any other variation thereof, are intended to be non-exclusive or open-ended. For example, a composition, a mixture, a process, a method, an article, or an apparatus that comprises a list of elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, mixture, process, method, article, or apparatus. Further, unless expressly stated to the contrary, or refers to an inclusive or and not to an exclusive or. For example, a condition A or B is satisfied by any one of the following: A is true (or present) and B is false (or not present), A is false (or not present) and B is true (or present), and both A and B are true (or present).
(54) Also, the indefinite articles a and an preceding an element or component of the invention are intended to be nonrestrictive regarding the number of instances, i.e., occurrences of the element or component. Therefore a or an should be read to include one or at least one, and the singular word form of the element or component also includes the plural unless the number is obviously meant to be singular.
(55) The term invention or present invention as used herein is a non-limiting term and is not intended to refer to any single embodiment of the particular invention but encompasses all possible embodiments as described in the application.
(56) As used herein, the term about modifying the quantity of an ingredient or reactant of the invention employed refers to variation in the numerical quantity that can occur, for example, through typical measuring and liquid handling procedures used for making concentrates or use, solutions in the real world; through inadvertent error in these procedures; through differences in the manufacture, source, or purity of the ingredients employed to make the compositions or to carry out the methods; and the like. The term about also encompasses amounts that differ due to different equilibrium conditions for a composition resulting from a particular initial mixture. Whether or not modified by the term about, the claims include equivalents to the quantities. In one embodiment, the term about means within 10% of the reported numerical value, preferably within 5% of the reported numerical value.
(57) The term butanol as used herein, refers to 2-butanol, 1-butanol, isobutanol, or mixtures thereof.
(58) The term pyruvate-utilizing biosynthetic pathway refers to an enzyme pathway to produce a biosynthetic product from pyruvate.
(59) The term isobutanol biosynthetic pathway refers to an enzyme pathway to produce isobutanol from pyruvate.
(60) The term 2-butanol biosynthetic pathway refers to an enzyme pathway to produce 2-butanol from pyruvate.
(61) The term 2-butanone biosynthetic pathway refers to an enzyme pathway to produce 2-butanone from pyruvate.
(62) The terms pdc-, PDC knock-out, or PDC-KO as used herein refer to a cell that has a genetic modification to inactivate or reduce expression of at least one gene encoding pyruvate decarboxylase (PDC) so that the cell substantially or completely lacks pyruvate decarboxylase enzyme activity. If the cell has more than one expressed (active) PDC gene, then each of the active PDC genes may be inactivated or have minimal expression thereby producing a pdc-cell.
(63) The term carbon substrate refers to a carbon source capable of being metabolized by the recombinant host cells disclosed herein. Non-limiting examples of carbon substrates are provided herein and include, but are not limited to, monosaccharides, oligosaccharides, polysaccharides, ethanol, lactate, succinate, glycerol, carbon dioxide, methanol, glucose, fructose, sucrose, xylose, arabinose, dextrose, or mixtures thereof.
(64) The term exogenous two-carbon substrate refers to the carbon source provided to be metabolized into acetyl-CoA by a host cell that lacks the ability to convert pyruvic acid into acetyl-CoA. The term is used to distinguish from the carbon substrate which is converted into a pyruvate-derived product by a pyruvate-utilizing biosynthetic pathway, herein also referred to as the pathway substrate which includes, for example, glucose.
(65) The term polynucleotide is intended to encompass a singular nucleic acid as well as plural nucleic acids, and refers to a nucleic acid molecule or construct, e.g., messenger RNA (mRNA) or plasmid DNA (pDNA). A polynucleotide can contain the nucleotide sequence of the full-length cDNA sequence, or a fragment thereof, including the untranslated 5 and 3 sequences and the coding sequences. The polynucleotide can be composed of any polyribonucleotide or polydeoxyribonucleotide, which may be unmodified RNA or DNA or modified RNA or DNA. For example, polynucleotides can be composed of single- and double-stranded DNA, DNA that is a mixture of single- and double-stranded regions, single- and double-stranded RNA, and RNA that is mixture of single- and double-stranded regions, hybrid molecules comprising DNA and RNA that may be single-stranded or, more typically, double-stranded or a mixture of single- and double-stranded regions. Polynucleotide embraces chemically, enzymatically, or metabolically modified forms.
(66) A polynucleotide sequence may be referred to as isolated, in which it has been removed from its native environment. For example, a heterologous polynucleotide encoding a polypeptide or polypeptide fragment having dihydroxy-acid dehydratase activity contained in a vector is considered isolated for the purposes of the present invention. Further examples of an isolated polynucleotide include recombinant polynucleotides maintained in heterologous host cells or purified (partially or substantially) polynucleotides in solution. Isolated polynucleotides or nucleic acids according to the present invention further include such molecules produced synthetically. An isolated polynucleotide fragment in the form of a polymer of DNA may be comprised of one or more segments of cDNA, genomic DNA or synthetic DNA.
(67) The term gene refers to a nucleic acid fragment that is capable of being expressed as a specific protein, optionally including regulatory sequences preceding (5 non-coding sequences) and following (3 non-coding sequences) the coding sequence. Native gene refers to a gene as found in nature with its own regulatory sequences. Endogenous gene refers to a native gene in its natural location in the genome of an organism. A heterologous gene refers to a gene not normally found in the host organism, but that is introduced into the host organism by gene transfer. Heterologous gene includes a native coding region, or portion thereof, that is reintroduced into the source organism in a form that is different from the corresponding native gene. For example, a heterologous gene may include a native coding region that is a portion of a chimeric gene including non-native regulatory regions that is reintroduced into the native host. Foreign genes can comprise native genes inserted into a non-native organism, or chimeric genes.
(68) As used herein the term coding region refers to a DNA sequence that codes for a specific amino acid sequence. Suitable regulatory sequences refer to nucleotide sequences located upstream (5 non-coding sequences), within, or downstream (3 non-coding sequences) of a coding sequence, and which influence the transcription, RNA processing or stability, or translation of the associated coding sequence. Regulatory sequences may include promoters, translation leader sequences, introns, polyadenylation recognition sequences, RNA processing site, effector binding site and stem-loop structure.
(69) As used herein, the term polypeptide is intended to encompass a singular polypeptide as well as plural polypeptides, and refers to a molecule composed of monomers (amino acids) linearly linked by amide bonds (also known as peptide bonds). The term polypeptide refers to any chain or chains of two or more amino acids, and does not refer to a specific length of the product. Thus, peptides, dipeptides, tripeptides, oligopeptides, protein, amino acid chain, or any other term used to refer to a chain or chains of two or more amino acids, are included within the definition of polypeptide, and the term polypeptide may be used instead of, or interchangeably with any of these terms. A polypeptide may be derived from a natural biological source or produced by recombinant technology, but is not necessarily translated from a designated nucleic acid sequence. It may be generated in any manner, including by chemical synthesis.
(70) By an isolated polypeptide or a fragment, variant, or derivative thereof is intended a polypeptide that is not in its natural milieu. No particular level of purification is required. For example, an isolated polypeptide can be removed from its native or natural environment. Recombinantly produced polypeptides and proteins expressed in host cells are considered isolated for purposed of the invention, as are native or recombinant polypeptides which have been separated, fractionated, or partially or substantially purified by any suitable technique.
(71) As used herein, the term valiant refers to a polypeptide differing from a specifically recited polypeptide of the invention by amino acid insertions, deletions, mutations, and substitutions, created using, e.g., recombinant DNA techniques, such as mutagenesis. Guidance in determining which amino acid residues may be replaced, added, or deleted without abolishing activities of interest, may be found by comparing the sequence of the particular polypeptide with that of homologous polypeptides, e.g., yeast or bacterial, and minimizing the number of amino acid sequence changes made in regions of high homology (conserved regions) or by replacing amino acids with consensus sequences.
(72) Alternatively, recombinant polynucleotide variants encoding these same or similar polypeptides may be synthesized or selected by making use of the redundancy in the genetic code. Various codon substitutions, such as silent changes which produce various restriction sites, may be introduced to optimize cloning into a plasmid or viral vector for expression. Mutations in the polynucleotide sequence may be reflected in the polypeptide or domains of other peptides added to the polypeptide to modify the properties of any part of the polypeptide.
(73) Amino acid substitutions may be the result of replacing one amino acid with another amino acid having similar structural and/or chemical properties, i.e., conservative amino acid replacements, or they may be the result of replacing, one amino acid with an amino acid having different structural and/or chemical properties, i.e., non-conservative amino acid replacements. Conservative amino acid substitutions may be made on the basis of similarity in polarity, charge, solubility, hydrophobicity, hydrophilicity, or the amphipathic nature of the residues involved. For example, nonpolar (hydrophobic) amino acids include alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan, and methionine; polar neutral amino acids include glycine, serine, threonine, cysteine, tyrosine, asparagine, and glutamine; positively charged (basic) amino acids include arginine, lysine, and, histidine; and negatively charged (acidic) amino acids include aspartic acid and glutamic acid. Alternatively, non-conservative amino acid substitutions may be made by selecting the differences in polarity, charge, solubility, hydrophobicity, hydrophilicity, or the amphipathic nature of any of these amino acids. Insertions or deletions may be within the range of variation as structurally or functionally tolerated by the recombinant proteins. The variation allowed may be experimentally determined by systematically making insertions, deletions, or substitutions of amino acids in a polypeptide molecule using recombinant DNA techniques and assaying the resulting recombinant variants for activity.
(74) The term promoter refers to a DNA sequence capable of controlling the expression of a coding sequence or functional RNA. In general, a coding sequence is located 3 to a promoter sequence. Promoters may be derived in their entirety from a native gene, or be composed of different elements derived from different promoters found in nature, or even comprise synthetic DNA segments. It is understood by those skilled in the art that different promoters may direct the expression of a gene in different tissues or cell types, or at different stages of development, or in response to different environmental or physiological conditions. Promoters which cause a gene to be expressed in most cell types at most times are commonly referred to as constitutive promoters. It is further recognized that since in most cases the exact boundaries of regulatory sequences have not been completely defined, DNA fragments of different lengths may have identical promoter activity. For example, it will be understood that FBA1 promoter can be used to refer to a fragment derived from the promoter region of the FBA1 gene.
(75) The term terminator as used herein refers to DNA sequences located downstream of a coding sequence. This includes polyadenylation recognition sequences and other sequences encoding regulatory signals capable of affecting mRNA processing or gene expression. The polyadenylation signal is usually characterized by affecting the addition of polyadenylic acid tracts to the 3 end of the mRNA precursor. The 3 region can influence the transcription, RNA processing or stability, or translation of the associated coding sequence. It is recognized that since in most cases the exact boundaries of regulatory sequences have not been completely defined, DNA fragments of different lengths may have identical terminator activity. For example, it will be understood that CYC1 terminator can be used to refer to a fragment derived from the terminator region of the CYC1 gene.
(76) The term operably linked refers to the association of nucleic acid sequences on a single nucleic acid fragment so that the function of one is affected by the other. For example, a promoter is operably linked with a coding sequence when it is capable of effecting the expression of that coding sequence (i.e., that the coding sequence is under the transcriptional control of the promoter). Coding sequences can be operably linked to regulatory sequences in sense or antisense orientation.
(77) The term expression, as used herein, refers to the transcription and stable accumulation of sense (mRNA) or antisense RNA derived from the nucleic acid fragment of the invention. Expression may also refer to translation of mRNA into a polypeptide.
(78) The term overexpression, as used herein, refers to expression that is higher than endogenous expression of the same or related gene. A heterologous gene is overexpressed if its expression is higher than that of a comparable endogenous gene.
(79) As used herein the term transformation refers to the transfer of a nucleic acid fragment into a host organism, resulting in genetically stable inheritance. Host organisms containing the transformed nucleic acid fragments are referred to as transgenic or recombinant or transformed organisms.
(80) The terms plasmid and vector as used herein, refer to an extra chromosomal element often carrying genes which are not part of the central metabolism of the cell, and usually in the form of circular double-stranded DNA molecules. Such elements may be autonomously replicating sequences, genome integrating sequences, phage or nucleotide sequences, linear or circular, of a single- or double-stranded DNA or RNA, derived from any source, in which a number of nucleotide sequences have been joined or recombined into a unique construction which is capable of introducing a promoter fragment and DNA sequence for a selected gene product along with appropriate 3 untranslated sequence into a cell.
(81) As used herein the term codon degeneracy refers to the nature in the genetic code permitting variation of the nucleotide sequence without affecting the amino acid sequence of an encoded polypeptide. The skilled artisan is well aware of the codon-bias exhibited by a specific host cell in usage of nucleotide codons to specify a given amino acid. Therefore, when synthesizing a gene for improved expression in a host cell, it is desirable to design the gene such that its frequency of codon usage approaches the frequency of preferred codon usage of the host cell.
(82) The term codon-optimized as it refers to genes or coding regions of nucleic acid molecules for transformation of various hosts, refers to the alteration of codons in the gene or coding regions of the nucleic acid molecules to reflect the typical codon usage of the host organism without altering the polypeptide encoded by the DNA. Such optimization includes replacing at least one, or more than one, or a significant number, of codons with one or more codons that are more frequently used in the genes of that organism.
(83) Deviations in the nucleotide sequence that comprise the codons encoding the amino acids of any polypeptide chain allow for variations in the sequence coding for the gene. Since each codon consists of three nucleotides, and the nucleotides comprising DNA are restricted to four specific bases, there are 64 possible combinations of nucleotides, 61 of which encode amino acids (the remaining three codons encode signals ending translation). The genetic code which shows which codons encode which amino acids is reproduced herein as Table 1. As a result, many amino acids are designated by more than one codon. For example, the amino acids alanine and proline are coded for by four triplets, serine and arginine by six, whereas tryptophan and methionine are coded by just one triplet. This degeneracy allows for DNA base composition to vary over a wide range without altering the amino acid sequence of the proteins encoded by the DNA.
(84) TABLE-US-00001 TABLE1 TheStandardGeneticCode T C A G T TTTPhe(F) TCTSer(S) TATTyr(Y) TGTCys(C) TTC TCC TAC TGC TTALeu(L) TCA TAAStop TGAStop TTG TCG TAGStop TGGTrp(W) C CTTLeu(L) CCTPro(P) CATHis(H) CGTArg(R) CTC CCC CAC CGC CTA CCA CAAGln(Q) CGA CTG CCG CAG CGG A ATTIle(I) ACTThr(T) AATAsn(N) AGTSer(S) ATC ACC AAC AGC ATA ACA AAALys(K) AGAArg(R) ATGMet(M) ACG AAG AGG G GTTVal(V) GCTAla(A) GATAsp(D) GGTGly(G) GTC GCC GAC GGC GTA GCA GAAGlu(E) GGA GTG GCG GAG GGG
(85) Many organisms display a bias for use of particular codons to code for insertion of a particular amino acid in a growing peptide chain. Codon preference, or codon bias, differences in codon usage between organisms, is afforded by degeneracy of the genetic code, and is well documented among many organisms. Codon bias often correlates with the efficiency of translation of messenger RNA (mRNA), which is in turn believed to be dependent on, inter alia, the properties of the codons being translated and the availability of particular transfer RNA (tRNA) molecules. The predominance of selected tRNAs in a cell is generally a reflection of the codons used most frequently in peptide synthesis. Accordingly, genes can be tailored for optimal gene expression in a given organism based on codon optimization.
(86) Given the large number of gene sequences available for a wide variety of animal, plant and microbial species, it is possible to calculate the relative frequencies of codon usage. Codon usage tables are readily available, for example, at the Codon Usage Database available at http://www.kazusa.or.jp/codon/ (visited Mar. 20, 2008), and these tables can be adapted in a number of ways. See Nakamura, Y., et al. Nucl. Acids Res. 28:292 (2000). Codon usage tables for yeast, calculated from GenBank Release 128.0 [15 Feb. 2002], are reproduced below as Table 2. This table uses mRNA nomenclature, and so instead of thymine (T) which is found in DNA, the tables use uracil (U) which is found in RNA. Table 2 has been adapted so that frequencies are calculated for each amino acid, rather than for all 64 codons.
(87) TABLE-US-00002 TABLE2 CodonUsageTableforSaccharomyces cerevisiaeGenes Frequencyper AminoAcid Codon Number thousand Phe UUU 170666 26.1 Phe UUC 120510 18.4 Leu UUA 170884 26.2 Leu UUG 177573 27.2 Leu CUU 80076 12.3 Leu CUC 35545 5.4 Leu CUA 87619 13.4 Leu CUG 68494 10.5 Ile AUU 196893 30.1 Ile AUC 112176 17.2 Ile AUA 116254 17.8 Met AUG 136805 20.9 Val GUU 144243 22.1 Val GUC 76947 11.8 Val GUA 76927 11.8 Val GUG 70337 10.8 Ser UCU 153557 23.5 Ser UCC 92923 14.2 Ser UCA 122028 18.7 Ser UCG 55951 8.6 Ser AGU 92466 14.2 Ser AGC 63726 9.8 Pro CCU 88263 13.5 Pro CCC 44309 6.8 Pro CCA 119641 18.3 Pro CCG 34597 5.3 Thr ACU 132522 20.3 Thr ACC 83207 12.7 Thr ACA 116084 17.8 Thr ACG 52045 8.0 Ala GCU 138358 21.2 Ala GCC 82357 12.6 Ala GCA 105910 16.2 Ala GCG 40358 6.2 Tyr UAU 122728 18.8 Tyr UAC 96596 14.8 His CAU 89007 13.6 His CAC 50785 7.8 Gln CAA 178251 27.3 Gln CAG 79121 12.1 Asn AAU 233124 35.7 Asn AAC 162199 24.8 Lys AAA 273618 41.9 Lys AAG 201361 30.8 Asp GAU 245641 37.6 Asp GAC 132048 20.2 Glu GAA 297944 45.6 Glu GAG 125717 19.2 Cys UGU 52903 8.1 Cys UGC 31095 4.8 Trp UGG 67789 10.4 Arg CGU 41791 6.4 Arg CGC 16993 2.6 Arg CGA 19562 3.0 Arg CGG 11351 1.7 Arg AGA 139081 21.3 Arg AGG 60289 9.2 Gly GGU 156109 23.9 Gly GGC 63903 9.8 Gly GGA 71216 10.9 Gly GGG 39359 6.0 Stop UAA 6913 1.1 Stop UAG 3312 0.5 Stop UGA 4447 0.7
(88) By utilizing this or similar tables, one of ordinary skill in the art can apply the frequencies to any given polypeptide sequence, and produce a nucleic acid fragment of a codon-optimized coding region which encodes the polypeptide, but which uses codons optimal for a given species.
(89) Randomly assigning codons at an optimized frequency to encode a given polypeptide sequence, can be done manually by calculating codon frequencies for each amino acid, and then assigning the codons to the polypeptide sequence randomly. Additionally, various algorithms and computer software programs are readily available to those of ordinary skill in the art. For example, the EditSeq function in the Lasergene Package, available from DNAstar, Inc., Madison, Wis., the backtransation function in the VectorNTI Suite, available from InforMax, Inc., Bethesda, Md., and the backtranslate function in the GCG-Wisconsin Package, available from Accelrys, Inc., San Diego, Calif. In addition, various resources are publicly available to codon-optimize coding region sequences, e.g., the backtranslation function at http://www.entelechon.com/bioinformatics/backtranslation.php?lang=eng (visited Apr. 15, 2008) and the backtranseq function available at http://bioinfo.pbi.nrc.ca:8090/EMBOSS/index.html (visited Jul. 9, 2002). Constructing a rudimentary algorithm to assign, codons based on a given frequency can also easily be accomplished with basic mathematical functions by one of ordinary skill in the art.
(90) Codon-optimized coding regions can be designed by various methods known to those skilled in the art including software packages such as synthetic gene designer (http://phenotype.biosci.umbc.edu/codon/sal/index.php).
(91) A polynucleotide or nucleic acid fragment is hybridizable to another nucleic acid fragment, such as a cDNA, genomic DNA, or RNA molecule, when a single-stranded form of the nucleic acid fragment can anneal to the other nucleic acid fragment under the appropriate conditions of temperature and solution ionic strength. Hybridization and washing conditions are well known and exemplified in Sambrook, J., Fritsch, E. F. and Maniatis, T. Molecular Cloning: A Laboraton Manual, 2.sup.nd ed., Cold Spring Harbor Laboratory: Cold Spring Harbor, N.Y. (1989), particularly Chapter 11 and Table 11.1 therein (entirely incorporated herein by reference). The conditions of temperature and ionic strength determine the stringency of the hybridization. Stringency conditions can be adjusted to screen for moderately similar fragments (such as homologous sequences from distantly related organisms), to highly similar fragments (such as genes that duplicate functional enzymes from closely related organisms). Post-hybridization washes determine stringency conditions. One set of preferred conditions uses a series of washes starting with 6SSC, 0.5% SDS at room temperature for 15 min, then repeated with 2SSC, 0.5% SDS at 45 C. for 30 min, and then repeated twice with 0.2SSC, 0.5% SDS at 50 C. for 30 min. A more preferred set of stringent conditions uses higher temperatures in which the washes are identical to those above except for the temperature of the final two 30 min washes in 0.2SSC, 0.5% SDS was increased to 60 C. Another preferred set of highly stringent conditions uses two final washes in 0.1SSC, 0.1% SDS at 65 C. An additional set of stringent conditions include hybridization at 0.1SSC, 0.1% SDS, 65 C. and washes with 2SSC, 0.1% SDS followed by 0.1SSC, 0.1% SDS, for example.
(92) Hybridization requires that the two nucleic acids contain complementary sequences, although depending on the stringency of the hybridization, mismatches between bases are possible. The appropriate stringency for hybridizing nucleic acids depends on the length of the nucleic acids and the degree of complementation, variables well known in the art. The greater the degree of similarity or homology between two nucleotide sequences, the greater the value of Tm for hybrids of nucleic acids having those sequences. The relative stability (corresponding to higher Tin) of nucleic acid hybridizations decreases in the following order: RNA:RNA, DNA:RNA, DNA:DNA. For hybrids of greater than 100 nucleotides in length, equations for calculating Tm have been derived (see Sambrook et al., supra, 9.50-9.51). For hybridizations with shorter nucleic acids, i.e., oligonucleotides, the position of mismatches becomes more important, and the length of the oligonucleotide determines its specificity (see Sambrook et al., supra, 11.7-11.8). In one embodiment the length for a hybridizable nucleic acid is at least about 10 nucleotides. Preferably a minimum length for a hybridizable nucleic acid is at least about 15 nucleotides; more preferably at least about 20 nucleotides; and most preferably the length is at least about 30 nucleotides. Furthermore, the skilled artisan will recognize that the temperature and wash solution salt concentration may be adjusted as necessary according to factors such as length of the probe.
(93) A substantial portion of an amino acid or nucleotide sequence is that portion comprising enough of the amino acid sequence, of a polypeptide or the nucleotide sequence of a gene to putatively identify that polypeptide or gene, either by manual evaluation of the sequence by one skilled in the art, or by computer-automated sequence comparison and identification using algorithms such as BLAST (Altschul, S. F., et al., J. Mol. Biol., 215:403-410 (199.3)). In general, a sequence often or more contiguous amino acids or thirty or more nucleotides is necessary in order to putatively identify a polypeptide or nucleic acid sequence as homologous to a known protein or gene. Moreover, with respect to nucleotide sequences, gene specific oligonucleotide probes comprising 20-30 contiguous nucleotides may be used in sequence-dependent methods of gene identification (e.g., Southern hybridization) and isolation (e.g., in situ hybridization of bacterial colonies or bacteriophage plaques). In addition, short oligonucleotides of 12-15 bases may be used as amplification primers in PCR in order to obtain a particular nucleic acid fragment comprising the primers. Accordingly, a substantial portion of a nucleotide sequence comprises enough of the sequence to specifically identify and/or isolate a nucleic acid fragment comprising the sequence. The instant specification teaches the complete amino acid and nucleotide sequence encoding particular proteins. The skilled artisan, having the benefit of the sequences as reported herein, may now use all or a substantial portion of the disclosed sequences for purposes known to those skilled in this art. Accordingly, the instant invention comprises the complete sequences as reported in the accompanying Sequence Listing, as well as substantial portions of those sequences as defined above.
(94) The team complementary is used to describe the relationship between nucleotide bases that are capable of hybridizing to one another. For example, with respect to DNA, adenosine is complementary to thymine and cytosine is complementary to guanine.
(95) The term percent identity, as known in the art, is a relationship between two or more polypeptide sequences or two or more polynucleotide sequences, as determined by comparing the sequences. In the art, identity also means the degree of sequence relatedness between polypeptide or polynucleotide sequences, as the case may be, as determined by the match between strings of such sequences. Identity and similarity can be readily calculated by known methods, including but not limited to those described in 1.) Computational Molecular Biology (Lesk, A. M., Ed.) Oxford University: NY (1988); 2.) Biocomputing: Informatics and Genome Projects (Smith, D. W., Ed.) Academic: NY (1993); 3.) Computer Analysis of Sequence Data Part I (Griffin, A. M., and Griffin, H. G., Eds.) Humania: NJ (1994); 4.) Sequence Analysis in Molecular Biology (von Heinje, G., Ed.) Academic (1987); and 5.) Sequence Analysis Primer (Gribskov, M. and Devereux, J., Eds.) Stockton: NY (1991).
(96) Preferred methods to determine identity are designed to give the best match between the sequences tested. Methods to determine identity and similarity are codified in publicly available computer programs. Sequence alignments and percent identity calculations may be performed using the MegAlign program of the LASERGENE bioinformatics computing suite (DNASTAR Inc., Madison, Wis.). Multiple alignment of the sequences is performed using the Clustal method of alignment which encompasses several varieties of the algorithm including the Clustal V method of alignment corresponding to the alignment method labeled Clustal V (described by Higgins and Sharp, CABIOS. 5:151-153 (1989); Higgins, D. G. et al., Comput. Appl. Biosci., 8:189-191 (1992)) and found in the MegAlign program of the LASERGENE bioinformatics computing suite (DNASTAR Inc.). For multiple alignments, the default values correspond to GAP PENALTY=10 and GAP LENGTH PENALTY-10. Default parameters for pairwise alignments and calculation of percent identity of protein sequences using the Clustal method are KTUPLE=1, GAP PENALTY-3, WINDOW-5 and DIAGONALS SAVED=5. For nucleic acids these parameters are KTUPLE=2, GAP PENALTY=5, WINDOW=4 and DIAGONAS SAVED=4. After alignment of the sequences using the Clustal V program, it is possible to obtain a percent identity by viewing the sequence distances table in the same program. Additionally the Clustal W method of alignment is available and corresponds to the alignment method labeled Clustal W (described by Higgins and Sharp, CABIOS. 5:151-153 (1989); Higgins, D. G. et al., Comput. Appl. Biosci. 8:189-191 (1992)) and found in the MegAlign v6.1 program of the LASERGENE bioinformatics computing suite (DNASTAR Inc.). Default parameters for multiple alignment (GAP PENALTY=10, GAP LENGTH PENALTY=0.2, Delay Divergen Seqs(%)=30, DNA Transition Weight=0.5, Protein Weight Matrix=Gonnet Series, DNA Weight Matrix=IUB). After alignment of the sequences using the Clustal W program, it is possible to obtain a percent identity by viewing the sequence distances table in the same program.
(97) It is well understood by one skilled in the art that many levels of sequence identity are useful in identifying polypeptides, such as from other species, wherein such polypeptides have the same or similar function or activity. Useful examples of percent identities include, but are not limited to: 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%, or any integer percentage from 55% to 100% may be useful in describing the present invention, such as 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%. Suitable nucleic acid fragments not only have the above homologies but typically encode a polypeptide having at least 50 amino acids, preferably at least 100 amino acids, more preferably at least 150 amino acids, still more preferably at least 200 amino acids, and most preferably at least 250 amino acids.
(98) The term sequence analysis software refers to any computer algorithm or software program that is useful for the analysis of nucleotide or amino acid sequences. Sequence analysis software may be commercially available or independently developed. Typical sequence analysis software will include, but is not limited to: 1.) the GCG suite of programs (Wisconsin Package Version 9.0, Genetics Computer Group (GCG), Madison, Wis.); 2.) BLASTP, BLASTN, BLASTX (Altschul et al., J. Mol. Biol., 215:403-410 (1990)); 3.) DNASTAR (DNASTAR, Inc. Madison, Wis.); 4.) Sequencher (Gene Codes Corporation, Ann Arbor, Mich.); and 5.) the FASTA program incorporating the Smith-Waterman algorithm (W. R. Pearson, Comput. Methods Genome Res., [Proc. Int. Symp.] (1994), Meeting Date 1992, 111-20. Editor(s): Suhai, Samlor. Plenum: New York, N.Y.). Within the context of this application it will be understood that where sequence analysis software is used for analysis, that the results of the analysis will be based on the default values of the program referenced, unless otherwise specified. As used herein default values will mean any set of values or parameters that originally load with the software when first initialized.
(99) Standard recombinant DNA and molecular cloning techniques used here are well known in the art and are described by Sambrook, J., Fritsch, E. F. and Maniatis, T., Molecular Cloning: A Laboratory Manual, Second Edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1989) (hereinafter Maniatis); and by Silhavy, T. J., Bennan, M. L. and Enquist, L. W., Experiments with Gene Fusions, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1984); and by Ausubel, F. M, et al., Current Protocols in Molecular Biology, published by Greene Publishing Assoc. and Wiley-Interscience (1987). Additional methods used here are in Methods in Enzymology, Volume 194, Guide to Yeast Genetics and Molecular and Cell Biology (Part A, 2004, Christine Guthrie and Gerald R. Fink (Eds.), Elsevier Academic Press, San Diego, Calif.). Other molecular tools and techniques are known in the art and include splicing by overlapping extension polymerase chain reaction (PCR) (Yu, et al. (2004) Fungal Genet. Biol. 41:973-981), positive selection for mutations at the URA3 locus of Saccharomyces cerevisiae (Boeke, J. D. et al. (1984) Mol. Gen. Genet. 197, 345-346; M A Romanos, et al. Nucleic Acids Res. 1991 Jan. 11; 19(1): 187), the cre-lox site-specific recombination system as well as mutant lox sites and FLP substrate mutations (Sauer, B. (1987) Mol Cell Biol 7: 2087-2096; Senecoff, et al. (1988) Journal of Molecular Biology, Volume 201, Issue 2, Pages 405-421; Albert, et al, (1995) The Plant Journal. Volume 7, Issue 4, pages 649-659), seamless gene deletion (Akada, et al. (2006) Yeast; 23(5):399-405), and gap repair methodology (Ma et al., Genetics 58:201-216; i. 981). Applicants have discovered that activation of the phosphoketolase pathway in a recombinant host cell comprising a modification in an endogenous gene encoding a polypeptide having pyruvate decarboxylase activity or a modification in an endogenous polypeptide having pyruvate decarboxylase activity, reduces or eliminates the need for an exogenous carbon substrate for the growth of such a cell. In embodiments, the recombinant host cells comprise (i) at least one deletion, mutation and/or substitution in an endogenous gene encoding a polypeptide having pyruvate decarboxylase activity); (ii) a heterologous polynucleotide encoding a polypeptide having phosphoketolase activity; and optionally (iii) a heterologous polynucleotide encoding a polypeptide having phosphotransacetylase activity.
(100) The genetic manipulations of the host cells described herein can be performed using standard genetic techniques and screening and can be made in any host cell that is suitable to genetic manipulation (Methods in Yeast Genetic's, 2005, Cold Spring Harbor Laboratory Press Cold Spring Harbor, N.Y., pp. 201-202). In embodiments, the recombinant host cells disclosed herein can be any bacteria, yeast or fungi host useful for genetic modification and recombinant gene expression. In other embodiments a recombinant host cell can be a member of the genera Clostridium, Zymomonas, Eseherichia, Salmonella, Serratia, Erwinia, Klebsiella, Shigella, Rhodococcus, Pseudomonas, Bacillus, Lactobacillus, Enterococcus, Alcaligenes, Paenibacillus, Arthrobacter, Corynebacterium, Brevibacterium, Schizoaccharomyces, Kluyveromyces, Yarrowia, Pichia, Candida, Hansenula, Issatchenkia, saccharomyces. In other embodiments, the host cell can be Saccharomyces cerevisiae, Schizosaccharomyces pombe, Kluyveromyces lactic, Kluyveromyces thermotolerans, Kluyveromyces marxianus, Candida glabrata, Candida albicans, Pichia stipitis, Yarrowia lipolytica, E. coli, or L. plantarum. In still other embodiments, the host cell is a yeast host cell. In some embodiments, the host cell is a member of the genera Saccharomyces. In some embodiments, the host cell is Kluyveromyces Candida glabrata or Schizosaccharomyces pombe. In some embodiments, the host cell is Saccharomyces cerevisiae. S. cerevisiae yeast are known in the art and are available from a variety of sources, including, but not limited to, American Type Culture Collection (Rockville, Md.), Centraalbureau voor Schimmelcultures (CBS) Fungal Biodiversity Centre, LeSaffre, Gert Strand AB, Ferm Solutions, North American Bioproducts, Martrex, and Lallemand. S. cerevisiae include, but are not limited to, BY4741, CEN.PK 113-7D, Ethanol Red yeast, Ferm Pro yeast, Bio-Ferm XR yeast, Gert Strand Prestige Batch Turbo alcohol yeast, Gert Strand Pot Distillers yeast, Gert Strand Distillers Turbo yeast, FerMax Green yeast, FerMax Gold yeast, Thermosacc yeast, BG-1, PE-2, CAT-1, CBS7959, CBS7960, and CBS7961.
(101) Sources of Acetyl-CoA
(102) Acetyl-CoA is a major cellular building block, required for the synthesis of fatty acids, sterols, and lysine. Pyruvate is often a major contributor to the acetyl-CoA pool. Pyruvate dehydrogenase catalyzes the direct conversion of pyruvate to acetyl-CoA (E.C. 1.2.4.1, E.C. 1.2.1.51) or acetate (E.C. 1.2.2.2) and is almost ubiquitous in nature. Other enzymes involved in conversion of pyruvate to acetyl-CoA, acetyl-phosphate or acetate include pyruvate-formate lyase (E.C. 2.3.1.54), pyruvate oxidase (E.C. 1.2.3.3, E.C. 1.2.3.6), pyruvate-ferredoxin oxidoreductase (E.C. 1.2.7.1), and pyruvate decarboxylase (E.C. 4.1.1.1). Genetic modifications made to a host cell to conserve the pyruvate pool for a product of interest may include those that restrict conversion to acetyl-CoA, leading to decreased growth in the absence of an exogenously supplied two-carbon substrate, a carbon substrate that can be readily converted to acetyl-CoA independent of pyruvate (e.g. ethanol or acetate). An example is the documented auxotrophy observed in pyruvate decarboxylase deficient Saccharomyces cerevisiae (Flikweert et al. 1999, supra). Another example is the documented auxotrophy observed in pyruvate dehydrogenase deficient Escherichia coli when grown aerobically on glucose (Langley and Guest, 1977, J. Gen. Microbial. 99:2630276).
(103) Modification of Pyruvate Decarboxylase
(104) In embodiments, the recombinant host cells disclosed herein comprise a modification in an endogenous polynucleotide encoding a polypeptide having pyruvate decarboxylase (PDC) or a modification in an endogenous polypeptide having PDC activity. In embodiments, the recombinant host cells disclosed herein can have a modification or disruption of one or more polynucleotides, genes or polypeptides encoding PDC. In embodiments, the recombinant host cell comprises at least one deletion, mutation, and/or substitution in one of more endogenous polynucleotides or genes encoding a polypeptide having PDC activity, or in one or more endogenous polypeptides having PDC activity. Such modifications, disruptions, deletions, mutations, and/or substitutions can result in PDC activity that is reduced or eliminated, resulting in a PDC knock-out (PDC-KO) phenotype.
(105) In embodiments, the endogenous pyruvate decarboxylase activity of the recombinant host cells disclosed herein converts pyruvate to acetaldehyde, which can then be converted to ethanol or to acetyl-CoA via acetate.
(106) In embodiments, the recombinant host cell is Kluyveromyces lactis containing one gene encoding pyruvate decarboxylase, Candida glabrata containing one gene encoding pyruvate decarboxylase, or Schizosaccharomyces pombe containing one gene encoding pyruvate decarboxylase.
(107) In other embodiments, the recombinant host cell is Saccharomyces cerevisiae containing three isozymes of pyruvate decarboxylase encoded by the pdc1, pdc5, and pdc6 genes, as well as a pyruvate decarboxylase regulatory gene, pdc2. In a non-limiting example in S. cerevisiae, the pdc1 and pdc5 genes, or all three genes, are disrupted. In another non-limiting example in S. cerevisiae, pyruvate decarboxylase activity may be reduced by disrupting the pdc2 regulatory gene. In, another non-limiting example in S. cerevisiae, polynucleotides or genes encoding pyruvate decarboxylase proteins such as those having about 70% to about 75%, about 75% to about 80%, about 80% to about 85%, about 85% to about 90%, about 90% to about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity to pdc1 or pdc5 can be disrupted.
(108) In embodiments, the polypeptide having PDC activity or the polynucleotide or gene encoding a polypeptide having PDC activity is associated with Enzyme Commission Number EC 4.1.1.1. In other embodiments, a PDC gene of the recombinant host cells disclosed herein is not active under the fermentation conditions used, and therefore such a gene would not need to be modified or inactivated.
(109) Examples of recombinant host cells with reduced pyruvate decarboxylase activity due to disruption of pyruvate decarboxylase encoding genes have been reported, such as for Saccharomyces in Flikweert et al. (Yeast (1996) 12:247-257), for Kluyveromyces in Bianchi et al. (Mol. Microbiol. (1996) 19(1):27-36), and disruption of the regulatory gene in Hohmann (Mol. Gen. Genet. (1993) 241:657-666). Saccharomyces strains having no pyruvate decarboxylase activity are available from the ATCC with Accession #200027 and #200028.
(110) Examples of PDC polynucleotides, genes and polypeptides that can be targeted for modification or inactivation in the recombinant host cells disclosed herein include, but are not limited to, those of the following table.
(111) TABLE-US-00003 TABLE 3 SEQ ID NOs of pyruvate decarboxylase (PDC) target gene coding regions and proteins. SEQ ID NO: SEQ ID NO: Description Nucleic acid Amino acid PDC1 pyruvate decarboxylase 1 2 from Saccharomyces cerevisiae PDC5 pyruvate decarboxylase 3 4 from Saccharomyces cerevisiae PDC6 pyruvate decarboxylase 5 6 from Saccharomyces cerevisiae pyruvate decarboxylase from 7 8 Candida glabrata PDC1 pyruvate decarboxylase 9 10 from Pichia stipitis PDC2 pyruvate decarboxylase 11 12 from Pichia stipitis pyruvate decarboxylase from 13 14 Kluyveromyces lactis pyruvate decarboxylase from 15 16 Yarrowia lipolytica pyruvate decarboxylase from 17 18 Schizosaccharomyces pombe pyruvate decarboxylase from 18 20 Zygosaccharomyces rouxii
(112) Other examples of PDC polynucleotides, genes and polypeptides that can be targeted for modification or inactivation in the recombinant host cells disclosed herein include, but are not limited to, PDC polynucleotides, genes and/or polypeptides having at least about 70% to about 75%, about 75% to about 80%, about 80% to about 85%, about 85% to about 90%, about 90% to about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity to any one of the sequences of Table 3.
(113) In embodiments, the sequences of other PDC polynucleotides, genes and/or polypeptides can be identified in the literature and in bioinformatics databases well known to the skilled person using sequences disclosed herein and available in the art. For example, such sequences can be identified through BLAST (as described above) searching of publicly available databases with known PDC encoding polynucleotide or polypeptide sequences. In such a method, identities can be based on the Clustal W method of alignment using the default parameters of GAP PENALTY=10, GAP LENGTH PENALTY=0.1, and Gonnet 250 series of protein weight matrix.
(114) Additionally, the PDC polynucleotide or polypeptide sequences described herein or known the art can be used to identify other PDC homologs in nature. For example, each of the PDC encoding nucleic acid fragments described herein can be used to isolate genes encoding homologous proteins. Isolation of homologous genes using sequence-dependent protocols is well known in the art. Examples of sequence-dependent protocols include, but are not limited to (1) methods of nucleic acid hybridization; (2) methods of DNA and RNA amplification, as exemplified by various uses of nucleic acid amplification technologies [e.g., polymerase chain reaction (PCR), Mullis et al., U.S. Pat. No. 4,683,202; ligase chain reaction (LCR), Tabor, S. et al., Proc. Acad. Sci. USA 82:1074 (1985); or strand displacement amplification (SDA), Walker et al., Proc. Natl. Acad. Sci. USA., 89:392 (1992)]; and (3) methods of library construction and screening by complementation.
(115) In embodiments, PDC polynucleotides, genes and/or polypeptides related to the recombinant host cells described herein can be modified or disrupted. Many methods for genetic modification and disruption of target genes to reduce or eliminate expression are known to one of ordinary skill in the art and can be used to create the recombinant host cells described herein. Modifications that can be used include, but are not limited to, deletion of the entire gene or a portion of the gene encoding a PDC protein, inserting a DNA fragment into the encoding gene (in either the promoter or coding region) so that the protein is not expressed or expressed at lower levels, introducing a mutation into the coding region which adds a stop codon or frame shift such that a functional protein is not expressed, and introducing one or more mutations into the coding region to alter amino acids so that a non-functional or a less active protein is expressed. In other embodiments, expression of a target gene can be blocked by expression of an antisense RNA or an interfering RNA, and constructs can be introduced that result in, cosuppression. In other embodiments, the synthesis or stability of the transcript can be lessened by mutation. In embodiments, the efficiency by which a protein is translated from mRNA can be modulated by mutation. All of these methods can be readily practiced by one skilled in the art making use of the known or identified sequences encoding target proteins.
(116) In other embodiments, DNA sequences surrounding a target PDC coding sequence are also useful in some modification procedures and are available, for example, for yeasts such as Saccharomyces cerevisiae in the complete genome sequence coordinated by Genome Project ID9518 of Genome Projects coordinated by NCBI (National Center for Biotechnology Information) with identifying GOPID #13838. An additional non-limiting example of yeast genomic sequences is that of Candida albicans, which is included in GPID #10771, #10701 and #16373, Other yeast genomic sequences can be readily found by one of skill in the art in publicly available databases.
(117) In other embodiments, DNA sequences surrounding a target PDC coding sequence can be useful for modification methods using homologous recombination. In a non-limiting example of this method, PDC gene flanking sequences can be placed bounding a selectable marker gene to mediate homologous recombination whereby the marker gene replaces the PDC gene. In another non-limiting example, partial PDC gene sequences and PDC, gene flanking sequences bounding a selectable marker gene can be used to mediate homologous recombination whereby the marker gene replaces a portion of the target PDC, gene. In embodiments, the selectable marker can be bounded by site-specific recombination sites, so that following expression of the corresponding site-specific recombinase, the resistance gene is excised from the PDC gene without reactivating the latter. In embodiments, the site-specific recombination leaves behind a recombination site which disrupts expression of the PDC protein. In other embodiments, the homologous recombination vector can be constructed to also leave a deletion in the PDC gene following excision of the selectable marker, as is well known to one skilled in the art.
(118) In other embodiments, deletions can be made to a PDC target gene using mitotic recombination as described in Wach et al. (Yeast, 10:1793-1808; 1994). Such a method can involve preparing a DNA fragment that contains a selectable marker between genomic regions that can be as short as 20 bp, and which bound a target DNA sequence. In other embodiments, this DNA fragment can be prepared by PCR amplification of the selectable marker gene using as primers oligonucleotides that hybridize to the ends of the marker gene and that include the genomic regions that can recombine with the yeast genome. In embodiments, the linear DNA fragment can be efficiently transformed into yeast and recombined into the genome resulting in gene replacement including with deletion of the target DNA sequence ((as described, for example, in Methods in Enzymology, Volume 194, Guide to Yeast Genetics and Molecular and Cell Biology (Part A. 2004, Christine Guthrie and Gerald R. Fink (Eds.), Elsevier Academic Press, San Diego, Calif.)).
(119) Moreover, promoter replacement methods can be used to exchange the endogenous transcriptional control elements allowing another means to modulate expression such as described in Mnaimneh et al. ((2004) Cell 118(1):31-44).
(120) In other embodiments, the PDC target gene encoded activity can be disrupted using random mutagenesis, which can then be followed by screening to identify strains with dependency on carbon substrates for growth. In this type of method, the DNA sequence of the target gene encoding region, or any other region of the genome affecting carbon substrate dependency for growth, need not be known. In embodiments, a screen for cells with reduced PDC activity and/or two-carbon substrate dependency, or other mutants having reduced PDC activity and a reduced or eliminated dependency for exogenous two-carbon substrate for growth, can be useful as recombinant host cells of the invention.
(121) Methods for creating genetic mutations are common and well known in the art and can be applied to the exercise of creating mutants. Commonly used random genetic modification methods (reviewed in Methods in Yeast Genetics, 2005, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.) include spontaneous mutagenesis, mutagenesis caused by mutator genes, chemical mutagenesis, irradiation with UV or X-rays, or transposon mutagenesis.
(122) Chemical mutagenesis of host cells can involve, but is not limited to, treatment with one of the following DNA mutagens: ethyl methanesulfonate (EMS), nitrous acid, diethyl sulfate, or N-methyl-N-nitro-N-nitroso-guanidine (MNNG). Such methods of mutagenesis have been reviewed in Spencer et al., (Mutagenesis in Yeast, 1996, Yeast Protocols: Methods in Cell and Molecular Biology. Humana Press, Totowa, N.J.). In embodiments, chemical mutagenesis with EMS can be performed as described in Methods in Yeast Genetics, 2005, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. Irradiation with ultraviolet (UV) light or X-rays can also be used to produce random mutagenesis in yeast cells. The primary effect of mutagenesis by UV irradiation is the formation of pyrimidine dimers which disrupt the fidelity of DNA replication. Protocols for UV-mutagenesis of yeast can be found in Spencer et al. (Mutagenesis in Yeast, 1996, Yeast Protocols: Methods in Cell and Molecular Biology. Humana Press, Totowa, N.J.). In embodiments, the introduction of a mutator phenotype can also be used to generate random chromosomal mutations in host cells. In embodiments, common imitator phenotypes can be obtained through disruption of one or more of the following genes: PMS1, MAG1, RAD18 or RAD51. In other embodiments, restoration of the non-mutator phenotype can be obtained by insertion of the wildtype allele. In other embodiments, collections of modified cells produced from any of these or other known random mutagenesis processes may be screened for reduced or eliminated PDC activity.
(123) Genomes have been completely sequenced and annotated and are publicly available for the following yeast strains: Ashbya gossypii ATCC 10895, Candida glabrata CBS138, Kluyveromyces lactis NRRL Y-1140, Pichia stipitis CBS 6054, Saccharomyces cerevisiae S288c, Schizosaccharomyces pombe 972h-, and Yarrowia lipolytica CLIB 122. Typically BLAST (described above) searching of publicly available databases with known PDC polynucleotide or polypeptide sequences, such as those provided herein, is used to identify PDC-encoding sequences of other host cells, such as yeast cells.
(124) Accordingly, it is within the scope of the invention to provide pyruvate decarboxylase polynucleotides and polypeptides having at, least about 70% to about 75%, about 75% to about 80%, about 80% to about 85%, about 85% to about 90%, about 90% to about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity to any of the PDC polypeptides or polypeptides disclosed herein (SEQ ID NOs: 1-20). Identities are based on the Clustal W method of alignment using the default parameters of GAP PENALTY=10, GAP LENGTH PENALTY=0.1, and Gonnet 250 series of protein weight matrix.
(125) The modification of PDC in the host cells disclosed herein to reduce or eliminate PDC activity can be confirmed using methods known in the art. For example, PCR methods well known in the art can be used to confirm deletion of PDC. Other suitable methods will be known to those of skill in the art and include, but are not limited to lack of growth on yeast extract peptone-dextrose medium (YPD).
(126) Introduction of the Phosphokeetolase Pathway
(127) Applicants have found that expression of enzymes associated with the phosphoketolase pathway (e.g., phosphoketolase and/or phosphotransacetylase) results in a reduced or eliminated requirement for exogenous two-carbon substrate supplementation for growth of PDC-KO cells. Phosphoketolases and/or phosphotransacetylases identified as described herein, can be expressed in such cells using methods described herein.
(128) Enzymes of the phosphoketolase pathway include phosphoketolase and phosphotransacetylase (
(129) In embodiments, the phosphoketolase pathway is activated in the recombinant host cells disclosed herein by engineering the cells to express polynucleotides and/or polypeptides encoding phosphoketolase and, optionally, phosphotransacetylase. In embodiments, the recombinant host cells disclosed herein comprise a heterologous polynucleotide encoding a polypeptide having phosphoketolase activity. In other embodiments, the recombinant host cells disclosed herein comprise a heterologous polynucleotide encoding, a polypeptide having phosphoketolase activity and a heterologous polynucleotide encoding a polypeptide having phosphotransacetylase activity. In other embodiments, the heterologous polynucleotide encoding a polypeptide having phosphoketolase activity is overexpressed, or expressed at a level that is higher than endogenous expression of the same or related endogenous gene, if any. In still other embodiments, the heterologous polynucleotide encoding a polypeptide having phosphotransacetylase activity is overexpressed, or expressed at a level that is higher than endogenous expression of the same or related endogenous gene, if any.
(130) In embodiments, a polypeptide having phosphoketolase activity catalyzes the conversion of xylulose 5-phosphate into glyceraldehyde-3-phosphate and acetyl-phosphate and/or the conversion of fructose-6-phosphate into erythrose-4-phosphate and acetyl-phosphate. In embodiments, the activity of a polypeptide having phosphoketolase activity is inhibited by erythrose 4-phosphate and/or glyceraldehyde 3-phosphate. In other embodiments, a polypeptide having phosphotransacetylase activity catalyzes the conversion of acetyl-phosphate into acetyl-CoA.
(131) Numerous examples of polynucleotides, genes and polypeptides encoding phosphoketolase activity are known in the art and can be used in the recombinant host cells disclosed herein. In embodiments, such a polynucleotide, gene and/or polypeptide can be the xylulose 5-phosphateketolase (XpkA) of Lactobacillus pentosus MD363 (Posthuma et al., Appl. Environ Microbiol. 68: 831-7; 2002). XpkA is the central enzyme of the phosphoketolase pathway (PKP) in lactic acid bacteria, and exhibits a specific activity of 4.455 mol/min/mg (Posthuma et al., Appl. Environ. Microbiol. 68: 831-7; 2002). In other embodiments, such a polynucleotide, gene and/or polypeptide can be the phosphoketolase of Leuconostoc mesenteroides which exhibits a specific activity of 9.9 mol/min/mg and is stable at pH above 4.5 (Goldberg et al., Methods Enzymol. 9: 515-520; 1966). This phosphoketolase exhibits a Km of 4.7 mM for D-xylulose 5-phosphate and a Km of 29 mM for fructose 6-phosphate (Goldberg et al., Methods Enzymol. 9: 515-520; 1966). In other embodiments, such a polynucleotide, gene and/or polypeptide can be the D-xylulose 5-phosphate/D-fructose 6-phosphate phosphoketolase gene xfp from B. lactis, as described, for example, in a pentose-metabolizing S. cerevisiae strain by Sonderegger et al. (Appl. Environ. Microbiol. 70: 2892-7; 2004).
(132) In embodiments, a polynucleotide, gene and/or polypeptide encoding phosphoketolase corresponds to the Enzyme Commission Number EC 4.1.2.9.
(133) In embodiments, host cells comprise a polypeptide having at least about 80%, at least about 85%, at least about 90%, or 100% identity to a polypeptide of Table 4 or an, active fragment thereof or a polynucleotide encoding such a polypeptide. In other embodiments, a polynucleotide, gene and/or polypeptide encoding phosphoketolase can include, but is not limited to, a sequence provided in the following tables 4 or 5.
(134) TABLE-US-00004 TABLE4 SEQIDNOsofphosphoketolasetargetgenecodingregionsandproteins SEQID SEQID NO: NO: Nucleic Amino Description acid acid AminoAcidsequence Xpk1 172 481 MTTDYSSPAYLQKVDKYWRAANYLSVGQLYLKDNPLLQRPL phospho- KASDVKVHPIGHWGTIAGQ ketolasefrom NFIYAHLNRVINKYGLKMFYVEGPGHGGQVMVSNSYLDGTY Lactobacillus TDIYPEITQDVEGMQKLFK plantarum QFSFPGGVASHAAPETPGSIHEGGELGYSISHGVGAILDNP DEIAAVVVGDGESETGPLA TSWQSTKFINPINDGAVLPILNLNGFKISNPTIFGRTSDAK IKEYFESMNWEPIFVEGDD PEKVHPALAKAMDEAVEKIKAIQKHARENNDATLPVWPMIV FRAPKGWTGPKSWDGDKIE GSFRAHQIPIPVDQNDMEHADALVDWLESYQPKELFNEDGS LKDDIKEIIPTGDSRMAAN PITNGGVDPKALNLPNFRDYAVDTSKEGANVKQDMIVWSDY LRDVIKKNPDNFRLFGPDE TMSNRLYGVFETTNRQWMEDIHPDSDQYEAPAGRVLDAQLS EHQAEGWLEGYVLTGRHGL FASYEAFLRVVDSMLTQHFKWLRKANELDWRKKYPSLNIIA ASTVFQQDHNGYTHQDPGA LTHLAEKKPEYIREYLPADANTLLAVGDVIFRSQEKINYVV TSKHPRQQWFSIEEAKQLV DNGLGIIDWASTDQGSEPDIVFAAAGTEPTLETLAAIQLLH DSFPEMKIRFVNVVDILKL RSPEKDPRGLSDAEFDHYFTKDKPVVFAFHGYEDLVRDIFF DRHNHNLYVHGYRENGDIT TPFDVRVMNQMDRFDLAKSAIAAQPAMENTGAAFVQSMDNM LAKHNAYIRDAGTDLPEVN DWQWKGLK XpkA 1890 1889 MSTDYSSPAYLQKVDKYWRAANYLSVGQLYLKDNPLLQRPL phospho- KASDVKVHPIGHWGTIAGQ ketolasefrom NFIYAHLNRVINKYGLKMFYVEGPGHGGQVMVSNSYLDGTY Lactobacillus TDIYPEITQDVEGMQKLFK pentosus QFSFPGGVASHAAPETPGSIHEGGELGYSISHGVGAILDNP MD363 DEIAAVVVGDGESETGPLA TSWQSTKFINPINDGAVLPILNLNGFKISNPTIFGRTSDEK IKQYFESMNWEPIFVEGDD PEKVHPALAKAMDEAVEKIKAIQKNARENDDATLPVWPMIV FRAPKGWTGPKSWDGDKIE GSFRAHQIPIPVDQTDMEHADALVDWLESYQPKELFNEDGS LKDDIKEIIPTGDARMAAN PITNGGVDPKALNLPNFRDYAVDTSKHGANVKQDMIVWSDY LRDVIKKNPDNFRLFGPDE TMSNRLYGVFETTNRQWMEDIHPDSDQYEAPAGRVLDAQLS EHQAEGWLEGYVLTGRHGL FASYEAFLRVVDSMLTQHFKWLRKANELDWRKKYPSLNIIA ASTVFQQDHNGYTHQDPGA LTHLAEKKPEYIREYLPADANSLLAVGDVIFRSQEKINYVV TSKHPRQQWFSIEEAKQLV DNGLGIIDWASTDQGSEPDIVFAAAGTEPTLETLAAIQLLH DSFPDMKIRFVNVVDILKL RSPEKDPRGLSDAEFDHYFTKDKPVVFAFHGYEDLVRDIFF DRHNHNLHVHGYRENGDIT TPFDVRVMNQMDRFDLAKSAIAAQPAMENTGAAFVQDMDNM LAKHNAYIRDAGTDLPEVN DWQWKGLK XpfD-xylulose 79 388 MTNPVIGTPWQKLDRPVSEEAIEGMDKYWRVANYMSIGQIY 5-phosphate/D- LRSNPLMKEPFTRDDVKHR fructose6- LVGHWGTTPGLNFLLAHINRLIADHQQNTVFIMGPGHGGPA phosphate GTAQSYIDGTYTEYYPNIT phospho- KDEAGLQKFFRQFSYPGGIPSHFAPETPGSIHEGGELGYAL ketolase SHAYGAIMDNPSLFVPCII fromB.lactis GDGEAETGPLATGWQSNKLVNPRTDGIVLPILHLNGYKIAN PTILARISDEELHDFFRGM GYHPYEFVAGFDNEDHLSIHRRFAELFETIFDEICDIKAAA QTDDMTRPFYPMLIFRTPK GWTCPKFIDGKKTEGSWRAHQVPLASARDTEAHFEVLKGWM ESYKPEELFNADGSIKEDV TAFMPKGELRIGANPNANGGRIREDLKLPELDQYEITGVKE YGHGWGQVEAPRSLGAYCR DIIKNNPDSFRVFGPDETASNRLNATYEVTKKQWDNGYLSA LVDENMAVTGQVVEQLSEH QCEGFLEAYLLTGRHGIWSSYESFVHVIDSMLNQHAKWLEA TVREIPWRKPISSVNLLVS SHVWRQDHNGFSHQDPGVTSVLLNKTFNNDHVTNIYFATDA NMLLAIAEKCFKSTNKINA IFAGKQPAATWITLDEARAELEAGAAEWKWASNAKSNDEVQ VVLAAAGDVPTQEIMAASD ALNKMGIKFKVVNVVDLIKLQSSKENDEAMSDEDFADLFTA DKPVLFAYHSYAQDVRGLI YDRPNHDNFTVVGYKEQGSTTTPFDMVRVNDMDRYALQAKA LELIDADKYADKINELNEF RKTAFQFAVDNGYDIPEFTDWVYPDVKVDETSMLSATAATA GDNE
(135) In other embodiments, a polynucleotide, gene and/or polypeptide encoding phosphoketolase can have at least about 70% to about 75%, about 75% to about 80%, about 80% to about 85%, about 85% to about 90%, about 90% to about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity to that of any one of the sequences of Table 4, wherein the polynucleotide, gene and/or polypeptide encodes a polypeptide having phosphoketolase activity.
(136) In other embodiments, a polynucleotide, gene and/or polypeptide encoding phosphoketolase can be used to identify other phosphoketolase polynucleotide, gene and/or polypeptide sequences or to identify phosphoketolase homologs in other cells, as described above for PDC. Such phosphoketolase encoding sequences can be identified, for example, in, the literature and/or in bioinformatics databases well known to the skilled person. For example, the identification of phosphoketolase encoding sequences in other cell types using bioinformatics can be accomplished through BLAST (as described above) searching of publicly available databases with known phosphoketolase encoding DNA and polypeptide sequences, such as those provided herein. Identities are based on the Clustal W method of alignment using the default parameters of GAP PENALTY=10, GAP LENGTH PENALTY=0.1, and Gonnet 250 series of protein weight matrix.
(137) Additional phosphoketolase target gene coding regions were identified using, diversity search, clustering, experimentally verified xylulose-5-phosphate/fructose-6-phosphate phosphoketolases and domain architecture. Briefly, a BLAST search with the experimentally verified sequences with an Evalue cut-off of 0.01 resulted in 595 sequence matches. Clustering with the CD-HIT program at 95% sequence identity and 90% length overlap reduced the number to 436. CD-HIT is a program for clustering large protein database at nigh sequence identity threshold. The program removes redundant sequences and gel erates a database of only the representatives. (Clustering of highly homologous sequences to reduce the size of large protein database, Weizhong Li, Lukasz Jaroszewski & Adam Godzik Bioinformatics, (2001) 17:282-283)
(138) Xylulose-5-phosphate/fructose-6-phosphate phosphoketolases have three Pfam domains: XFP_N; XFP; XFP_C. Although each of these domains may be present in several domain architectures, e.g. XFP_N is found in eight architectures. The architecture of interest was determined to be XFP_N; XFP; XFP_C. The cumulative length of the three domains is 760 amino acids.
(139) A structure/function characterization of the phosphoketolases was performed using the HMMER software package. The following information based on the HMMER software user guide gives some description of the way that the hmmbuild program prepares a Profile HMM. A Profile HMM is capable of modeling gapped alignments, e.g. including insertions and deletions, which lets the software describe a complete conserved domain (rather than just a small ungapped motif). Insertions and deletions are modeled using insertion (I) states and deletion (D) states. All columns that contain more than a certain fraction x of gap characters will be assigned as an insert column. By default, x is set to 0.5. Each match state has an I and a D state associated with it. HMMER calls a group of three states (M/D/I) at the same consensus position in the alignment a node, These states are interconnected with arrows called state transition probabilities. M and I states are emitters, while D states are silent. The transitions are arranged so that at each node, either the M state is used (and a residue is aligned and scored) or the D state is, used (and no residue is aligned, resulting in a deletion-gap character, -). Insertions occur between nodes, and I states, have a self-transition, allowing one or more inserted residues to occur between consensus columns.
(140) The scores of residues in a match state (i.e. match state emission scores), or in an insert state (i.e. insert state emission scores) are proportional to Log_2 (p_x)/(null_x). Where p_x is the probability of an amino acid residue, at a particular position in the alignment, according to the Profile HMM and null_x is the probability according to the Null model. The Null model is a simple one state probabilistic model with pre-calculated set of emission probabilities for each of the 20 amino acids derived from the distribution of amino acids in the SWISSPROT release 24. State transition scores are also calculated as log odds parameters and are propotional to Log_2 (t_x). Where t_x is the probability of transiting to an emitter or non-emitter state.
(141) Using a multiple sequence alignment of experimentally verified sequences containing the architecture of interest XFP_N; XFP; XFP_C, a profile Hidden Markov Model (HMM) was created for representing members of the xylulose-5-phosphate/fructose-6-phosphate phosphoketolases (XPK-XFP). As stated in the user guide, Profile HMMs are statistical models of multiple sequence alignments. They capture position-specific information about how conserved each column of the alignment is, and which amino acid residues are most likely to occur at each position. Thus HMMs have a formal probabilistic basis. Profile HMMs for a large number of protein families are publicly available in the PFAM database (Janelia Farm Research Campus, Ashburn, Va.), see ftp://ftp.sanger.ac.uk/pub/databases/Pfam/releases/Pfam24.0/.
(142) Eight xylulose-5-phosphate/fructose-6-phosphate phosphoketolases sequences with experimentally verified function were identified in the BRENDA database: 1. CBF76492.1 from Aspergillus nidulans FGSC A4 (SEQ ID NO: 355) 2. AAR98787.1 from Bifidobacterium longum (SEQ ID NO: 379) 3. ZP_03646196.1 from Bifidobacterium bifidum NCIMB 41171 (SEQ ID NO: 381) 4. ZP_02962870.1 from Bifidobacterium animalis subsp. lactis HNO19 (SEQ ID NO: 388) 5. ZP_786060.1 from Lactobacillus plantarum WCFS1 (SEQ ID NO: 481) 6. ZP_03940142.1 from Lactobacillus brevis subsp. gravesensis ATCC 27305 (SEQ ID NO: 486) 7. ZP_03073172.1 from Lactobacillus reuteri 100-23 (SEQ ID NO 468) 8. YP_818922.1 from Leuconostoc mesenteroides subsp. mesenteroides ATCC 8293 (SEQ ID NO: 504)
The BRENDA database is a freely available information system containing biochemical and molecular information on all classified enzymes as well as software tools for querying the database and calculating molecular properties. The database covers information on classification and nomenclature, reaction and specificity, functional parameters, occurrence, enzyme structure and stability, mutants and enzyme engineering, preparation and isolation, the application of enzymes, and ligand-related data. (BRENDA, AMENDA and FRENDA the enzyme information system: new content and tools in 2009, Nucleic Acids Res. 2009 Jan; 37 (Database issue): D588-92. Epub 2008 Nov. 4. Chang A. Scheer M, Grote A, Schomburg I, Schomburg D.) The eight sequences were used to build a profile HMM which is provided herein as Table 6.
(143) To further identify the proteins of interest, the 436 sequences were searched with tour profile HMMs: the generated XPK_XFP_HMM profile HMM provided in Table 6 as well as the three published profiles for the three domains XFP_N; XFP; XFP_C (PFAM DATABASE) described in Tables 7, 8, and 9, respectively. 309 protein sequences which lengths were between 650 amino acids and 900 amino acids, and contained the three domains were retained.
(144) All 309 sequences are at least 40% identical to an experimentally verified phosphoketolase, with exception of 12 sequences that are within 35% identity distance. However, all 31.9 sequences have a highly significant match to all 4 profile HMMs. The least significant matches have Evalues of 7.5E-242, 1,1E-124, 2,1E-49, 7.8E-37 to XFP_XPK HMM, XFP_N, XFP, and XFP_C profile HMMs respectively. The 309 sequences are provided in Table 5, however, it is understood that any xylulose-5-phosphate/fructose-6-phosphate phosphoketolase identifiable by the method described may be expressed in host cells as described herein. Where accession information is given as complement (NN_NNNNN.N:X . . . Y), it should be understood to mean the reverse complement of nucleotides X to Y of the sequence with Accession number NN_NNNNNN.N. Where accession information is given as join (NNNNNN.N:X . . . Y, NNNNNN.N:Z . . . Q), it should be understood to mean the sequence resulting from joining nucleotides X to Y of NNNNNN.N to nucleotides Z to Q of NNNNNN.N.
(145) TABLE-US-00005 TABLE 5 SEQ ID NOs of xylulose-5-phosphate/fructose-6-phosphate phosphoketolase target gene coding regions and proteins. GENBANK Nucleic GENBANK Amino Nucleotide Acid Amino Acid Acid GI Sequence SEQ Sequence SEQ Source Number Accession Information ID NO: Accession No. ID NO: Organism 162147402 complement 21 YP_001601863.1 330 Gluconacetobacter (NC_010125.1: diazotrophicus 1624982 . . . 1627414) PAl 5 127512024 complement 22 YP_001093221.1 331 Shewanella (NC_009092.1: loihica 1256548 . . . 1258914) PV-4 119774052 complement 23 YP_926792.1 332 Shewanella (NC_008700.1: amazonensis 1105589 . . . 1107955) SB2B 113971300 NC_008321.1: 24 YP_735093.1 333 Shewanella 3541276 . . . 3543642 sp. MR-4 126173290 complement 25 YP_001049439.1 334 Shewanella (NC_009052.1: baltica 1204351 . . . 1206717) OS155 163750647 complement 26 ZP_02157884.1 335 Shewanella (NZ_ABIC01000018.1: benthica 47875 . . . 50253) KT99 157374325 complement 27 YP_001472925.1 336 Shewanella (NC_009831.1: sediminis 1410491 . . . 1412857) HAW-EB3 170725643 complement 28 YP_001759669.1 337 Shewanella (NC_010506.1: woodyi 1624098 . . . 1626464) ATCC 51908 167623058 complement 29 YP_001673352.1 338 Shewanella (NC_010334.1: halifaxensis 1364332 . . . 1366698) HAW-EB4 91794082 NC_007954.1: 30 YP_563733.1 339 Shewanella 3270590 . . . 3272956 denitrificans OS217 254498997 NZ_ACUL01000224.1: 31 ZP_05111697.1 340 Legionella 8243 . . . 10492 drancourtii LLAP12 239607320 join(EQ999973.1: 32 EEQ84307.1 341 Ajellomyces 7995808 . . . 7995885, dermatitidis EQ999973.1: ER-3 7996059 . . . 7996116, EQ999973.1: 7996178 . . . 7996412, EQ999973.1: 7996486 . . . 7996638, EQ999973.1: 7996791 . . . 7997016, EQ999973.1: 7997082 . . . 7997548, EQ999973.1: 7997603 . . . 7997705, EQ999973.1: 7997779 . . . 7998604, EQ999973.1: 7998677 . . . 7998717) 261200667 XM_002626688.1: 33 XP_002626734.1 342 Ajellomyces 1 . . . 2034 dermatitidis SLH14081 154276328 XM_001538959.1: 34 XP_001539009.1 343 Ajellomyces 1 . . . 2421 capsulatus NAm1 225555843 join(GG663374.1: 35 EEH04133.1 344 Ajellomyces 72330 . . . 72492, capsulatus GG663374.1: G186AR 72695 . . . 72929, GG663374.1: 73005 . . . 73157, GG663374.1: 73216 . . . 74203, GG663374.1: 74274 . . . 75146) 225681974 join(DS544805.1: 36 EEH20258.1 345 Paracoccidioides 2187102 . . . 2187264, brasiliensis DS544805.1: Pb03 2187353 . . . 2187427, DS544805.1: 2187521 . . . 2187755, DS544805.1: 2187839 . . . 2187991, DS544805.1: 2188086 . . . 2188395, DS544805.1: 2188462 . . . 2189082, DS544805.1: 2189147 . . . 2189263, DS544805.1: 2189358 . . . 2189973, DS544805.1: 2190044 . . . 2190084) 226289140 join(DS572750.1: 37 EEH44652.1 346 Paracoccidioides 3105630 . . . 3105792, brasiliensis DS572750.1: Pb18 3106049 . . . 3106283, DS572750.1: 3106367 . . . 3106519, DS572750.1: 3106614 . . . 3106923, DS572750.1: 3107023 . . . 3107610, DS572750.1: 3107675 . . . 3108500, DS572750.1: 3108572 . . . 3108612) 258564014 XM_002582706.1: 38 XP_002582752.1 347 Uncinocarpus 1 . . . 2421 reesii 1704 240108203 join(ACFW01000030.1: 39 EER26377.1 348 Coccidioides 1281918 . . . 1282080, posadasii ACFW01000030.1: C735 delta SOWgp 1282133 . . . 1282207, ACFW01000030.1: 1282266 . . . 1282500, ACFW01000030.1: 1282551 . . . 1282703, ACFW01000030.1: 1282757 . . . 1283066, ACFW01000030.1: 1283132 . . . 1283752, ACFW01000030.1: 1283828 . . . 1284653, ACFW01000030.1: 1284726 . . . 1284763) 238838423 join(DS995701.1: 40 EEQ28085.1 349 Microsporum 2675053 . . . 2675215, canis DS995701.1: CBS 113480 2675278 . . . 2675352, DS995701.1: 2675424 . . . 2675658, DS995701.1: 2675745 . . . 2675897, DS995701.1: 2675972 . . . 2676281, DS995701.1: 2676341 . . . 2676961, DS995701.1: 2677062 . . . 2677836, DS995701.1: 2677864 . . . 2677933, DS995701.1: 2677998 . . . 2678013) 169770631 XM_001819733.1: 41 XP_001819785.1 350 Aspergillus 1 . . . 2457 oryzae RIB40 145232813 XM_001399743.1: 42 XP_001399780.1 351 Aspergillus 1 . . . 2421 niger 119491775 XM_001263381.1: 43 XP_001263382.1 352 Neosartorya 1 . . . 2421 fischeri NRRL 181 121705634 XM_001271079.1: 44 XP_001271080.1 353 Aspergillus 1 . . . 2421 clavatus NRRL 1 115396290 XM_001213784.1: 45 XP_001213784.1 354 Aspergillus 1 . . . 2400 terreus NIH2624 259482219 join(BN001303.1: 46 CBF76492.1 355 Aspergillus 576345 . . . 576507, nidulans BN001303.1: FGSC A4 576696 . . . 576930, BN001303.1: 576981 . . . 577133, BN001303.1: 577185 . . . 577494, BN001303.1: 577544 . . . 578161, BN001303.1: 578210 . . . 579035, BN001303.1: 579091 . . . 579128) 255942289 XM_002561867.1: 47 XP_002561913.1 356 Penicillium 1 . . . 2469 chrysogenum Wisconsin 54-1255 242784458 XM_002480346.1: 48 XP_002480391.1 357 Talaromyces 69 . . . 2489 stipitatus ATCC 10500 212527714 XM_002143978.1: 49 XP_002144014.1 358 Penicillium 139 . . . 2559 marneffei ATCC 18224 70999652 XM_749450.1: 50 XP_754543.1 359 Aspergillus 1 . . . 2145 fumigatus Af293 154314622 XM_001556585.1: 51 XP_001556635.1 360 Botryotinia 1 . . . 2061 fuckeliana B05.10 156053245 XM_001592499.1: 52 XP_001592549.1 361 Sclerotinia 1 . . . 2430 sclerotiorum 1980 46124351 XM_386729.1: 53 XP_386729.1 362 Gibberella 1 . . . 2418 zeae PH-1 256733824 complement 54 EEU47171.1 363 Nectria (join(GG698897.1: haematococca 220636 . . . 220670, mpVI 77-13-4 GG698897.1: 220723 . . . 221650, GG698897.1: 221704 . . . 222510, GG698897.1: 222576 . . . 222732, GG698897.1: 222783 . . . 223020, GG698897.1: 223072 . . . 223146, GG698897.1: 223199 . . . 223361)) 261354209 join(DS985216.1: 55 EEY16637.1 364 Verticillium 747889 . . . 748126, albo-atrum DS985216.1: VaMs. 102 748174 . . . 748564, DS985216.1: 748620 . . . 748929, DS985216.1: 748985 . . . 749555, DS985216.1: 749607 . . . 749833, DS985216.1: 749946 . . . 750572) 85081035 XM_951556.2: 56 XP_956649.1 365 Neurospora 215 . . . 2662 crassa OR74A 145609083 XM_364271.2: 57 XP_364271.2 366 Magnaporthe 1 . . . 2442 grisea 70-15 171679277 XM_001904550.1: 58 XP_001904585.1 367 Podospora 1 . . . 2316 anserine 169859036 XM_001836107.1: 59 XP_001836159.1 368 Coprinopsis 1 . . . 2418 cinerea okayama7#130 19112755 NM_001021872.1: 60 NP_595963.1 369 Schizosaccharomyces 1 . . . 2478 pombe 213405339 XM_002173405.1: 61 XP_002173441.1 370 Schizosaccharomyces 1 . . . 2469 japonicus yFS275 58267408 XM_570860.1: 62 XP_570860.1 371 Cryptococcus 39 . . . 2594 neoformans var. neoformans JEC21 71018661 XM_754468.1: 63 XP_759561.1 372 Ustilago 1 . . . 2682 maydis 521 254413307 NZ_DS989851.1: 64 ZP_05027078.1 373 Microcoleus 81897 . . . 84338 chthonoplastes PCC 7420 256377454 NC_013093.1: 65 YP_003101114.1 374 Actinosynnema 3941285 . . . 3943633 mirum DSM 43827 221195188 NZ_ACFE01000002.1: 66 ZP_03568244.1 375 Atopobium 241103 . . . 243577 rimae ATCC 49626 257785020 complement 67 YP_003180237.1 376 Atopobium (NC_013203.1: parvulum 1365893 . . . 1368364) DSM 20469 227516879 complement 68 ZP_03946928.1 377 Atopobium (NZ_ACGK01000053.1: vaginae 213016 . . . 215490) DSM 15829 210630184 NZ_ABXJ01000012.1: 69 ZP_03296299.1 378 Collinsella 49466 . . . 51985 stercoris DSM 13279 41056825 AY518215.1: 70 AAR98787.1 379 Bifidobacterium 989 . . . 3466 longum 223467373 NZ_ACCG01000015.1: 71 ZP_03618909.1 380 Bifidobacterium 9765 . . . 12350 breve DSM 20213 224282874 NZ_ABQP01000009.1: 72 ZP_03646196.1 381 Bifidobacterium 218668 . . . 221073 bifidum NCIMB 41171 229817819 complement 73 ZP_04448101.1 382 Bifidobacterium (NZ_ABYS02000004.1: angulatum 901411 . . . 903888) DSM 20098 212716076 complement 74 ZP_03324204.1 383 Bifidobacterium (NZ_ABXY01000011.1: catenulatum 578312 . . . 580789) DSM 16992 41056831 AY518218.1: 75 AAR98790.1 384 Bifidobacterium 1430 . . . 3907 sp. CFAR 172 41056829 AY518217.1: 76 AAR98789.1 385 Bifidobacterium 951 . . . 3428 pullorum 227507561 NZ_ACGF01000124.1: 77 ZP_03937610.1 386 Gardnerella 41655 . . . 44132 vaginalis ATCC 14019 261337317 NZ_ABXB03000001.1: 78 ZP_05965201.1 387 Bifidobacterium 154886 . . . 157366 gallicum DSM 20093 183601500 complement 79 ZP_02962870.1 388 Bifidobacterium (NZ_ABOT01000001.1: animalis 194894 . . . 197371) subsp. lactis HN019 41056827 AY518216.1: 80 AAR98788.1 389 Bifidobacterium 988 . . . 3465 pseudolongum subsp. Globosum 227516469 complement 81 ZP_03946518.1 390 Atopobium (NZ_ACGK01000047.1: vaginae 28634 . . . 31102) DSM 15829 76556241 AJ509177.1: 82 YP_001511171.1 391 Frankia 1 . . . 2625 sp. EAN1pec 158318663 NC_009921.1: 83 YP_713678.1 392 Frankia 8441355 . . . 8443790 alni ACN14a 111222884 complement 84 YP_002778395.1 393 Rhodococcus (NC_008278.1: opacus 3758441 . . . 3760909) B4 226360617 complement 85 YP_701466.1 394 Rhodococcus (NC_012522.1: jostii 1273076 . . . 1275478) RHA1 111018494 complement 86 ZP_04383880.1 395 Rhodococcus (NC_008268.1: erythropolis 1575800 . . . 1578352) SK121 229490027 NZ_ACNO01000014.1: 87 YP_947598.1 396 Arthrobacter 107516 . . . 109885 aurescens TC1 119962524 NC_008711.1: 88 CAD48946.1 397 Propionibacterium 2018415 . . . 2020796 freudenreichii subsp. Shermanii 28868876 NC_004578.1: 89 NP_791495.1 398 Pseudomonas 1837381 . . . 1839888 syringae pv. tomato str. DC3000 256425339 NC_013132.1: 90 YP_003125992.1 399 Chitinophaga 8027760 . . . 8030123 pinensis DSM 2588 161075783 EU223897.1: 91 ABX56639.1 400 Verrucomicrobiae 1 . . . 2430 bacterium V4 218246512 complement 92 YP_002371883.1 401 Cyanothece (NC_011726.1: sp. PCC 8801 1758431 . . . 1760839) 172055269 NC_010547.1: 93 YP_001806596.1 402 Cyanothece 390265 . . . 392673 sp. ATCC 51142 126659520 complement 94 ZP_01730652.1 403 Cyanothece (NZ_AAXW01000034.1: sp. CCY0110 5415 . . . 7823) 258380665 complement 95 CAQ48286.1 404 Planktothrix (AM990467.1: rubescens 2704 . . . 5112) NIVA-CYA 98 209527806 NZ_ABYK01000067.1: 96 ZP_03276298.1 405 Arthrospira 8063 . . . 10480 maxima CS-328 196258744 NZ_ABVE01000007.1: 97 ZP_03157277.1 406 Cyanothece 72906 . . . 75314 sp. PCC 7822 218440702 complement 98 YP_002379031.1 407 Cyanothece (NC_011729.1: sp. PCC 7424 4207741 . . . 4210149) 166366228 complement 99 YP_001658501.1 408 Microcystis (NC_010296.1: aeruginosa 3156762 . . . 3159182) NIES-843 119488765 NZ_AAVU01000020.1: 100 ZP_01621774.1 409 Lyngbya 110903 . . . 113317 sp. PCC 8106 17228976 complement 101 NP_485524.1 410 Nostoc (NC_003272.1: sp. PCC 7120 1746056 . . . 1748482) 254422632 NZ_DS989904.1: 102 ZP_05036350.1 411 Synechococcus 4613864 . . . 4616290 sp. PCC 7335 158333641 NC_009925.1: 103 YP_001514813.1 412 Acaryochloris 422232 . . . 424652 marina MBIC11017 254425820 complement 104 ZP_05039537.1 413 Synechococcus (NZ_DS989905.1: sp. PCC 7335 71540 . . . 74017) 170695087 complement 105 ZP_02886235.1 414 Burkholderia (NZ_ABLD01000020.1: graminis 33972 . . . 36368) C4D1M 209515639 complement 106 ZP_03264503.1 415 Burkholderia (NZ_ABYL01000006.1: sp. H160 33232 . . . 35628) 87303015 NZ_AANO01000008.1: 107 ZP_01085819.1 416 Synechococcus 122233 . . . 124656 sp. WH 5701 254431900 complement 108 ZP_05045603.1 417 Cyanobium (NZ_DS990556.1: sp. PCC 7001 2146872 . . . 2149313) 88808134 NZ_AAOK01000002.1: 109 ZP_01123645.1 418 Synechococcus 342081 . . . 344516 sp. WH 7805 148238545 complement 110 YP_001223932.1 419 Synechococcus (NC_009481.1: sp. WH 7803 226771 . . . 229206) 87123187 NZ_AANP01000001.1: 111 ZP_01079038.1 420 Synechococcus 180603 . . . 183032 sp. RS9917 187919971 complement 112 YP_001889002.1 421 Burkholderia (NC_010676.1: phytofirmans 1450148 . . . 1452541) PsJN 91778759 complement 113 YP_553967.1 422 Burkholderia (NC_007952.1: xenovorans 1882080 . . . 1884473) LB400 170690542 NZ_ABLD01000001.1: 114 ZP_02881709.1 423 Burkholderia 565487 . . . 567880 graminis C4D1M 209521856 NZ_ABYL01000194.1: 115 ZP_03270532.1 424 Burkholderia 6778 . . . 9171 sp. H160 186474278 complement 116 YP_001861620.1 425 Burkholderia (NC_010623.1: phymatum 2647064 . . . 2649448) STM815 225873826 complement 117 YP_002755285.1 426 Acidobacterium (NC_012483.1: capsulatum 2598033 . . . 2600420) ATCC 51196 206602403 DS995260.1: 118 EDZ38884.1 427 Leptospirillum 236338 . . . 238704 sp. Group II 5-way CG 251772639 complement 119 EES53204.1 428 Leptospirillum (GG693868.1: ferrodiazotrophum 86578 . . . 88956) 56752022 complement 120 YP_172723.1 429 Synechococcus (NC_006576.1: elongatus 2156604 . . . 2158994) PCC 630 22298729 complement 121 NP_681976.1 430 Thermosynechococcus (NC_004113.1: elongatus 1224195 . . . 1226633) BP-1 53804073 NC_002977.6: 122 YP_114037.1 431 Methylococcus 1693459 . . . 1695894 capsulatus str. Bath 220907266 NC_011884.1: 123 YP_002482577.1 432 Cyanothece 1725657 . . . 1728098 sp. PCC 7425 16332268 NC_000911.1: 124 NP_442996.1 433 Synechocystis 3500713 . . . 3503178 sp. PCC 6803 220907424 complement 125 YP_002482735.1 434 Cyanothece (NC_011884.1: sp. PCC 7425 1898702 . . . 1901167) 241777601 complement 126 ZP_04774866.1 435 Allochromatium (NZ_ACQQ01000020.1: vinosum 30393 . . . 32762) DSM 180 114778289 NZ_AATS01000014.1: 127 ZP_01453148.1 436 Mariprofundus 23435 . . . 25801 ferrooxydans PV-1 251827471 complement 128 ZP_04830548.1 437 Gallionella (NZ_ACSD01000006.1: ferruginea 39617 . . . 41986) ES-2 121712503 XM_001273862.1: 129 XP_001273863.1 438 Aspergillus 1 . . . 2436 clavatu NRRL 1 119473535 XM_001258642.1: 130 XP_001258643.1 439 Neosartorya 1 . . . 2439 fischer NRRL 181 169763560 XM_001727628.1: 131 XP_001727680.1 440 Aspergillus 1 . . . 2433 oryzae RIB40 145248115 XM_001396269.1: 132 XP_001396306.1 441 Aspergillus 1 . . . 2448 niger 115400974 XM_001216075.1: 133 XP_001216075.1 442 Aspergillus 1 . . . 2457 terreus NIH2624 255952755 XM_002567084.1: 134 XP_002567130.1 443 Penicillium 1 . . . 2433 chrysogenum Wisconsin 54-1255 212527388 XM_002143815.1: 135 XP_002143851.1 444 Penicillium 98 . . . 2551 marneffei ATCC 18224 242783584 XM_002480171.1: 136 XP_002480216.1 445 Talaromyces 1 . . . 2448 stipitatus ATCC 10500 154321267 XM_001559899.1: 137 XP_001559949.1 446 Botryotinia 1 . . . 2466 fuckeliana B05.10 156054348 XM_001593050.1: 138 XP_001593100.1 447 Sclerotinia 1 . . . 2499 sclerotiorum 1980 189191706 XM_001932157.1: 139 XP_001932192.1 448 Pyrenophora 1 . . . 2469 tritici-repentis Pt-1C-BFP 169600613 XM_001793677.1: 140 XP_001793729.1 449 Phaeosphaeria 1 . . . 2466 nodorum SN15 58260732 XM_567776.1: 141 XP_567776.1 450 Cryptococcus 41 . . . 2545 neoformans var. neoformans JEC21 46123901 XM_386504.1: 142 XP_386504.1 451 Gibberella 1 . . . 2460 zeae PH-1 256732917 complement 143 EEU46265.1 452 Nectria (join(GG698898.1: haematococca 321524 . . . 322233, mpV 77-13-4 GG698898.1: 322285 . . . 322489, GG698898.1: 322540 . . . 324081)) 225729111 FJ790496.1: 144 ACO24516.1 453 Metarhizium 215 . . . 2677 anisopliae 85094948 XM_954892.2: 145 XP_959985.1 454 Neurospora 155 . . . 2638 crassa OR74A 171679479 XM_001904651.1: 146 XP_001904686.1 455 Podospora 1 . . . 2517 anserine 198283820 NC_011206.1: 147 YP_002220141.1 456 Acidithiobacillus 1682860 . . . 1685307 ferrooxidans ATCC 53993 148243889 NC_009468.1: 148 YP_001220128.1 457 Acidiphilium 90683 . . . 93145 cryptum JF-5 157364435 NC_009828.1: 149 YP_001471202.1 458 Thermotoga 1658895 . . . 1661258 lettingae TMO 217966781 NC_011661.1: 150 YP_002352287.1 459 Dictyoglomus 369050 . . . 371428 turgidum DSM 672 92109503 complement 151 YP_571790.1 460 Nitrobacter (NC_007960.1: hamburgensis 14429 . . . 16810) X14 87310270 complement 152 ZP_01092401.1 461 Blastopirellula (NZ_AANZ01000017.1: marina 80191 . . . 82560) DSM 3645 152995974 NC_009654.1: 153 YP_001340809.1 462 Marinomonas 2214232 . . . 2216625 sp. MWYL1 32473390 NC_005027.1: 154 NP_866384.1 463 Rhodopirellula 2520925 . . . 2523306 baltica SH 1 254495580 complement 155 ZP_05108502.1 464 Legionella (NZ_ACUL01000002.1: drancourtii 21176 . . . 23557) LLAP12 254380451 NZ_DS570384.1: 156 ZP_04995817.1 465 Streptomyces 88623 . . . 90992 sp. Mg1 227974767 NZ_ACGW01000133.1: 157 ZP_04023055.1 466 Lactobacillus 1172 . . . 3235 reuter SD2112 227530011 NZ_ACGV01000134.1: 158 ZP_03960060.1 467 Lactobacillus 2320 . . . 4794 vaginalis ATCC 49540 194467185 complement 159 ZP_03073172.1 468 Lactobacillus (NZ_AAPZ02000001.1: reuter 905298 . . . 907709) 100-23 256847586 NZ_GG698803.1: 160 ZP_05553031.1 469 Lactobacillus 21616 . . . 24015 coleohominis 101-4 CHN 260662452 complement 161 ZP_05863347.1 470 Lactobacillus (NZ_GG704700.1: fermentum 145244 . . . 147643) 28-3-CHN 227903484 NZ_ACHN01000046.1: 162 ZP_04021289.1 471 Lactobacillus 59035 . . . 61452 acidophilus ATCC 4796 227877116 NZ_ACKR01000020.1: 163 ZP_03995194.1 472 Lactobacillus 11753 . . . 14191 crispatus JV-V01 227893117 NZ_ACGU01000035.1: 164 ZP_04010922.1 473 Lactobacillus 36787 . . . 39186 ultunensis DSM 16047 256844475 NZ_GG698762.1: 165 ZP_05549961.1 474 Lactobacillus 280846 . . . 283242 crispatus 125-2-CHN 227521312 complement 166 ZP_03951361.1 475 Lactobacillus (NZ_ACGO01000008.1: gasseri 37191 . . . 39647) JV-V03 259501613 complement 167 ZP_05744515.1 476 Lactobacillus (NZ_ACLN01000019.1: iners 10173 . . . 12569) DSM 13335 104773655 NC_008054.1: 168 YP_618635.1 477 Lactobacillus 449229 . . . 451631 delbrueckii subsp. bulgaricus ATCC 11842 227525868 NZ_ACGQ01000037.1: 169 ZP_03955917.1 478 Lactobacillus 36941 . . . 39310 jensenii JV-V16 227512366 NZ_ACGH01000107.1: 170 ZP_03942415.1 479 Lactobacillus 31655 . . . 34045 buchneri ATCC 11577 118587374 complement 171 ZP_01544800.1 480 Oenococcus (NZ_AAUV01000059.1: oeni 59008 . . . 61416) ATCC BAA-1163 28379168 complement 172 NP_786060.1 481 Lactobacillus (NC_004567.1: plantarum 2362936 . . . 2365302) WCFS1 21363093 AJ309011.1: 173 Q937F6 482 XPKA_LACPE 181 . . . 2547 81427904 NC_007576.1: 174 YP_394903.1 483 Lactobacillus 286496 . . . 288859 sakei subsp. sakei 23K 116492156 NC_008525.1: 175 YP_803891.1 484 Pediococcus 398927 . . . 401290 pentosaceus ATCC 25745 259648565 AP011548.1: 176 BAI40727.1 485 Lactobacillus 211570 . . . 213957 rhamnosus GG 227510093 complement 177 ZP_03940142.1 486 Lactobacillus (NZ_ACGG01000115.1: brevis 64541 . . . 66952) subsp. gravesensis ATCC 27305 227891468 complement 178 ZP_04009273.1 487 Lactobacillus (NZ_ACGT01000007.1: salivarius 44265 . . . 46625) ATCC 11741 227528594 NZ_ACGS01000122.1: 179 ZP_03958643.1 488 Lactobacillus 352 . . . 2721 ruminis ATCC 25644 229542373 complement 180 ZP_04431433.1 489 Bacillus (NZ_AAWV02000001.1: coagulans 1384102 . . . 1386486) 36D1 238021480 complement 181 ZP_04601906.1 490 Kingella (NZ_ACJW02000002.1: oralis 913355 . . . 915730) ATCC 51147 259046526 NZ_ACKZ01000008.1: 182 ZP_05736927.1 491 Granulicatella 36586 . . . 38955 adiacens ATCC 49175 157150221 NC_009785.1: 183 YP_001449631.1 492 Streptococcus 333239 . . . 335623 gordonii str. Challis substr. CH1 25011879 complement 184 NP_736274.1 493 Streptococcus (NC_004368.1: agalactiae 1900754 . . . 1903132) NEM316 229555065 complement 185 ZP_04442854.1 494 Listeria (NZ_ACCR01000006.1: grayi 74043 . . . 76418) DSM 20601 257866707 NZ_GG670386.1: 186 ZP_05646360.1 495 Enterococcus 478278 . . . 480644 casseliflavus EC30 257870669 NZ_GG670289.1: 187 ZP_05650322.1 496 Enterococcus 233512 . . . 235875 gallinarum EG2 257895654 NZ_GG670306.1: 188 ZP_05675307.1 497 Enterococcus 612981 . . . 615353 faecium Com12 238810139 AP009608.1: 189 BAH69929.1 498 Mycoplasma 744956 . . . 747334 fermentans PG18 193216764 NC_011025.1: 190 YP_002000006.1 499 Mycoplasma 384420 . . . 386801 arthritidis 158L3-1 148377390 NC_009497.1: 191 YP_001256266.1 500 Mycoplasma 136795 . . . 139182 agalactiae PG2 191639669 NC_010999.1: 192 YP_001988835.1 501 Lactobacillus 2885324 . . . 2887711 casei BL23 28379861 NC_004567.1: 193 NP_786753.1 502 Lactobacillus 3169067 . . . 3171478 plantarum WCFS1 227892171 complement 194 ZP_04009976.1 503 Lactobacillus (NZ_ACGT01000037.1: salivarius 21330 . . . 23759) ATCC 11741 116618551 NC_008531.1: 195 YP_818922.1 504 Leuconostoc 1449343 . . . 1451709 mesenteroides subsp. mesenteroides ATCC 8293 116333142 NC_008497.1: 196 YP_794669.1 505 Lactobacillus 507704 . . . 510163 brevis ATCC 367 241895257 complement 197 ZP_04782553.1 506 Weissella (NZ_ACKU01000007.1: paramesenteroides 101374 . . . 103833) ATCC 33313 170016535 NC_010471.1: 198 YP_001727454.1 507 Leuconostoc 181964 . . . 184417 citreum KM20 116619034 complement 199 YP_819405.1 508 Leuconostoc (NC_008531.1: mesenteroides 1934181 . . . 1936622) subsp. mesenteroides ATCC 8293 161702316 EU255918.1: 200 ABX75772.1 509 Lactococcus 18411 . . . 20879 lactis subsp. Lactis 116491770 complement 201 YP_811314.1 510 Oenococcus (NC_008528.1: oeni 1731509 . . . 1733962) PSU-1 182419955 complement 202 ZP_02951191.1 511 Clostridium (NZ_ABDT01000107.2: butyricum 13616 . . . 15991) 5521 255523324 complement 203 ZP_05390294.1 512 Clostridium (NZ_ACVI01000003.1: carboxidivorans 55354 . . . 57747) P7 15894622 NC_003030.1: 204 NP_347971.1 513 Clostridium 1482782 . . . 1485172 acetobutylicum ATCC 824 226324778 complement 205 ZP_03800296.1 514 Coprococcus (NZ_ABVR01000041.1: comes 500857 . . . 503232) ATCC 27758 253580358 NZ_GG696051.1: 206 ZP_04857624.1 515 Ruminococcus 158015 . . . 160390 sp. 5_1_39B_FAA 257413435 NZ_ABYJ02000055.1: 207 ZP_04743029.2 516 Roseburia 10320 . . . 12779 intestinalis L1-82 154500233 complement 208 ZP_02038271.1 517 Bacteroides (NZ_AAXG02000041.1: capillosus 34174 . . . 36609) ATCC 29799 219119570 XM_002180506.1: 209 XP_002180542.1 518 Phaeodactylum 1 . . . 2508 tricornutum CCAP 1055/1 91975971 NC_007958.1: 210 YP_568630.1 519 Rhodopseudomonas 1660408 . . . 1662762 palustris BisB5 86750966 complement 211 YP_487462.1 520 Rhodopseudomonas (NC_007778.1: palustris 4411322 . . . 4413676) HaA2 39934743 NC_005296.1: 212 NP_947019.1 521 Rhodopseudomonas 1858439 . . . 1860790 palustris CGA009 90425290 complement 213 YP_533660.1 522 Rhodopseudomonas (NC_007925.1: palustris 4235875 . . . 4238229) BisB18 121583071 NC_008758.1: 214 YP_973512.1 523 Polaromonas 65532 . . . 67904 naphthalenivorans CJ2 115376972 complement 215 ZP_01464191.1 524 Stigmatella (NZ_AAMD01000095.1: aurantiaca 4173 . . . 6533) DW4/3-1 148547676 complement 216 YP_001267778.1 525 Pseudomonas (NC_009512.1: putid 2807645 . . . 2810020) F1 116668711 NC_008541.1: 217 YP_829644.1 526 Arthrobacter 145493 . . . 147928 sp. FB24 220911083 NC_011886.1: 218 YP_002486392.1 527 Arthrobacter 321712 . . . 324174 chlorophenolicus A6 260517200 complement 219 ZP_05816651.1 528 Sanguibacter (NZ_ABUN01000002.1: keddieii 92936 . . . 95461) DSM 10542 229821527 NC_012669.1: 220 YP_002883053.1 529 Beutenbergia 3398743 . . . 3401217 cavernae DSM 12333 256832813 NC_013174.1: 221 YP_003161540.1 530 Jonesia 1712156 . . . 1714588 denitrifican DSM 20603 227428425 complement 222 ZP_03911482.1 531 Xylanimonas (NZ_ABVC01000008.1: cellulosilytica 152502 . . . 154988) DSM 15894 165929357 AM182260.1: 223 CAJ57850.1 532 Cellulomonas 1 . . . 2481 flavigena 145223927 NC_009338.1: 224 YP_001134605.1 533 Mycobacterium 3525804 . . . 3528275 gilvum PYR-GCK 120404048 NC_008726.1: 225 YP_953877.1 534 Mycobacterium 3236585 . . . 3239083 vanbaalenii PYR-1 257069356 NC_013172.1: 226 YP_003155611.1 535 Brachybacterium 2493744 . . . 2496215 faecium DSM 4810 256824167 NC_013169.1: 227 YP_003148127.1 536 Kytococcus 273585 . . . 276047 sedentarius DSM 20547 148271607 NC_009480.1: 228 YP_001221168.1 537 Clavibacter 506602 . . . 509040 michiganensis subsp. michiganensis NCPPB 382 145594129 complement 229 YP_001158426.1 538 Salinispora (NC_009380.1: tropica 1798516 . . . 1800918) CNB-440 159037167 complement 230 YP_001536420.1 539 Salinispora (NC_009953.1: arenicola 1767167 . . . 1769569) CNS-205 238063593 complement 231 ZP_04608302.1 540 Micromonospora (NZ_GG657738.1: sp ATCC 39149 5405062 . . . 5407251) 118469963 NC_008596.1: 232 YP_887914.1 541 Mycobacterium 3674267 . . . 3676639 smegmatis str. MC2 155 108799759 NC_008146.1: 233 YP_639956.1 542 Mycobacterium 2939527 . . . 2941947 sp. MCS 240170498 complement 234 ZP_04749157.1 543 Mycobacterium (NZ_ACBV01000039.1: kansasii 21 . . . 2423) ATCC 12478 183982748 complement 235 YP_001851039.1 544 Mycobacterium (NC_010612.1: marinum 3341817 . . . 3344219) M 41407671 complement 236 NP_960507.1 545 Mycobacterium (NC_002944.2: avium 1726717 . . . 1729131) subsp. paratuberculosis K-10 254819329 NZ_ABIN01000047.1: 237 ZP_05224330.1 546 Mycobacterium 36474 . . . 38837 intracellulare ATCC 13950 169629591 complement 238 YP_001703240.1 547 Mycobacterium (NC_010397.1: abscessus 2559451 . . . 2561871) 84496279 complement 239 ZP_00995133.1 548 Janibacter (NZ_AAMN01000002.1: sp. HTCC2649 433314 . . . 435707) 72163369 NC_007333.1: 240 YP_291026.1 549 Thermobifida 3478272 . . . 3480650 fusca YX 227984600 complement 241 ZP_04031845.1 550 Thermomonospora (NZ_ABUZ01000013.1: curvata 70531 . . . 72909) DSM 43183 229855558 complement 242 ZP_04475514.1 551 Streptosporangium (NZ_ABUU01000106.1: roseum 1385 . . . 3700) DSM 43021 229209207 NZ_ABUI01000028.1: 243 ZP_04335641.1 552 Nocardiopsis 55841 . . . 58189 dassonvillei subsp. dassonvillei DSM 43111 229862587 NZ_ABUS01000001.1: 244 ZP_04482201.1 553 Stackebrandtia 1911934 . . . 1914330 nassauensis DSM 44728 256376052 NC_013093.1: 245 YP_003099712.1 554 Actinosynnema 2125566 . . . 2127935 mirum DSM 43827 32141117 complement 246 NP_733508.1 555 Streptomyces (NC_003888.3: coelicolor 656319 . . . 658772) A3(2) 117164830 complement 247 CAJ88379.1 556 Streptomyces (AM238664.1: ambofaciens 846551 . . . 849055) ATCC 23877 256811868 complement 248 ZP_05536883.1 557 Streptomyces (NZ_ACFA01000015.1: griseoflavus 3377 . . . 5761) Tu400 254405496 complement 249 ZP_05020421.1 558 Streptomyces (NZ_DS570938.1: sviceus 43288 . . . 45726) ATCC 29083 260644540 complement 250 CBG67625.1 559 Streptomyces (FN554889.1: scabiei 480316 . . . 482694) 87.22 29827814 complement 251 NP_822448.1 560 Streptomyces (NC_003155.4: avermitilis 1579336 . . . 1581717) MA-4680 239932594 NZ_ABYA01000503.1: 252 ZP_04689547.1 561 Streptomyces 5217 . . . 7595 ghanaensis ATCC 14672 256800397 complement 253 ZP_05530021.1 562 Streptomyces (NZ_ACEZ01000048.1: viridochromogenes 24916 . . . 27303) DSM 40736 256774038 complement 254 ZP_05512501.1 563 Streptomyces (NZ_ACEX01000074.1: hygroscopicus 46221 . . . 48614) ATCC 53653 260452518 NZ_ACZH01000001.1: 255 ZP_05800927.1 564 Streptomyces 321972 . . . 324359 flavogriseus ATCC 33331 182440556 NC_010572.1: 256 YP_001828275.1 565 Streptomyces 8084439 . . . 8086826 griseus subsp. griseus NBRC 13350 239982969 NZ_ABYC01000425.1: 257 ZP_04705493.1 566 Streptomyces 13265 . . . 15646 albus J1074 254381599 NZ_DS570386.1: 258 ZP_04996963.1 567 Streptomyces 118817 . . . 121204 sp. Mg1 256674998 NZ_ACEU01000020.1: 259 ZP_05485309.1 568 Streptomyces 1507 . . . 3900 sp. SPB78 227377421 NZ_ABUC01000002.1: 260 ZP_03860882.1 569 Kribbella 229225 . . . 231603 flavida DSM 17836 54023297 complement 261 YP_117539.1 570 Nocardia (NC_006361.1: farcinica 1487629 . . . 1490097) IFM 10152 158313048 NC_009921.1: 262 YP_001505556.1 571 Frankia 1426213 . . . 1428621 sp. EAN1pec 86742227 complement 263 YP_482627.1 572 Frankia (NC_007777.1: sp. CcI3 4238578 . . . 4240986) 256395329 NC_013131.1: 264 YP_003116893.1 573 Catenulispora 7133131 . . . 7135533 acidiphila DSM 44928 117927729 NC_008578.1: 265 YP_872280.1 574 Acidothermus 555555 . . . 557948 cellulolyticus 11B 119717842 complement 266 YP_924807.1 575 Nocardioides (NC_008699.1: sp. JS614 3839565 . . . 3841961) 134098496 NC_009142.1: 1267 YP_001104157.1 576 Saccharopolyspora 2098116 . . . 2100512 erythraea NRRL 2338 209550756 NC_011369.1: 268 YP_002282673.1 577 Rhizobium 3264963 . . . 3267347 leguminosarum bv. trifolii WSM2304 241206160 NC_012850.1: 269 YPJ002977256.1 578 Rhizobium 3503904 . . . 3506288 leguminosarum bv. trifolii WSM1325 190893254 NC_010994.1: 270 YP_001979796.1 579 Rhizobium 3714233 . . . 3716620 etli CIAT 652 86359034 NC_007761.1: 271 YP_470926.1 580 Rhizobium 3623921 . . . 3626308 etli CF42 222081270 complement 272 YP_002540633.1 581 Agrobacterium (NC_011983.1: radiobacter 490969 . . . 493383) K84 254720555 NZ_ACBQ01000064.1: 273 ZP_05182366.1 582 Brucella 129340 . . . 131718 sp. 83/13 239835057 complement 274 ZP_04683384.1 583 Ochrobactrum (NZ_ACQA01000003.1: intermedium 10528 . . . 13017) LMG 3301 153012043 NC_009671.1: 275 YP_001373254.1 584 Ochrobactrum 16319 . . . 18706 anthropi ATCC 49188 146339061 complement 276 YP_001204109.1 585 Bradyrhizobium (NC_009445.1: sp. ORS278 2141749 . . . 2144226) 148253833 complement 277 YP_001238418.1 586 Bradyrhizobium (NC_009485.1: sp. BTAi1 2424642 . . . 2427059) 27377629 complement 278 NP_769158.1 587 Bradyrhizobium (NC_004463.1: japonicum 2749734 . . . 2752139) USDA 110 92117435 complement 279 YP_577164.1 588 Nitrobacter (NC_007964.1: hamburgensis 2109162 . . . 2111570) X14 240137143 NC_012808.1: 280 YP_002961612.1 589 Methylobacterium 407982 . . . 410417 extorquens AM1 110634584 complement 281 YP_674792.1 590 Mesorhizobium (NC_008254.1: sp. BNC1 2388345 . . . 2390747) 260467447 NZ_ACZA01000051.1: 282 ZP_05813617.1 591 Mesorhizobium 15952 . . . 18360 opportunistum WSM2075 75676138 NC_007406.1: 283 YP_318559.1 592 Nitrobacter 2135469 . . . 2137856 winogradskyi Nb-255 170749020 complement 284 YP_001755280.1 593 Methylobacterium (NC_010505.1: radiotolerans 2769888 . . . 2772470) JCM 2831 170746859 complement 285 YP_001753119.1 594 Methylobacterium (NC_010505.1: radiotolerans 465997 . . . 468552) JCM 2831 254558916 NC_012988.1: 286 YP_003066011.1 595 Methylobacterium 271224 . . . 273809 extorquens DM4 240140298 NC_012808.1: 287 YP_002964777.1 596 Methylobacterium 3931130 . . . 3933676 extorquens AM1 220925990 NC_011894.1: 288 YP_002501292.1 597 Methylobacterium 6291823 . . . 6294321 nodulans ORS 2060 220919962 complement 289 YP_002495265.1 598 Methylobacterium (NC_011892.1: nodulans 451339 . . . 453840) ORS 2060 170741732 NC_010511.1: 290 YP_001770387.1 599 Methylobacterium 3988668 . . . 3991166 sp. 4-46 239815802 complement 291 YP_002944712.1 600 Variovorax (NC_012791.1: paradoxus 2971257 . . . 2973608) S110 89069402 NZ_AAOT01000017.1: 292 ZP_01156757.1 601 Oceanicola 31124 . . . 33460 granulosus HTCC2516 119509641 complement 293 ZP_01628787.1 602 Nodularia (NZ_AAVW01000007.1: spumigena 15878 . . . 18259) CCY9414 186682350 NC_010628.1: 294 YP_001865546.1 603 Nostoc 2389837 . . . 2392218 punctiforme PCC 73102 75906719 complement 295 YP_321015.1 604 Anabaena (NC_007413.1: variabilis 617971 . . . 620352) ATCC 29413 225522346 NZ_ACIR01000182.1: 296 ZP_03769140.1 605 Nostoc 624 . . . 2756 azollae 070 37520566 NC_005125.1: 297 NP_923943.1 606 Gloeobacter 1065716 . . . 1068097 violaceus PCC 7421 86608623 NC_007776.1: 298 YP_477385.1 607 Synechococcus 1182311 . . . 1184686 sp. JA-2- 3Ba(2-13) 150398192 complement 299 YP_001328659.1 608 Sinorhizobium (NC_009636.1: medicae WSM419 3144485 . . . 3146857) 116249832 complement 300 YP_765670.1 609 Rhizobium (NC_008380.1: leguminosarum 82152 . . . 84617) bv. viciae 3841 195970218 complement 301 NP_384212.2 610 Sinorhizobium (NC_003047.1: meliloti 123688 . . . 126141) 1021 171912985 NZ_ABIZ01000001.1: 302 ZP_02928455.1 611 Verrucomicrobium 4370841 . . . 4373354 spinosum DSM 4136 163849496 complement 303 YP_001637539.1 612 Methylobacterium (NC_010172.1: extorquens 46285 . . . 48720) PA1 85714839 NZ_AAMY01000005.1: 304 ZP_01045825.1 613 Nitrobacter 72844 . . . 75210 sp. Nb-311A 168704325 complement 305 ZP_02736602.1 614 Gemmata (NZ_ABGO01000323.1: obscuriglobus 57 . . . 2462) UQM 2246 256829143 complement 306 YP_003157871.1 615 Desulfomicrobium (NC_013173.1: baculatum 1530488 . . . 1532881) DSM 4028 223939426 complement 307 ZP_03631304.1 616 bacterium (NZ_ABOX02000044.1: Ellin514 41392 . . . 43863) 237747078 complement 308 ZP_04577558.1 617 Oxalobacter (NZ_GG658151.1: formigenes 2005797 . . . 2008190) HOxBLS 237749232 complement 309 ZP_04579712.1 618 Oxalobacter (NZ_GG658170.1: formigenes 2042015 . . . 2044411) OXCC13 116624013 NC_008536.1: 310 YP_826169.1 619 Solibacter 6218168 . . . 6220537 usitatus Ellin6076 194336959 complement 311 YP_002018753.1 620 Pelodictyon (NC_011060.1: phaeoclathratiforme 2004498 . . . 2006885) BU-1 194334425 complement 312 YP_002016285.1 621 Prosthecochloris (NC_011059.1: aestuarii 1762093 . . . 1764489) DSM 271 189346840 complement 313 YP_001943369.1 622 Chlorobium (NC_010803.1: limicola 1427679 . . . 1430054) DSM 245 21674344 complement 314 NP_662409.1 623 Chlorobium (NC_002932.3: tepidum 1423776 . . . 1426289) TLS 110597897 complement 315 ZP_01386179.1 624 Chlorobium (NZ_AASE01000009.1: ferrooxidans 37756 . . . 40179) DSM 13031 78187379 complement 316 YP_375422.1 625 Chlorobium (NC_007512.1: luteolum 1709621 . . . 1712050) DSM 273 71907690 complement 317 YP_285277.1 626 Dechloromonas (NC_007298.1: aromatica 2220090 . . . 2222456) RCB 74316849 NC_007404.1: 318 YP_314589.1 627 Thiobacillus 876540 . . . 878978 denitrificans ATCC 25259 91775246 complement 319 YP_545002.1 628 Methylobacillus (NC_007947.1: flagellatus 935825 . . . 938200) KT 30250069 NC_004757.1: 320 NP_842139.1 629 Nitrosomonas 2318109 . . . 2320481 europaea ATCC 19718 114332052 NC_008344.1: 321 YP_748274.1 630 Nitrosomonas 2209596 . . . 2211971 eutrqpha C91 82702122 NC_007614.1: 322 YP_411688.1 631 Nitrosospira 1152112 . . . 1154535 multiformis ATCC 25196 77166175 NC_007484.1: 323 YP_344700.1 632 Nitrosococcus 3082455 . . . 3084869 oceani ATCC 1970 46445639 complement 324 YP_007004.1 633 Candidatus (NC_005861.1: Protochlamydia 5907 . . . 8303) amoebophila UWE25 16263040 complement 325 NP_435833.1 634 Sinorhizobium (NC_003037.1: meliloti 591065 . . . 593440) 1021 229532493 NZ_ABUV01000006.1: 326 ZP_04421874.1 635 Sulfurospirillum 47234 . . . 49585 deleyianum DSM 6946 13475490 NC_002678.2: 327 NP_107054.1 636 Mesorhizobium 5384229 . . . 5386652 loti MAFF303099 209885940 complement 328 YP_002289797.1 637 Oligotropha (NC_011386.1: carboxidovorans 2786353 . . . 2788746) OM5 182679166 NC_010581.1: 329 YP_001833312.1 638 Beijerinckia 2524033 . . . 2526420 indica subsp. indica ATCC 9039
(146) Numerous examples of polynucleotides, genes and/or polypeptides encoding phosphotransacetylase are known in the art and can be used in relation to the recombinant host cells disclosed herein. In embodiments, the phosphotransacetylase can be EutD from Lactobacillus plantarum. In embodiments, the phosphotransacetylase can be the phosphotransacetylase from Bacillus subtilis. This phosphotransacetylase has a specific activity of 1371 mmol/min/mg and a Km 0.06 mM for acetyl-CoA (Rado and Hoch, Biochim. Biophys. Acta. 321: 114-25; 1973). In addition, the equilibrium constant (Keq) of this reaction was found to be 15414 in favor of the formation of acetyl-CoA according to the following formula:
(147)
(148) In embodiments, host cells comprise a polypeptide having at least about 80%, at least about 85%, at least about 90%, or at least about 99% identity to a polypeptide of Table 10 or an active fragment thereof or a polynucleotide encoding such a polypeptide. In embodiments, the phosplotransacetylase can be a polypeptide having at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 99% identity to SEQ ID NO: 1472 or an active fragment thereof. In other embodiments, a polynucleotide, gene and/or polypeptide encoding phosphotransacetylase can include, but is not limited to, a sequence provided in the following tables 10 or 12.
(149) TABLE-US-00006 TABLE10 SEQIDNOsofphosphotransacetylasetargetgenecodingregionsandproteins. SEQID SEQID NO: NO: Nucleic Amino Description acid acid Aminoacidsequence EutD 1111 1472 MDLFESLAQKITGKDQTIVFPEGTEPRIVGAAARLAADGLVKPIVLGATDKVQAVANDLN phospho- ADLTGVQVLDPATYPAEDKQAMLDALVERRKGKNTPEQAAKMLEDENYFGTMLVYMGKAD trans- GMVSGAIHPTGDTVRPALQIIKTKPGSHRISGAFIMQKGEERYVFADCAINIDPDADTLA acetylase EIATQSAATAKVFDIDPKVAMLSFSTKGSAKGEMVTKVQEATAKAQAAEPELAIDGELQF fromLacto- DAAFVEKVGLQKAPGSKVAGHANVFVFPELQSGNIGYKIAQRFGHFEAVGPVLQGLNKPV bacillus SDLSRGCSEEDVYKVAIITAAQGLA plantarum Phospho- 1061 1422 MADLFSTVQEKVAGKDVKIVFPEGLDERILEAVSKLAGNKVLNPIVIGNENEIQAKAKEL trans- NLTLGGVKIYDPHTYEGMEDLVQAFVERRKGKATEEQARKALLDENYFGTMLVYKGLADG acetylase LVSGAAHSTADTVRPALQIIKTKEGVKKTSGVFIMARGEEQYVFADCAINIAPDSQDLAE from IAIESANTAKMFDIEPRVAMLSFSTKGSAKSDETEKVADAVKIAKEKAPELTLDGEFQFD Bacillus AAFVPSVAEKKAPDSEIKGDANVFVFPSLEAGNIGYKIAQRLGNFEAVGPILQGLNMPVN subtilis DLSRGCNAEDVYNLALITAAQAL
(150) Additional suitable phosphotransacetylase target gene coding regions and proteins were identified by diversity searching and clustering. A blast search of the non redundant GenBank protein database (NR) was performed with the L. plantarum EutD protein as a query sequence. A blast cut-off (Evalue) of 0.01 was applied. This search resulted in 2124 sequence matches. Redundancy reduction was achieved by clustering proteins with the CD-HIT program with parameters set at 95% sequence identity and 90% length overlap. The longest seed sequence, representative of each cluster, is retained for further analysis. Clustering reduced the number of protein sequences to 1336. Further clean-up of the sequences by removing sequences <280 amino acids and sequences >795 amino acids resulted in 1231 seqs.
(151) The Brenda database was queried for experimentally verified phosphate acetyltransferases. Thirteen were found in the following organisms: S. enterica, E. coli K12, V. Parvula, C. Kluyveri, C. Acetobutylicum, C. Thermocellum, M thermophila, S. pyogenes, B. subtilis, L. fermentum, L. plantarum, L. sanfranciscensis, B subtilis, L. fermentum, L. plantarum, L. sanfranciscensis, R. palustris, E. coli.
(152) Experimentally verified phosphate acetyltransferases (EC 2.3.1.8) belong to the PTA_PTB pfam family. However, the PTA_PTB domain is present in 13 distinct architectures (http://pfam.janelia.org/family/PF01515, Pfam database version 24). The motivation for investigating the domain architecture is to determine which of the proteins, that were identified by BLAST search, are likely to be phosphate acetyltransferases.
(153) Experimentally verified sequences extracted from the BRENDA database as well as sequences retained after the CD-HIT clustering and clean-up, were searched against the Pfam database to determine their domain architecture. Pfam is a collection of multiple sequence alignments and profile hidden Markov models (HMMs). Each Pfam HMM represents a protein family or domain. By searching a protein sequence against the Pfam library of HMMs, it is possible to determine which domains it carries i.e. its domain architecture. (The Pfam protein families database: R. D. Finn, J. Tate, J. Mistry, P. C. Coggill, J. S. Sammut, H. R. Hotz, G. Ceric, K. Forslund, S. R. Eddy, E. L. Sonnhammer and A. Bateman Nucleic Acids Research (2008) Database Issue 36:D281-D288)
(154) Twelve of the experimentally verified proteins only contained the PTA_PTB domain. Two sequences, from R. palustris and E. coli, contained two domains PTA_PTB and DRTGG, a domain of unknown function. Therefore, from the CD-HIT clustering results, proteins that contained either the PTA_PTB domain only (Group 1: 549 sequences) or a combination of PTA_PTB+DRTGG domains (Group 2: 201 sequences) were chosen.
(155) Furthermore, the PTA_PTB domain, as the name indicates, is actually not specific to phosphate acetyltransferases. The domain is also a signature for phosphate butyryltransferases (EC 2.3.1.19) methods to distinguish between the two subfamilies: acetyltransferases and butyryltransferases were employed and are as follows:
(156) To further characterize the relationship among the sequences, multiple sequence alignment MSA), phylogenetic analysis. profile HMMs and GroupSim analysis were performed, For this set of analyses, the phosphate acetyltransferases are split in two groups. Group 1 contains phosphate acetyltransferases with the PTA_PTB domain only, while Group 2 contains phosphate acetyltransferases with PTA_PTB+DRTGG. The motivation here is to generate groups with similar lengths.
(157) Clustal X, version 2.0 was used for sequence alignments with default parameters. (Thompson JD, et al. The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res. (1997) 25:4876-4882.)
(158) Alignment results were utilized to compute % sequence identities to a reference sequence. If the sequence from L. plantarum is taken as a reference, % IDs range from as low as 10.5% to 75.6% for the closest sequence. Alignment results also provided the basis for re-constructing phylogenetic trees. The Neighbor Joining method, available in the Jalview package version 2.3, was used to produce the trees, and computed trees were visualized in MEGA 4 (Tamura, Dudley, Nei, and Kumar 2007). The Neighbor Joining method is a method for re-constructing phylogenetic trees, and computing the lengths of the branches of this tree. In each stage, the two nearest nodes of the tree (the term nearest nodes will be defined in the following paragraphs) are chosen and defined as neighbors in our tree. This is done recursively until all of the nodes are paired together. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol. 1987 July; 4(4):406-25. Saitou N, Nei M. Jalview Version 2 is a system for interactive editing, analysis and annotation of multiple sequence alignments (Waterhouse, A. M., Procter, J. B., Martin, D. M. A, Clamp, M. and Barton, G. J. (2009) Jalview Version 2a multiple sequence alignment editor and analysis workbench Bioinformatics 25 (9) 1189-1191). The MEGA software provides tools for exploring, discovering, and analyzing DNA and protein sequences from an evolutionary perspective. MEGA4 enables the integration of sequence acquisition with evolutionary analysis. It contains an array of input data and multiple results explorers for visual representation; the handling and editing of sequence data, sequence alignments, inferred phylogenetic trees; and estimated evolutionary distances. The results explorers allow users to browse, edit, summarize, export, and generate publication-quality captions for their results. MEGA 4 also includes distance matrix and phylogeny explorers as well as advanced graphical modules for the visual representation of input data and output results (Tamura K, Dudley J, Nei M & Kumar S (2007) MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Molecular Biology and Evolution 24:1596-1599).
(159) Taken together, % IDs and the generated tree (
(160) Based on experimentally verified sequences contained within each of the Subfamilies, Subfamily 1 and Subfamily 2 were determined to represent phosphate butyryltransferases (PTB) and phosphate acetylytransferases (PTA) respectively.
(161) Discrimination between Subfamily 1 members and Subfamily 2 members was also performed by GroupSim analysis (Capra and Singh (2008) Bioinformatics 24: 1473-1480). The GroupSim method identifies amino acid residues that determine a protein's functional specificity. In a multiple sequence alignment (MSA) of a protein family whose sequences are, divided into multiple Subfamilies, amino acid residues that distinguish between the functional Subfamilies of sequences can be identified. The method takes a multiple sequence alignment (MSA) and known specificity groupings as input, and assigns a score to each amino acid position in the MSA. Higher scores indicate a greater likelihood that an amino acid position is a specificity determining position (SDP).
(162) GroupSim analysis performed on the MSA of 537 sequences (divided into Subfamily 1 and Subfamily 2 by the phylogenetic analysis, above) identified highly discriminating positions. Listed in Table 11 are positions (Pos) having scores greater than to 0.7, where, a perfect score of 1.0 would indicate that all proteins within the Subfamily have the listed amino acid in the specified position and between Subfamilies the amino acid would always be different. The Pattern columns give the amino acid(s) in single letter code. Numbers between parentheses indicate frequency of occurrence of each amino acid at the particular position. The amino acid position number in column 1 is for the PTA protein sequence from Lactobacillus plantarum, the representative protein of group 2 with a GI#28377658 (SEQ ID NO: 1472).
(163) TABLE-US-00007 TABLE 11 Highly discriminating amino acid positions for Subfamily 1 (PTB) and Subfamily 2 (PTA) from GroupSim analysis. Pos Score Pattern PTB Pattern PTA 212 0.980314 Group 1: E(173), D(2), Group 2: S(360), N(1) L(1) 305 0.87236 Group 1: L(152), V(11), Group 2: D(360), Q(1) M(5), I(5), F(3) 242 0.831201 Group 1: A(142), D(15), Group 2: Q(361) S(13), G(4), T(2) 208 0.776954 Group 1: L(130), I(35), Group 2: S(355), A(6) V(11) 125 0.705868 Group 1: K(175), R(1) Group 2: S(215), A(85), G(41), C(14), N(4), T(2)
(164) An alternative structure/function characterization of the PTA and PTB subfamilies of enzymes was performed using the HMMER software package (the theory behind profile HMMs is described in R. Durbin, S. Eddy, A. Krogh, and G. Mitchison, Biological sequence analysis: probabilistic models of proteins and nucleic acids, Cambridge University Press, 1998; Krogh et al., 1994; J. Mol. Biol. 235:1501-1531), following the user guide which is available from HMMER (Janelia Farm Research Campus, Ashburn, Va.).
(165) Using a multiple sequence alignment of the experimentally verified sequences (containing the PTA_PTB domain only) in Subfamily 2, a profile Hidden Markov Model (HMM) was created for representing Subfamily 2 members. The sequences were: 1. BAB19267.1 from Lactobacillus sanfranciscensis (SEQ ID NO: 1475) 2. NP_784550.1 from Lactobacillus plantarum WCFS1 (SEQ ID NO: 1472) 3. ZP_03944466.1 from Lactobacillus fermentum ATCC 14931 (SEQ ID NO: 1453) 4. NP_391646.1 from Bacillus subtilis subsp. subtilis str. 168 (SEQ ID NO: 1422) 5. AAA72041.1 from Methanosarcina thermophila (SEQ ID NO: 1277) 6. ZP_03152606.1 from Clostridium thermocellum DSM 4150 (SEQ ID NO: 1275) 7. NP_348368.1 from Clostridium acetobutylicum ATCC 824 (SEQ ID NO: 1206) 8. YP 001394780.1 from Clostridium kluyveri DSM 555 (SEQ ID NO: 1200) 9. ZP_03855267.1 from Veillonella parvula DSM 2008 (SEQ ID NO: 1159) 10. YP_149725.1 from Salmonella enterica subsp. enterica serovar Paratyphi A str. ATCC 9150 (SEQ ID NO: 1129)
(166) The Profile HMM was built as follows: The 10 seed sequences (sequences representing experimentally verified function) that are in Subfamily 2 were aligned using Clustal X (interface to Clustal W) with default parameters. The hmmbuild program was run on each set of the aligned sequences using default parameters. hmmbuild reads the multiple sequence alignment file, builds a new Profile HMM, and saves the Profile HMM to file. Using this program an un-calibrated profile was generated from the multiple alignment for each set of subunit sequences described above.
(167) The Profile HMM was read using hrnmcalibrate which scores a large number of synthesized random sequences with the Profile (the default number of synthetic sequences used is 5,000), fits an extreme value distribution (EVD) to the histogram of those scores, and re-saves the HMM file now including the EVD parameters. These EVD parameters ( and ) are used to calculate the E-values of bit scores when the profile is searched against a protein sequence database. hmmcalibrate writes two parameters into the HMM file on a line labeled EVD: these parameters are the (location) and (scale) parameters of an extreme value distribution (EVD) that best fits a histogram of scores calculated on randomly generated sequences of about the same length and residue composition as SWISS-PROT. This calibration was done once for the Profile HMM.
(168) The calibrated pofile HMM for the Subfamily 2 set is provided as Table 14. The Profile HMM table gives the probability of each amino acid occurring at each position in the amino acid sequence. The amino acids are represented by the one letter code. The first line for each position reports the match emission scores: probability for each amino acid to be in that state (highest score is highlighted). The second line reports the insert emission scores, and the third line reports on state transition scores: M.fwdarw.M, M.fwdarw.I, M.fwdarw.D; I.fwdarw.M, I.fwdarw.I; D.fwdarw.M, D.fwdarw.D; B.fwdarw.M; M.fwdarw.E. Table 14 shows that in the Subfamily 2 profile HMM, methionine has a 3792 ans 4481 probability of being in the first two positions.
(169) The Subfamily 2 profile HMM was evaluated using, hmmsearch, with the Z parameter set to one billion, for the ability to discriminate Subfamily 1 members from those of Subfamily 2. The hmmsearch program takes the hmm file for the Subfamily 2 profile HMM and all the sequences from both Subfamilies and assigns an E-value score to each sequence. This E-value score is a measure of fit to the Profile HMM, with a lower score being a better fit. The Profile HMM distinguished Subfamily 2 members from Subfamily 1 members and there was a large margin of E-value difference between the worst scoring Subfamily 2 member (5e-34) and the best scoring Subfamily 1 member (4.3e-07). This analysis shows that the Profile HMM prepared for Subfamily 2 phosphate acetyltransferases (PTA) distinguishes PTA sequences from phosphate butyryltransferase PTB protein sequences.
(170) Based on these analyses, 361 phosphate acetyltransferase sequences (PTA_PTB domain only) were identified and are provided in Table 12a.
(171) TABLE-US-00008 TABLE 12a SEQ ID NOs of phosphotransacetylase target gene coding regions and proteins GENBANK Nucleic GENBANK Amino Nucleotide Acid Amino Acid Acid GI Sequence SEQ Sequence SEQ Source Number Accession Information ID NO: Accession No. ID NO: Organism 255994631 complement 762 ZP_05427766.1 1123 Eubacterium (NZ_ACON01000003.1: saphenum 407639 . . . 408607) ATCC 49989 223935781 NZ_ABOX02000007.1: 763 ZP_03627696.1 1124 bacterium 10458 . . . 11600 Ellin514 196232920 NZ_ABVL01000018.1: 764 ZP_03131770.1 1125 Chthoniobacter 134406 . . . 135449 flavus Ellin428 187735919 NC_010655.1: 765 YP_001878031.1 1126 Akkermansia 1714393 . . . 1715484 muciniphila ATCC BAA-835 237732443 complement 766 ZP_04562924.1 1127 Citrobacter (NZ_GG657366.1: sp. 30_2 2437071 . . . 2438087) 157144617 NC_009792.1: 767 YP_001451936.1 1128 Citrobacter 326993 . . .328009 koseri ATCC BAA-895 56412650 NC_006511.1: 768 YP_149725.1 1129 Salmonella 473496 . . . 474512 enterica subsp. enterica serovar Paratyphi A str. ATCC 9150 161502384 NC_010067.1: 769 YP_001569496.1 1130 Salmonella 425125 . . . 426141 enterica subsp. arizonae serovar 62:z4, z23:-- 16130383 complement 770 NP_416953.1 1131 Escherichia (NC_000913.2: coli str. 2570511 . . . 571527) K-12 substr. MG1655 238895918 complement 771 YP_002920654.1 1132 Klebsiella (NC_012731.1: pneumoniae 3863205 . . . 3864221) NTUH-K2044 238794182 complement 772 ZP_04637797.1 1133 Yersinia (NZ_AALF02000025.1: intermedia 17041 . . . 18039) ATCC 29909 90414632 complement 773 ZP_01222604.1 1134 Photobacterium (NZ_AAPH01000046.1: profundum 12550 . . . 13527) 3TCK 163749608 complement 774 ZP_02156855.1 1135 Shewanella (NZ_ABIC01000008.1: benthica 73942 . . . 74913) KT99 120554157 NC_008740.1: 775 YP_958508.1 1136 Marinobacter 1389827 . . . 1390810 aquaeolei VT8 51246887 complement 776 YP_066771.1 1137 Desulfotalea (NC_006138.1: psychrophila 3433697 . . . 3434677) LSv54 226362753 complement 777 YP_002780531.1 1138 Rhodococcus (NC_012522.1: opacus 3613049 . . . 3614074) B4 111020534 complement 778 YP_703506.1 1139 Rhodococcus (NC_008268.1: jostii 3751557 . . . 3752585) RHA1 256669010 NZ_ACEV01000044.1: 779 ZP_05479963.1 1140 Streptomyces 80096 . . . 81085 sp. AA4 226227292 complement 780 YP_002761398.1 1141 Gemmatimonas (NC_012489.1: aurantiaca 2203452 . . . 2204474) T-27 239627158 complement 781 ZP_04670189.1 1142 Clostridiales (NZ_DS990260.1: bacterium 924717 . . . 925736) 1_7_47FAA 256753163 complement 782 ZP_05493958.1 1143 Clostridium (NZ_ACXX01000001.1: papyrosolvens 167634 . . . 168635) DSM 2782 257063834 NC_013165.1: 783 YP_003143506.1 1144 Slackia 1278820 . . 1279818 heliotrinireducens DSM 20476 254477436 NZ_DS999054.1: 784 ZP_05090822.1 1145 Ruegeria 3384063 . . . 3384914 sp. R11 126732220 complement 785 ZP_01748021.1 1146 Sagittula (NZ_AAYA01000016.1: stellata 68052 . . . 68996) E-37 19704507 complement 786 NP_604069.1 1147 Fusobacterium (NC_003454.1: nucleatum subsp. 1833702 . . . 1834715) nucleatum ATCC 25586 260494604 NZ_GG704456.1: 787 ZP_05814734.1 1148 Fusobacterium 222376 . . . 223389 sp. 3_1_33 262067001 complement 788 ZP_06026613.1 1149 Fusobacterium (NZ_ACJY01000064.1: periodonticum 9865 . . . 10869) ATCC 33693 257452333 complement 789 ZP_05617632.1 1150 Fusobacterium (NZ_ACDD01000037.1: sp. 3_1_5R 1250 . . . 2263) 257463639 NZ_ACDG01000104.1: 790 ZP_05628030.1 1151 Fusobacterium 17109 . . . 18122 sp. D12 253583748 complement 791 ZP_04860946.1 1152 Fusobacterium (NZ_GG696122.1: varium 645905 . . . 646912) ATCC 27725 237736963 NZ_GG657909.1: 792 ZP_04567444.1 1153 Fusobacterium 489336 . . . 490343 mortiferum ATCC 9817 157736754 complement 793 YP_001489437.1 1154 Arcobacter (NC_009850.1: butzleri 500921 . . . 501916) RM4018 257125122 NC_013192.1: 794 YP_003163236.1 1155 Leptotrichia 327731 . . . 328735 buccalis C-1013-b 260891157 NZ_ACVB02000026.1: 795 ZP_05902420.1 1156 Leptotrichia 170989 . . . 171993 hofstadii F0254 262037878 complement 796 ZP_06011308.1 1157 Leptotrichia (NZ_ADAD01000064.1: goodfellowii 9188 . . . 10195) F0264 229859891 NZ_ABUT01000004.1: 797 ZP_04479548.1 1158 Streptobacillus 63190 . . . 64215 moniliformis DSM 12112 227371784 NZ_ABVB01000002.1: 798 ZP_03855267.1 1159 Veillonella 260421 . . . 261419 parvula DSM 2008 227498373 complement 799 ZP_03928523.1 1160 Acidaminococcus (NZ_ACGB01000001.1: sp. D21 109630 . . . 110643) 42525561 NC_002967.9: 800 NP_970659.1 1161 Treponema 48816 . . . 49823 denticola ATCC 35405 257456313 NZ_ACYH01000011.1: 801 ZP_05621510.1 1162 Treponema 210840 . . . 211847 vincentii ATCC 35580 15639088 NC_000919.1: 802 NP_218534.1 1163 Treponema 102879 . . . 103889 pallidum subsp. pallidum str. Nichols 228000316 complement 803 ZP_04047318.1 1164 Brachyspira (NZ_ABTG01000001.1: murdochii 1179041 . . . 1180048) DSM 12563 225619252 NC_012225.1: 804 YP_002720478.1 1165 Brachyspira 340269 . . . 341276 hyodysenteriae WA1 218960931 complement 805 YP_001740706.1 1166 Candidatus (NS_000195.1: Cloacamonas 716420. . . 717424) acidaminovorans 239878221 complement 806 EER05013.1 1167 Perkinsus (join(GG681098.1: marinus 49679 . . . 49966, ATCC 50983 GG681098.1: 50017 . . . 50325, GG681098.1: 50380 . . . 50442, GG681098.1: 50494 . . . 50605, GG681098.1: 50656 . . . 50780, GG681098.1: 50826 . . . 50908, GG681098.1: 50958 . . . 51039)) 119953373 NC_008710.1: 807 YP_945582.1 1168 Borrelia 614125 . . . 615171 turicatae 91E135 187918450 NC_010673.1: 808 YP_001884013.1 1169 Borrelia 616784 . . . 617842 hermsii DAH 203284493 NC_011229.1: 809 YP_002222233.1 1170 Borrelia 622676 . . . 623746 duttonii Ly 224534734 complement 810 ZP_03675306.1 1171 Borrelia (NZ_ABKB02000009.1: spielmanii 27640 . . . 28677) A14S 216263399 NZ_ABCU02000001.1: 811 ZP_03435394.1 1172 Borrelia 172066 . . . 173103 afzelii ACA-1 219685198 NZ_ABPZ02000001.1: 812 ZP_03540018.1 1173 Borrelia 172004 . . . 173041 garinii Far04 224532296 NZ_ABCY02000001.1: 813 ZP_03672928.1 1174 Borrelia 609419 . . . 610456 valaisiana VS116 15594934 NC_001318.1: 814 NP_212723.1 1175 Borrelia 608020 . . . 609078 burgdorferi B31 189485346 NS_000191.1: 815 YP_001956287.1 1176 uncultured 518918 . . . 519919 Termite group 1 bacterium phylotype Rs-D17 42560817 NC_005364.2: 816 NP_975268.1 1177 Mycoplasma 308545 . . . 309513 mycoides subsp. mycoides SC str. PG1 83319483 NC_007633.1: 817 YP_424216.1 1178 Mycoplasma 277239 . . . 278207 capricolum subsp. capricolum ATCC 27343 50364858 NC_006055.1: 818 YP_053283.1 1179 Mesoplasma 58892 . . . 59860 florum L1 110005214 complement 819 CAK99540.1 1180 Spiroplasma (AM285317.1: citri 14153 . . . 15130) 12045155 complement 820 NP_072966.1 1181 Mycoplasma (NC_000908.2: genitalium 368733 . . . 369695) G37 13508167 complement 821 NP_110116.1 1182 Mycoplasma (NC_000912.1: pneumoniae 515605 . . . 516567) M129 31544825 complement 822 NP_853403.1 1183 Mycoplasma (NC_004829.1: gallisepticum 851083 . . . 852075) R 26553955 complement 823 NP_757889.1 1184 Mycoplasma (NC_004432.1: penetrans 640803 . . . 641777) HF-2 54020554 complement 824 YP_116016.1 1185 Mycoplasma (NC_006360.1: hyopneumoniae 638554 . . . 639507) 232 240047219 NC_012806.1: 825 YP_002960607.1 1186 Mycoplasma 88435 . . . 89406 conjunctivae 148377406 NC_009497.1: 826 YP_001256282.1 1187 Mycoplasma 159649 . . . 160605 agalactiae PG2 238809713 complement 827 BAH69503.1 1188 Mycoplasma (AP009608.1: fermentans 242111 . . . 243064) PG18 71894663 complement 828 YP_278771.1 1189 Mycoplasma (NC_007294.1: synoviae 757812 . . . 758771) 53 15828708 NC_002771.1: 829 NP_326068.1 1190 Mycoplasma 274992 . . . 275948 pulmonis UAB CTIP 47459003 NC_006908.1: 830 YP_015865.1 1191 Mycoplasma 230100 . . . 231068 mobile 163K 148377754 NC_009497.1: 831 YP_001256630.1 1192 Mycoplasma 572993 . . . 573967 agalactiae PG2 116515056 NC_008513.1: 832 YP_802685.1 1193 Buchnera 131608 . . . 132594 aphidicola str. Cc (Cinara cedri) 187934490 NC_010674.1: 833 YP_001885432.1 1194 Clostridium 1263289 . . . 1264287 botulinum B str. Eklund 17B 150016048 NC_009617.1: 834 YP_001308302.1 1195 Clostridium 1384403 . . . 1385404 beijerinckii NCIMB 8052 254519224 complement 835 ZP_05131280.1 1196 Clostridium (NZ_EQ999773.1: sp. 7_2_43FAA 2015491 . . . 2016492) 182417251 NZ_ABDT01000035.2: 836 ZP_02948604.1 1197 Clostridium 9769 . . . 10770 butyricum 5521 18310707 complement 837 NP_562641.1 1198 Clostridium (NC_003366.1: perfringens 2001712 . . . 2002719) str. 13 255524273 complement 838 ZP_05391232.1 1199 Clostridium (NZ_ACVI01000014.1: carboxidivorans 63543 . . . 64547) P7 153954015 NC_009706.1: 839 YP_001394780.1 1200 Clostridium 1428554 . . . 1429555 kluyveri DSM 555 187778946 NZ_ABKW02000004.1: 840 ZP_02995419.1 1201 Clostridium 733017 . . . 734015 sporogenes ATCC 15579 28210926 NC_004557.1: 841 NP_781870.1 1202 Clostridium 1326340 . . . 1327359 tetani E88 253681395 NZ_ACSJ01000007.1: 842 ZP_04862192.1 1203 Clostridium 344343 . . . 345338 botulinum D str. 1873 118444574 complement 843 YP_878298.1 1204 Clostridium (NC_008593.1: novyi 1416375 . . . 1417373) NT 242260238 NZ_ACPD01000011.1: 844 ZP_04804960.1 1205 Clostridium 83320 . . . 84318 cellulovorans 743B 15895019 NC_003030.1: 845 NP_348368.1 1206 Clostridium 1890289 . . . 1891290 acetobutylicum ATCC 824 169247670 EU313773.1: 846 ACA51668.1 1207 Thermoanaerobacterium 40 . . . 1026 saccharolyticum 255257449 NZ_ACVG01000034.1: 847 ZP_05336886.1 1208 Thermoanaerobacterium 8635 . . . 9621 thermosaccharolyticum DSM 571 20807926 complement 848 NP_623097.1 1209 Thermoanaerobacter (NC_003869.1: tengcongensis 1451520 . . . 1452515) MB4 167040369 complement 849 YP_001663354.1 1210 Thermoanaerobacter (NC_010320.1: sp. X514 1738259 . . . 1739257) 220931863 NC_011899.1: 850 YP_002508771.1 1211 Halothermothrix 1110901 . . . 1111899 orenii H 168 258514457 complement 851 YP_003190679.1 1212 Desulfotomaculum (NC_013216.1: acetoxidans 1194895 . . . 1195899) DSM 771 188586231 complement 852 YP_001917776.1 1213 Natranaerobius (NC_010718.1: thermophilus 1692944 . . . 1693942) JW/NM-WN-LF 78044760 complement 853 YP_360288.1 1214 Carboxydothermus (NC_007503.1: hydrogenoformans 1302969 . . . 1303973) Z-2901 262295620 complement 854 EEY83551.1 1215 Bacteroides (GG705150.1: sp. 2_1_33B 648642 . . .649655) 154494088 complement 855 ZP_02033408.1 1216 Parabacteroides (NZ_AAXE02000107.1: merdae 241237 . . . 242250) ATCC 43184 34540818 complement 856 NP_905297.1 1217 Porphyromonas (NC_002950.2: gingivalis 1149763 . . . 1150773) W83 228471187 complement 857 ZP_04056000.1 1218 Porphyromonas (NZ_ACLR01000214.1: uenonis 23231 . . . 24238) 60-3 229496164 NZ_ACNN01000020.1: 858 ZP_04389884.1 1219 Porphyromonas 205218 . . . 206225 endodontalis ATCC 35406 160887812 complement 859 ZP_02068815.1 1220 Bacteroides (NZ_AAYH02000031.1: uniformis 6367 . . . 7386) ATCC 8492 218131945 NZ_ABVO01000052.1: 860 ZP_03460749.1 1221 Bacteroides 25694 . . . 26710 eggerthii DSM 20697 224536405 complement 861 ZP_03676944.1 1222 Bacteroides (NZ_ACCH01000118.1: cellulosilyticus 1796 . . . 2812) DSM 14838 53711769 NC_006347.1: 862 YP_097761.1 1223 Bacteroides 557297 . . . 558316 fragilis YCH46 237715344 complement 863 ZP_04545825.1 1224 Bacteroides (NZ_EQ973249.1: sp. D1 217217 . . . 218236) 224025178 NZ_ACBW01000140.1: 864 ZP_03643544.1 1225 Bacteroides 3350 . . . 4369 coprophilus DSM 18228 198274546 NZ_ABQC02000011.1: 865 ZP_03207078.1 1226 Bacteroides 44269 . . . 45279 plebeius DSM 17135 150003111 NC_009614.1: 866 YP_001297855.1 1227 Bacteroides 740818 . . . 741831 vulgatus ATCC 8482 258649233 complement 867 ZP_05736702.1 1228 Prevotella (NZ_ACIJ02000031.1: tannerae 14596 . . . 15612) ATCC 51259 261881160 NZ_ACKS01000109.1: 868 ZP_06007587.1 1229 Prevotella 4227 . . . 5276 bergensis DSM 17361 260593477 NZ_ACVA01000073.1: 869 ZP_05858935.1 1230 Prevotella 31053 . . . 32099 veroralis F0319 260910323 complement 870 ZP_05916997.1 1231 Prevotella (NZ_ACZS01000043.1: sp. oral taxon 472 34220 . . . 35257) str. F0295 212550465 complement 871 YP_002308782.1 1232 Candidatus (NC_011565.1: Azobacteroides 126538 . . . 127539) pseudotrichonymphae genomovar. CFP2 114566305 NC_008346.1: 872 YP_753459.1 1233 Syntrophomonas 872558 . . . 873550 wolfei subsp. wolfei str. Goettingen 139437229 NZ_AAVN02000001.1: 873 ZP_01771389.1 1234 Collinsella 368246 . . . 369226 aerofaciens ATCC 25986 210631306 complement 874 ZP_03296849.1 1235 Collinsella (NZ_ABXJ01000041.1: stercoris 3328 . . . 4308) DSM 13279 229814970 complement 875 ZP_04445308.1 1236 Collinsella (NZ_ABXH02000002.1: intestinalis 65772 . . . 66770) DSM 13280 221194458 complement 876 ZP_03567515.1 1237 Atopobium (NZ_ACFE01000001.1: rimae 86128 . . . 87273) ATCC 49626 257784450 complement 877 YP_003179667.1 1238 Atopobium (NC_013203.1: parvulum 723329 . . . 724309) DSM 20469 227516084 complement 878 ZP_03946133.1 1239 Atopobium (NZ_ACGK01000007.1: vaginae 63717 . . . 64691 DSM 15829 227872296 NZ_ACKX01000061.1: 879 ZP_03990654.1 1240 Oribacterium 10209 . . . 11261 sinus F0268 229824780 NZ_ACIN02000002.1: 880 ZP_04450849.1 1241 Abiotrophia 126870 . . . 127931 defectiva ATCC 49176 260443831 NZ_ACIQ01000073.1: 881 ZP_05797601.1 1242 Oribacterium 32192 . . . 33196 sp. oral taxon 078 str. F0262 225176688 complement 882 ZP_03730247.1 1243 Clostridium (NZ_ACFX01000006.1: sp. M62/1 113088 . . . 114089) 253578981 complement 883 ZP_04856252.1 1244 Ruminococcus (NZ_GG696046.1: sp. 5_1_39BFAA 364564 . . . 365595) 153813664 NZ_AAVO02000036.1: 884 ZP_01966332.1 1245 Ruminococcus 4823 . . . 5992 obeum ATCC 29174 255281061 complement 885 ZP_05345616.1 1246 Bryantella (NZ_ACCL02000005.1: formatexigens 162813 . . . 163811) DSM 14469 225571965 NZ_ACBZ01000008.1: 886 ZP_03780829.1 1247 Blautia 1408 . . . 2442 hydrogenotrophica DSM 10507 210612569 NZ_ABWO01000095.2: 887 ZP_03289360.1 1248 Clostridium 3132 . . . 4127 nexile DSM 1787 154505354 complement 888 ZP_02042092.1 1249 Ruminococcus (NZ_AAYG02000022.1: gnavus 50151 . . . 51146) ATCC 29149 197303064 NZ_ABOU02000039.1: 889 ZP_03168112.1 1250 Ruminococcus 71843 . . . 72838 lactaris ATCC 29176 153816169 complement 890 ZP_01968837.1 1251 Ruminococcus (NZ_AAVP02000015.1: torques 36559 . . . 37554) ATCC 27756 167758299 complement 891 ZP_02430426.1 1252 Clostridium (NZ_ABFY02000009.1: scindens 238358 . . . 239380) ATCC 35704 225570721 NZ_ABYI02000032.1: 892 ZP_03779744.1 1253 Clostridium 3477 . . . 4499 hylemonae DSM 15053 166031766 NZ_AAXA02000013.1: 893 ZP_02234595.1 1254 Dorea 54410 . . . 55414 formicigenerans ATCC 27755 153853264 complement 894 ZP_01994673.1 1255 Dorea (NZ_AAXB02000002.1: longicatena 216862 . . . 217857) DSM 13814 160879474 NC_010001.1: 895 YP_01558442.1 1256 Clostridium 1657582 . . . 1658577 phytofermentans ISDg 239624054 complement 896 ZP_04667085.1 1257 Clostridiales (NZ_DS990263.1: bacterium 658578 . . . 659573) 1_7_47FAA 160938034 complement 897 ZP_02085391.1 1258 Clostridium (NZ_ABCC02000027.1: bolteae 52316 . . . 53311) ATCC BAA-613 260437037 complement 898 ZP_05790853.1 1259 Butyrivibrio (NZ_ABWN01000017.1: crossotus 33488 . . . 34483) DSM 2876 154483586 complement 899 ZP_02026034.1 1260 Eubacterium (NZ_AAVL02000033.1: ventriosum 74910 . . . 75941) ATCC 27560 238916996 complement 900 YP_002930513.1 1261 Eubacterium (NC_012778.1: eligens 1076225 . . . 1077220) ATCC 27750 242309058 NZ_DS990446.1: 901 ZP_04808213.1 1262 Helicobacter 108718 . . . 109737 pullorum MIT 98-5489 224418114 complement 902 ZP_03656120.1 1263 Helicobacter (NZ_ABQS01000024.1: canadensis 18744 . . . 19745) MIT 98-5491 237752737 NZ_GG661974.1: 903 ZP_04583217.1 1264 Helicobacter 463241 . . . 464236 winghamensis ATCC BAA-430 32266808 complement 904 NP_860840.1 1265 Helicobacter (NC_004917.1: hepaticus 1266998 . . . 1267993) ATCC 51449 224436915 complement 905 ZP_03657896.1 1266 Helicobacter (NZ_ABQT01000013.1: cinaedi 10506 . . . 11522) CCUG 18818 167745652 complement 906 ZP_02417779.1 1267 Anaerostipes (NZ_ABAX03000002.1: caccae 101957 . . . 102961) DSM 14662 167765558 complement 907 ZP_02437622.1 1268 Clostridium (NZ_ABGC03000004.1: sp. SS2/1 50807 . . . 51820) 163814038 NZ_ABEY02000003.1: 908 ZP_02205430.1 1269 Coprococcus 15727 . . . 16794 eutactus ATCC 27759 168334441 complement 909 ZP_02692616.1 1270 Epulopiscium (NZ_ABEQ01000029.2: sp. N.t. 21420 . . . 22418) morphotype B 257791476 NC_013204.1: 910 YP_003182082.1 1271 Eggerthella 2035882 . . . 2036880 lenta DSM 2243 256827068 complement 911 YP_003151027.1 1272 Cryptobacterium (NC_013170.1: curtum 735166 . . . 736167) DSM 15641 257063929 complement 912 YP_003143601.1 1273 Slackia (NC_013165.1: heliotrinireducens 1407526 . . . 1408521) DSM 20476 256757417 complement 913 ZP_05498135.1 1274 Clostridium (NZ_ACXX01000078.1: papyrosolvens 7706 . . . 8701) DSM 2782 196254011 NZ_ABVG01000076.1: 914 ZP_03152606.1 1275 Clostridium 9016 . . . 10092 thermocellum JW20 146297046 complement 915 YP_001180817.1 1276 Caldicellulosiruptor (NC_009437.1: saccharolyticus 2185773 . . . 2186804) DSM 8903 349833 L23147.1: 916 AAA72041.1 1277 Methanosarcina 207 . . . 1208 thermophila 20092407 complement 917 NP_618482.1 1278 Methanosarcina (NC_003552.1: acetivorans 4448053 . . . 4449054) C2A 73669327 complement 918 YP_305342.1 1279 Methanosarcina (NC_007355.1: barkeri 2275987 . . . 2276988) str. Fusaro 163734840 NZ_ABIG01000010.1: 919 ZP_02142278.1 1280 Roseobacter 132890 . . . 133867 litoralis Och 149 110678177 complement 920 YP_681184.1 1281 Roseobacter (NC_008209.1: denitrificans 803709 . . . 804707) OCh 114 159044374 complement 921 YP_001533168.1 1282 Dinoroseobacter (NC_009952.1: shibae 1904769 . . . 1905794) DFL 12 254512869 NZ_DS999532.1: 922 ZP_05124935.1 1283 Rhodobacteraceae 435221 . . . 436255 bacterium KLH11 260432366 complement 923 ZP_05786337.1 1284 Silicibacter (NZ_GG704596.1: lacuscaerulensis 1443685 . . . 1444707) ITI-1157 150376990 NC_009620.1: 924 YP_001313586.1 1285 Sinorhizobium 1371590 . . . 1372624 medicae WSM419 16264720 NC_003078.1: 925 NP_437512.1 1286 Sinorhizobium 1058537 . . . 1059529 meliloti 1021 239833801 complement 926 ZP_04682129.1 1287 Ochrobactrum (NZ_ACQA01000002.1: intermedium 595918 . . . 596886) LMG 3301 153010822 complement 927 YP_001372036.1 1288 Ochrobactrum (NC_00968.1: anthropi 862920 . . . 863897) ATCC 49188 187919084 complement 928 YP_001888115.1 1289 Burkholderia (NC_010676.1: phytofirmans 423371 . . . 424405) PsJN 91779405 NC_007952.1: 929 YP_554613.1 1290 Burkholderia 2605754 . . . 2606791 xenovorans LB400 186470979 NC_010625.1: 930 YP_001862297.1 1291 Burkholderia 763673 . . . 764704 phymatum STM815 73537607 complement 931 YP_297974.1 1292 Ralstonia (NC_007348.1: eutropha 372720 . . . 373757) JMP134 194292312 NC_010530.1: 932 YP_002008219.1 1293 Cupriavidus 1692071 . . . 1693105 taiwanensis 161521061 NC_010086.1: 933 YP_001584488.1 1294 Burkholderia 1600765 . . . 1601856 multivorans ATCC 17616 206563034 complement 934 YP_002233797.1 1295 Burkholderia (NC_011001.1: cenocepacia 1288493 . . . 1289527) J2315 90412230 complement 935 ZP_01220235.1 1296 Photobacterium (NZ_AAPH01000013.1: profundum 616 . . . 1653) 3TCK 224825256 complement 936 ZP_03698361.1 1297 Lutiella (NZ_ACIS01000005.1: nitroferrum 7593 . . . 8612) 2002 148973982 complement 937 ZP_01811515.1 1298 Vibrionales (NZ_AAZW01000001.1: bacterium 115872 . . . 116945) SWAT-3 84385317 complement 938 ZP_00988349.1 1299 Vibrio (NZ_AAMR01000001.1: splendidus 227808 . . . 228881) 12B01 149187938 NZ_ABCH01000003.1: 939 ZP_01866234.1 1300 Vibrio 163877 . . . 164926 shilonii AK1 260776268 complement 940 ZP_05885163.1 1301 Vibrio (NZ_ACZN01000015.1: coralliilyticus 316917 . . . 317966) ATCC BAA-450 45862014 AY498613.1: 941 AAS78789.1 1302 Paracoccus 8897 . . . 9853 denitrificans 77404622 NC_007488.1: 942 YP_345196.1 1303 Rhodobacter 38175 . . . 39173 sphaeroides 2.4.1 23630309 AY134843.1: 943 AAN08490.1 1304 Castellaniella 2180 . . . 3148 defragrans 83952615 NZ_AALY01000004.1: 944 ZP_00961345.1 1305 Roseovarius 6833 . . . 7819 nubinhibens ISM 56698382 complement 945 YP_168755.1 1306 Ruegeria (NC_003911.11: pomeroyi 3772593 . . . 3773606) DSS-3 149912659 NZ_ABCR01000001.1: 946 ZP_01901193.1 1307 Roseobacter 262172 . . . 263161 sp. AzwK-3b 126736835 complement 947 ZP_01752570.1 1308 Roseobacter (NZ_AAYC01000001.1: sp. SK209-2-6 23010 . . . 24029) 163732628 complement 948 ZP_02140073.1 1309 Roseobacter (NZ_ABIG01000003.1: litoralis 152433 . . . 153374) Och 149 89055338 NC_007802.1: 949 YP_510789.1 1310 Jannaschia 2864232 . . . 2865239 sp. CCS1 254459737 NZ_DS995276.1: 950 ZP_05073153.1 1311 Rhodobacterales 713420 . . . 714472 bacterium HTCC2083 116620211 NC_008536.1: 951 YP_822367.1 1312 Candidatus 1336713 . . . 1337708 Solibacter usitatus Ellin6076 95930364 NZ_AAEW02000012.1: 952 ZP_01313101.1 1313 Desulfuromonas 74280 . . . 75281 acetoxidans DSM 684 77920135 NC_007498.2: 953 YP_357950.1 1314 Pelobacter 2984046 . . . 2985047 carbinolicus DSM 2380 222054722 complement 954 YP_002537084.1 1315 Geobacter (NC_011979.1: sp. FRC-32 1793784 . . . 1794785) 148265418 NC_009483.1: 955 YP_001232124.1 1316 Geobacter 3992460 . . . 2993461 uraniireducens Rf4 39997800 NC_002939.4: 956 NP_953751.1 1317 Geobacter 2984470 . . . 2985471 sulfurreducens PCA 78222253 complement 957 YP_384000.1 1318 Geobacter (NC_007517.1: metallireducens 1150703 . . . 1151704) GS-15 118579718 complement 958 YP_900968.1 1319 Pelobacter (NC_008609.1: propionicus 1359173 . . . 1360177) DSM 2379 189424275 complement 959 YP_001951452.1 1320 Geobacter (NC_010814.1: lovleyi 1256052 . . . 1257053) SZ 255059775 NZ_ACPJ01000030.1: 960 ZP_05311922.1 1321 Geobacter 50425 . . . 51426 sp. M18 253700569 complement 961 YP_003021758.1 1322 Geobacter (NC_012918.1: sp. M21 2257017 . . . 2258018) 77920440 complement 962 YP_358255.1 1323 Pelobacter (NC_007498.2: carbinolicus 3332135 . . . 3333136) DSM 2380 227423754 NZ_ABTN01000011.1: 963 ZP_03906856.1 1324 Denitrovibrio 41883 . . . 42872 acetiphilus DSM 12809 193215894 NC_011026.1: 964 YP_001997093.1 1325 Chloroherpeton 2629909 . . . 2630916 thalassium ATCC 35110 150386298 complement 965 ZP_01924858.1 1326 Victivallis (NZ_ABDE01000122.1: vadensis 6609 . . . 7619) ATCC BAA-548 217034411 NZ_ABSX01000018.1: 966 ZP_03439825.1 1327 Helicobacter 29972 . . . 31570 pylori 98-10 254779508 complement 967 YP_003057614.1 1328 Helicobacter (NC_012973.1: pylori 876982 . . . 878538) B38 188527730 complement 968 YP_001910417.1 1329 Helicobacter (NC_010698.2: pylori 936737 . . . 938293) Shi470 15611908 complement 969 NP_223559.1 1330 Helicobacter (NC_000921.1: pylori 920263 . . . 921822) J99 109947805 complement 970 YP_665033.1 1331 Helicobacter (NC_008229.1: acinonychis 1105678 . . . 1107201) str. Sheeba 148926656 NZ_AASY01000008.1: 971 ZP_01810337.1 1332 Campylobacter 52243 . . . 53778 jejuni subsp. jejuni CG8486 57167700 complement 972 ZP_00366840.1 1333 Campylobacter (NZ_AAFL01000001.1: coli 232544 . . . 234046) RM2228 57242590 complement 973 ZP_00370527.1 1334 Campylobacter (NZ_AAFJ01000004.1: upsaliensis 6136 . . . 7638) RM3195 222823645 NC_012039.1: 974 YP_002575219.1 1335 Campylobacter 612447 . . . 613916 lari RM2100 154148075 complement 975 YP_001406718.1 1336 Campylobacter (NC_009714.1: hominis 1098161 . . . 1099597) ATCC BAA-381 257459711 NZ_ACYG01000019.1: 976 ZP_05624820.1 1337 Campylobacter 245785 . . . 247383 gracilis RM3268 118475502 complement 977 YP_891988.1 1338 Campylobacter (NC_008599.1: fetus subsp. 824533 . . . 825927) fetus 82-40 157164211 NC_009802.1: 978 YP_001466901.1 1339 Campylobacter 1056736 . . . 1058103 concisus 13826 154173700 complement 979 YP_001408221.1 1340 Campylobacter (NC_009715.1: curvus 947742 . . . 949112) 525.92 255322202 NZ_ACVQ01000017.1: 980 ZP_05363348.1 1341 Campylobacter 43696 . . . 45081 showae RM3277 225351910 NZ_ABXX02000003.1: 981 ZP_03742933.1 1342 Bifidobacterium 117844 . . . 119514 pseudocatenulatum DSM 20438 171743080 complement 982 ZP_02918887.1 1343 Bifidobacterium (NZ_ABIX02000002.1: dentium 2348529 . . . 2350229) ATCC 27678 154487476 complement 983 ZP_02028883.1 1344 Bifidobacterium (NZ_AAX1302000028.1: adolescentis 209751 . . . 211442) L2-32 229817818 complement 984 ZP_04448100.1 1345 Bifidobacterium (NZ_ABYS02000004.1: angulatum 899528 . . . 901234) DSM 20098 223467350 complement 985 ZP_03618886.1 1346 Bifidobacterium (NZ_ACCG01000014.1: breve 93417 . . . 95159) DSM 20213 227546035 NZ_ACHI01000009.1: 986 ZP_03976084.1 1347 Bifidobacterium 13043 . . . 14755 longum subsp. infantis ATCC 55813 213692597 NC_011593.1: 987 YP_002323183.1 1348 Bifidobacterium 1898224 . . . 1899936 longum subsp. infantis ATCC 15697 224282865 complement 988 ZP_03646187.1 1349 Bifidobacterium (NZ_ABQP01000009.1: bifidum 208984 . . . 210654) NCIMB 41171 227507562 NZ_ACGF01000124.1: 989 ZP_03937611.1 1350 Gardnerella 44384 . . . 46048 vaginalis ATCC 14019 183601499 complement 990 ZP_02962869.1 1351 Bifidobacterium (NZ_ABOT01000001.1: animalis subsp. 192971 . . . 194653) lactis HN019 261337301 complement 991 ZP_05965185.1 1352 Bifidobacterium (NZ_ABXB03000001.1: gallicum 137782 . . . 139455) DSM 20093 154507766 NZ_AAYI02000004.1: 992 ZP_02043408.1 1353 Actinomyces 231567 . . . 233045 odontolyticus ATCC 17982 227494860 complement 993 ZP_03925176.1 1354 Actinomyces (NZ_ACFG01000030.1: coleocanis 86700 . . . 88295) DSM 15436 19553946 complement 994 NP_601948.1 1355 Corynebacterium (NC_003450.3: glutamicum 2936506 . . . 2937891) ATCC 13032 25029147 complement 995 NP_739201.1 1356 Corynebacterium (NC_004369.1: efficiens 2758982 . . . 2760496) YS-314 38234612 complement 996 NP_940379.1 1357 Corynebacterium (NC_002935.2: diphtheriae 2103677 . . . 2105128) NCTC 13129 252124104 complement 997 ZP_04835255.1 1358 Corynebacterium (NZ_ACSH01000003.1: matruchotii 61905 . . . 63305) ATCC 14266 227489285 NZ_ABYP01000094.1: 998 ZP_03919601.1 1359 Corynebacterium 61648 . . . 63048 glucuronolyticum ATCC 51867 258561950 NZ_ACLJ01000070.1: 999 ZP_05708623.1 1360 Corynebacterium 13162 . . . 14523 genitalium ATCC 33030 227547861 NZ_ACHJ01000017.1: 1000 ZP_03977910.1 1361 Corynebacterium 9541 . . . 10899 lipophiloflavum DSM 44291 227502015 NZ_ACGD01000004.1: 1001 ZP_03932064.1 1362 Corynebacterium 82938 . . . 84305 accolens ATCC 49725 255325798 NZ_ACVP01000037.1: 1002 ZP_05366890.1 1363 Corynebacterium 5746 . . . 7107 tuberculostearicum SK141 227505901 NZ_ACGE01000122.1: 1003 ZP_03935950.1 1364 Corynebacterium 37468 . . . 38826 striatum ATCC 6940 227834110 complement 1004 YP_002835817.1 1365 Corynebacterum (NC_012590.1: aurimucosum 2492850 . . . 2494214) ATCC 700975 68535315 NC_007164.1: 1005 YP_250020.1 1366 Corynebacterium 307337 . . . 308848 jeikeium K411 172041418 complement 1006 YP_001801132.1 1367 Corynebacterium (NC_010545.1: urealyticum 2018026 . . . 2019369) DSM 7109 237786249 complement 1007 YP_002906954.1 1368 Corynebacterium (NC_012704.1: kroppenstedtii 1975731 . . . 1977287) DSM 44385 213965099 complement 1008 ZP_03393297.1 1369 Corynebacterium (NZ_ABZU01000003.1: amycolatum 128017 . . . 129543) SK46 225075788 complement 1009 ZP_03718987.1 1370 Neisseria (NZ_ACEN01000020.1: flavescens 5387 . . . 6889) NRL30031/H210 255067101 NZ_ACKO02000012.1: 1010 ZP_05318956.1 1371 Neisseria 64851 . . . 66353 sicca ATCC 29256 161869564 complement 1011 YP_001598731.1 1372 Neisseria (NC_010120.1: meningitidis 603202 . . . 604836) 053442 238022551 NZ_ACJW02000003.1: 1012 ZP_04602977.1 1373 Kingella 751672 . . . 753150 oralis ATCC 51147 83592714 complement 1013 YP_426466.1 1374 Rhodospirillum (NC_007643.1: rubrum 1625036 . . . 1626802) ATCC 11170 32490929 complement 1014 NP_871183.1 1375 Wigglesworthia (NC_004344.2: glossinidia 212680 . . . 214818) endosymbiont of Glossina brevipalpis 27904667 NC_004545.1: 1015 NP_777793.1 1376 Buchnera 186377 . . . 188524 aphidicola str. Bp (Baizongia pistaciae) 261415723 complement 1016 YP_003249406.1 1377 Fibrobacter (NC_013410.1: succinogenes subsp. 1639393 . . . 1640796) succinogenes S85 219556226 NZ_ABQH01000061.1: 1017 ZP_03535302.1 1378 Mycobacterium <3 . . . 1196 tuberculosis T17 228471665 complement 1018 ZP_04056438.1 1379 Capnocytophaga (NZ_ACLQ01000003.1: gingivalis 154599 . . . 155657) ATCC 33624 256370675 NC_013123.1: 1019 YP_003108500.1 1380 Candidatus 116952 . . . 117950 Sulcia muelleri SMDSEM 6685772 X89084.1: 1020 P77844 1381 Corynebacterium 1009 . . . 199 glutamicum 227876041 NZ_ACKW01000045.1: 1021 ZP_03994160.1 1382 Mobiluncus 33898 . . . 34887 mulieris ATCC 35243 227492324 NZ_ACCQ01000004.1: 1022 ZP_03922640.1 1383 Mobiluncus 214416 . . . 215417 curtisii ATCC 43063 225027017 NZ_ACEP01000064.1: 1023 ZP_03716209.1 1384 Eubacterium 10650 . . . 11609 halli DSM 3353 225028951 complement 1024 ZP_03718143.1 1385 Eubacterium (NZ_ACEP01000172.1: hallii 22364 . . . 23416) DSM 3353 257438679 complement 1025 ZP_05614434.1 1386 Faecalibacterium (NZ_ACOP02000029.1: prausnitzii 99 . . . 1124) A2-165 154496156 complement 1026 ZP_02034852.1 1387 Bacteroides (NZ_AAXG02000004.1: capillosus 103391 . . . 104380) ATCC 29799 225376322 NZ_ACFY01000086.1: 1027 ZP_03753543.1 1388 Roseburia 6940 . . . 7992 inulinivorans DSM 16841 257414121 complement 1028 ZP_04745275.2 1389 Roseburia (NZ_ABYJ02000202.1: intestinalis 41125 . . . 42165) L1-82 238923816 NC_012781.1: 1029 YP_002937332.1 1390 Eubacterium 1324280 . . . 1325263 rectale ATCC 33656 160893459 NZ_AAYW02000007.1: 1030 ZP_02074244.1 1391 Clostridium 63870 . . . 64919 sp. L2-50 229829305 complement 1031 ZP_04455374.1 1392 Shuttleworthia (NZ_ACIP02000002.1: satelles 495313 . . . 496299) DSM 14600 218282181 complement 1032 ZP_03488480.1 1393 Eubacterium (NZ_ABYT01000061.1: biforme 45 . . . 1016) DSM 3989 160916120 complement 1033 ZP_02078327.1 1394 Eubacterium (NZ_ABAW02000025.1: dolichum 71684 . . . 72694) DSM 3991 160915347 NZ_ABAW02000020.1: 1034 ZP_02077559.1 1395 Eubacterium 14646 . . . 15638 dolichum DSM 3991 212697404 NZ_ABXA01000047.1: 1035 ZP_03305532.1 1396 Anaerococcus 41715 . . . 42701 hydrogenalis DSM 7454 256545936 complement 1036 ZP_05473291.1 1397 Anaerococcus (NZ_ACXU01000022.1: vaginalis 22101 . . . 23087) ATCC 51170 227501001 NZ_ACGC01000115.1: 1037 ZP_03931050.1 1398 Anaerococcus 23122 . . . 24108 tetradius ATCC 35098 257067207 complement 1038 YP_003153463.1 1399 Anaerococcus (NC_013171.1: prevotii 1868769 . . . 1869752) DSM 20548 227485732 complement 1039 ZP_03916048.1 1400 Anaerococcus (NZ_ABYO01000196.1: lactolyticus 43814 . . . 44833) ATCC 51172 19746077 NC_003485.1: 1040 NP_607213.1 1401 Streptococcus 903788 . . . 904783 pyogenes MGAS8232 13622266 AE004092.1: 1041 AAK34003.1 1402 Streptococcus 923921 . . . 924916 pyogenes M1 GAS 222153008 NC_012004.1: 1042 YP_002562185.1 1403 Streptococcus 834034 . . . 835026 uberis 0140J 225868503 NC_012470.1: 1043 YP_002744451.1 1404 Streptococcus 1034662 . . . 1035663 equi subsp. Zooepidemicus 254997415 AP010655.1: 1044 BAH88016.1 1405 Streptococcus 1031526 . . . 1032521 mutans NN2025 171779341 NZ_ABJK02000020.1: 1045 ZP_02920305.1 1406 Streptococcus 38474 . . . 39472 infantarius subsp. infantarius ATCC BAA-102 76787123 complement 1046 YP_329798.1 1407 Streptococcus (NC_007432.1: agalactiae 1155758 . . . 1156750) A909 228477151 complement 1047 ZP_04061789.1 1408 Streptococcus (NZ_ACLO01000062.1: salivarius 54543 . . . 55526) SK126 55821439 complement 1048 YP_139881.1 1409 Streptococcus (NC_006448.1: thermophilus 1286014 . . . 1286997) LMG 18311 237650772 NZ_ABZC01000093.1: 1049 ZP_04525024.1 1410 Streptococcus 10653 . . . 11627 pneumoniae CCRI 1974 262282806 complement 1050 ZP_06060573.1 1411 Streptococcus (NZ_GG7049411.1: sp. 2_1_36FAA 11119 . . . 12096) 146318711 complement 1051 YP_001198423.1 1412 Streptococcus (NC_009442.1: suis 1032399 . . . 1033379) 05ZYH33 42518809 NC_005362.1: 1052 NP_964739.1 1413 Lactobacillus 788505 . . . 789482 johnsonii NCC 533 58337025 NC_006814.3: 1053 YP_193610.1 1414 Lactobacillus 698578 . . . 699567 acidophilus NCFM 227893214 NZ_ACGU01000037.1: 1054 ZP_04011019.1 1415 Lactobacillus 27358 . . . 28347 ultunensis DSM 16047 227877224 NZ_ACKR01000025.1: 1055 ZP_03995297.1 1416 Lactobacillus 14947 . . . 15936 crispatus JV-V01 260102516 NZ_ACLM01000112.1: 1056 ZP_05752753.1 1417 Lactobacillus 6924 . . . 7913 helveticus DSM 20075 227525975 NZ_ACGQ01000041.1: 1057 ZP_03956024.1 1418 Lactobacillus 59881 . . . 60858 jensenii JV-V16 228854857 complement 1058 ZP_04645187.1 1419 Lactobacillus (NZ_ACOY01000013.1: jensenii 251390 . . . 252367) 269-3 104773739 NC_008054.1: 1059 YP_618719.1 1420 Lactobacillus 547017 . . . 548006 delbrueckii subsp. bulgaricus ATCC 11842 259501464 NZ_ACLN01000013.1: 1060 ZP_05744366.1 1421 Lactobacillus 15438 . . . 16418 iners DSM 13335 16080818 complement 1061 NP_391646.1 1422 Bacillus (NC_000964.3: subtilis subsp. 3865355 . . . 3866326) subtilis str. 168 154687884 complement 1062 YP_001423045.1 1423 Bacillus (NC_009725.1: amyloliquefaciens 3590964 . . . 3591935) FZB42 52082282 complement 1063 YP_081073.1 1424 Bacillus (NC_006270.3: licheniformis 3821313 . . . 3822284) ATCC 14580 194016487 complement 1064 ZP_03055101.1 1425 Bacillus (NZ. ABRX01000004.1: pumilus 144981 . . . 145952) ATCC 7061 212640578 complement 1065 YP_002317098.1 1426 Anoxybacillus (NC_011567.1: flavithermus 2748264 . . . 2749247) WK1 239828646 complement 1066 YP_002951270.1 1427 Geobacillus (NC_012793.1: sp. WCH70 3393094 . . . 3394068) 138896990 complement 1067 YP_001127443.1 1428 Geobacillus (NC_009328.1: thermodenitrificans 3468960 . . . 3469934) NG80-2 56421950 complement 1068 YP_149268.1 1429 Geobacillus (NC_006510.1: kaustophilus 3456185 . . . 3457165) HTA426 149182788 NZ_ABCF01000043.1: 1069 ZP_01861251.1 1430 Bacillus 11583 . . . 12554 sp. SG-1 205375387 NZ_ABFU01000065.2: 1070 ZP_03228176.1 1431 Bacillus 12128 . . . 13099 coahuilensis m4-4 89101108 complement 1071 ZP_01173945.1 1432 Bacillus sp. (NZ_AAOX01000058.1: NRRL B-14911 10738 . . . 11715) 23100477 complement 1072 NP_693944.1 1433 Oceanobacillus (NC_004193.1: iheyensis 3134492 . . . 3135466) HTE831 229187615 complement 1073 ZP_04314753.1 1434 Bacillus (NZ_ACLU01000117.1: cereus 31676 . . . 32647) BGSC 6E1 46908338 complement 1074 YP_014727.1 1435 Listeria (NC_002973.6: monocytogenes 2171357 . . . 2172334) str. 4b F2365 229555968 NZ_ACCR01000020.1: 1075 ZP_04443757.1 1436 Listeria grayi 18426 . . . 19403 DSM 20601 15616385 complement 1076 NP_244690.1 1437 Bacillus (NC_002570.2: halodurans 3947889 . . . 3948881) C-125 56965668 complement 1077 YP_177402.1 1438 Bacillus (NC_006582.1: clausii 4069370 . . . 4070365) KSM-K16 229917170 complement 1078 YP_002885816.1 1439 Exiguobacterium (NC_012673.1: sp. AT1b 1410227 . . . 1411216) 172056261 NC_010556.1: 1079 YP_001812721.1 1440 Exiguobacterium 233988 . . . 234974 sibiricum 255-15 163762281 NZ_ABHZ01000002.1: 1080 ZP_02169346.1 1441 Bacillus 94480 . . . 95457 selenitireducens MLS10 242372812 NZ_ACJB01000048.1: 1081 ZP_04818386.1 1442 Staphylococcus 8122 . . . 9111 epidermidis M23864: W1 223042925 complement 1082 ZP_03612973.1 1443 Staphylococcus (NZ_ACFR01000002.1: capitis 330954 . . . 331943) SK14 239636796 complement 1083 ZP_04677798.1 1444 Staphylococcus (NZ_ACPZ01000027.:1 warneri 569915 . . . 570904) L37603 27467277 NC_004461.1: 1084 NP_763914.1 1445 Staphylococcus 356818 . . . 357807 epidermidis ATCC 12228 258422775 NZ_ACKI01000006.1: 1085 ZP_05685678.1 1446 Staphylococcus 980 . . . 1966 aureus A9635 70727403 complement 1086 YP_254319.1 1447 Staphylococcus (NC_007168.1: haemolyticus 2403280 . . . 2404269) JCSC1435 228475091 NZ_ACLP01000011.1: 1087 ZP_04059818.1 1448 Staphylococcus 16037 . . . 17026 hominis SK119 150011041 EF456699.1: 1088 ABR57177.1 1449 Staphylococcus 1 . . . 987 xylosus 73663433 complement 1089 YP_302214.1 1450 Staphylococcus (NC_007356.1: saprophyticus subsp. 2190871 . . . 2191857) saprophyticus ATCC 15305 224475734 NC_012121.1: 1090 YP_002633340.1 1451 Staphylococcus 250258 . . . 251247 carnosus subsp. carnosus TM300 222152076 complement 1091 YP_002561236.1 1452 Macrococcus (NC_011999.1: caseolyticus 1968130 . . . 1969119) JCSC5402 227514417 NZ_ACGI01000058.1: 1092 ZP_03944466.1 1453 Lactobacillus 71225 . . . 72199 fermentum ATCC 14931 256848058 complement 1093 ZP_05553502.1 1454 Lactobacillus (NZ_GG698804.1: coleohominis 125094 . . . 126059) 101-4-CHN 227529580 NZ_ACGV01000117.1: 1094 ZP_03959629.1 1455 Lactobacillus 1634 . . . 2608 vaginalis ATCC 49540 148543634 NC_009513.1: 1095 YP_001271004.1 1456 Lactobacillus 451991 . . . 452965 reuteri DSM 20016 259502766 NZ_ACLL01000024.1: 1096 ZP_05745668.1 1457 Lactobacillus 18101 . . . 19072 antri DSM 16041 116618560 complement 1097 YP_818931.1 1458 Leuconostoc (NC_008531.1: mesenteroides subsp. 1461235 . . . 1462215) mesenteroides ATCC 8293 170016912 NC_010471.1: 1098 YP_001727831.1 1459 Leuconostoc 577967 . . . 578959 citreum KM20 241894748 complement 1099 ZP_04782044.1 1460 Weissella (NZ_ACKU01000002.1: paramesenteroides 15496 . . . 16476) ATCC 33313 118587037 NZ_AAUV01000054.1: 1100 ZP_01544468.1 1461 Oenococcus 44038 . . . 45132 oeni ATCC BAA-1163 259046893 complement 1101 ZP_05737294.1 1462 Granulicatella (NZ_ACKZ01000012.:1 adiacens 86436 . . . 87425) ATCC 49175 260584167 complement 1102 ZP_05851915.1 1463 Granulicatella (NZ_GG703805.1: elegans 786281 . . . 787264) ATCC 700633 163789527 complement 1103 ZP_02183965.1 1464 Carnobacterium (NZ_ABHH01000002.1: sp. AT7 8081 . . . 9061) 257870102 NZ_GG670288.1: 1104 ZP_05649755.1 1465 Enterococcus 145742 . . . 146725 gallinarum EG2 227517869 NZ_ACGL01000051.1: 1105 ZP_03947918.1 1466 Enterococcus 2376 . . . 3401 faecalis TX0104 227552175 complement 1106 ZP_03982224.1 1467 Enterococcus (NZ.ACHL01000118.1: faecium 1216 . . . 2232) TX1330 81428954 complement 1107 YP_395954.1 1468 Lactobacillus (NC_007576.1: sakei subsp. 1313600 . . . 1314586) sakei 23K 229823693 NZ_ACIL02000007.1: 1108 ZP_04449762.1 1469 Catonella 6499 . . . 7482 morbi ATCC 51271 125623617 NC_009004.1: 1109 YP_001032100.1 1470 Lactococcus 752099 . . . 753079 lactis subsp. cremoris MG1363 116494500 NC_008526.1: 1110 YP_806234.1 1471 Lactobacillus 981403 . . . 982380 casei ATCC 334 28377658 NC_004567.1: 1111 NP_784550.1 1472 Lactobacillus 748192 . . . 749169 plantarum WCFS1 116333321 NC_008497.1: 1112 YP_794848.1 1473 Lactobacillus 702374 . . . 703348 brevis ATCC 367 227524782 complement 1113 ZP_03954831.1 1474 Lactobacillus (NZ_ACGP01000192.1: hilgardii 2135 . . . 3112) ATCC 8290 11862872 AB035800.1: 1114 BAB19267.1 1475 Lactobacillus 1006 . . . 1992 sanfranciscensis 227528239 NZ_ACGS01000093.1: 1115 ZP_03958288.1 1476 Lactobacillus 64718 . . . 65695 ruminis ATCC 25644 90962126 complement 1116 YP_536042.1 1477 Lactobacillus (NC_007929.1: salivarius 1183945 . . . 1184922) UCC118 259504733 NZ_ACLK01000016.1: 1117 ZP_05747635.1 1478 Erysipelothrix 55937 . . . 56917 rhusiopathiae ATCC 19414 116492140 NC_008525.1: 1118 YP_803875.1 1479 Pediococcus 385259 . . . 386230 pentosaceus ATCC 25745 160946581 NZ_ABEE02000016.1: 1119 ZP_02093784.1 1480 Parvimonas 72101 . . . 73072 micra ATCC 33270 169825312 complement 1120 YP_001692923.1 1481 Finegoldia (NC_010376.1: magna 1782855 . . . 1783826) ATCC 29328 229542439 NZ_AAWV02000001.1: 1121 ZP_04431499.1 1482 Bacillus 1452854 . . . 1453825 coagulans 36D1 241888505 NZ_ACDZ02000004.1: 1122 ZP_04775813.1 1483 Gemella 11622 . . . 12602 haemolysans ATCC 10379
(172) In addition, 201 phosphate acetyltransferase sequences that are characterized by two domains (DRTGG and PTA_PTB) are provided in Table 12b. MSA and phylogenetic analysis were performed as described above. Percent identity with respect to experimentally verified (or human curated) sequences is equal to or larger than 40, except for 4 sequences derived from plant organisms. Furthermore, hmmer search of the 201 sequences against the profile HMM of subfamily 2 (Table 14), clearly indicates that all Group 2 sequences belong to the PTA subfamily (least significant Evalue is 4.1e-93).
(173) TABLE-US-00009 TABLE 12b SEQ ID NOs of phosphotransacetylase target gene coding regions and proteins GENBANK Nucleic GENBANK Amino Nucleotide Acid Amino Acid Acid GI Sequence SEQ Sequence SEQ Source Number Accession Information ID NO: Accession No. ID NO: Organism 152964825 complement 1484 YP_001360609.1 1685 Kineococcus (NC_009664.2: radiotolerans 1430885 . . . 1432984) SRS30216 88800302 complement 1485 ZP_01115869.1 1686 Reinekea (NZ_AAOE01000024.1: blandensis 59450 . . . 61609) MED297 254786809 complement 1486 YP_003074238.1 1687 Teredinibacter (NC_012997.1: turnerae 3139764 . . . 3141917) T7901 120554060 complement 1487 YP_958411.1 1688 Marinobacter (NC_008740.1: aquaeolei 1283732 . . . 1285885) VT8 83647145 NC_007645.1: 1488 YP_435580.1 1689 Hahella 4579211 . . . 4581358 chejuensis KCTC 2396 146308660 NC_009439.1: 1489 YP_001189125.1 1690 Pseudomonas 4021959 . . . 4024052 mendocina ymp 116048757 NC_008463.1: 1490 YP_792443.1 1691 Pseudomonas 4740071 . . . 4742185 aeruginosa UCBPP-PA14 28868382 complement 1491 NP_791001.1 1692 Pseudomonas (NC_004578.1: syringae pv. 1283902 . . . 1285992) tomato str. DC3000 70728320 complement 1492 YP_258069.1 1693 Pseudomonas (NC_004129.6: fluorescens 1081214 . . . 1083313) Pf-5 104780139 complement 1493 YP_606637.1 1694 Pseudomonas (NC_008027.1: entomophila 952666 . . . 954756) L48 226945506 complement 1494 YP_002800579.1 1695 Azotobacter (NC_012560.1: vinelandii 3530138 . . . 3532276) DJ 146281510 complement 1495 YP_001171663.1 1696 Pseudomonas (NC_009434.1: stutzeri 1238536 . . . 1240632) A1501 30248315 complement 1496 NP_840385.1 1697 Nitrosomonas (NC_004757.1: europaea 326321 . . . 328408) ATCC 19718 226946148 NC_012560.1: 1497 YP_002801221.1 1698 Azotobacter 4145609 . . . 4147684 vinelandii DJ 226357371 NC_012526.1: 1498 YP_002787111.1 1699 Deinococcus 2779899 . . . 2782016 deserti VCD115 94984159 complement 1499 YP_603523.1 1700 Deinococcus (NC_008025.1: geothermalis 46701 . . . 48812) DSM 11300 15805114 complement 1500 NP_293799.1 1701 Deinococcus (NC_001263.1: radiodurans 69707 . . . 71875) R1 89899079 complement 1501 YP_521550.1 1702 Rhodoferax (NC_007908.1: ferrireducens 264127 . . . 266178) T118 90422592 NC_007925.1: 1502 YP_530962.1 1703 Rhodopseudomonas 1181422 . . . 1183572 palustris BisB18 90423512 NC_007925.1: 1503 YP_531882.1 1704 Rhodopseudomonas 2183340 . . . 2185475 palustris BisB18 115525859 NC_008435.1: 1504 YP_782770.1 1705 Rhodopseudomonas 4320999 . . . 4323140 palustris BisA53 167574473 complement 1505 ZP_02367347.1 1706 Burkholderia (NZ_ABBG01000507.1: oklahomensis 7891 . . . 9969) C6786 83594327 complement 1506 YP_428079.1 1707 Rhodospirillum (NC_007643.1: rubrum 3449832 . . . 3451943) ATCC 11170 90422165 NC_007925.1: 1507 YP_530535.1 1708 Rhodopseudomonas 696325 . . . 698388 palustris BisB18 34496985 complement 1508 NP_901200.1 1709 Chromobacterium (NC_005085.1: violaceum 1636285 . . . 1638366) ATCC 12472 224825239 complement 1509 ZP_03698345.1 1710 Lutiella (NZ_ACIS01000004.1: nitroferrum 398128 . . . 400215) 2002 148652157 complement 1510 YP_001279250.1 1711 Psychrobacter (NC_009524.1: sp. PRwf-1 415573 . . . 417720) 93005047 complement 1511 YP_579484.1 1712 Psychrobacter (NC_007969.1: cryohalolentis 257926 . . . 260082) K5 257453691 NZ_ACYI01000010.1: 1512 ZP_05618978.1 1713 Enhydrobacter 16653 . . . 18797 aerosaccus SK60 255321153 NZ_ACVR01000080.1: 1513 ZP_05362319.1 1714 Acinetobacter 44385 . . . 46529 radioresistens SK82 50083778 complement 1514 YP_045288.1 1715 Acinetobacter (NC_005966.1: sp. ADP1 527524 . . . 529686) 260549093 NZ_GG704496.1: 1515 ZP_05823314.1 1716 Acinetobacter 86045 . . . 88189 sp. RUH2624 226953952 complement 1516 ZP_03824416.1 1717 Acinetobacter (NZ_ABYN01000201.1: sp. ATCC 27244 23157 . . . 25289) 153005955 NC_009675.1: 1517 YP_001380280.1 1718 Anaeromyxobacter 3624676 . . . 3626811 sp. Fw109-5 86159318 complement 1518 YP_466103.1 1719 Anaeromyxobacter (NC_007760.1: dehalogenans 3267950 . . . 3270094) 2CP-C 52425053 complement 1519 YP_088190.1 1720 Mannheimia (NC_006300.1: succiniciproducens 977458 . . . 979596) MBEL55E 152979320 NC_009655.1: 1520 YP_001344949.1 1721 Actinobacillus 1823344 . . . 1825485 succinogenes 130Z 251792685 NC_012913.1: 1521 YP_003007411.1 1722 Aggregatibacter 968721 . . . 970856 aphrophilus NJ8700 145633066 NZ_AAZF01000004.1: 1522 ZP_01788798.1 1723 Haemophilus 73469 . . . 75604 influenzae 3655 113460945 complement 1523 YP_719012.1 1724 Haemophilus (NC_008309.1: somnus 873911 . . . 876049) 129PT 15602570 NC_002663.1: 1524 NP_245642.1 1725 Pasteurella 821181 . . . 823319 multocida subsp. multocida str. Pm70 260913970 complement 1525 ZP_05920444.1 1726 Pasteurella (NZ_ACZR01000013.1: dagmatis 172766 . . . 174904) ATCC 43325 53729159 complement 1526 ZP_00133992.2 1727 Actinobacillus (NZ_AACK01000004.1: pleuropneumoniae 12180 . . . 14318) serovar 1 str. 4074 240949203 NZ_ACQL01000097.1: 1527 ZP_04753547.1 1728 Actinobacillus 15931 . . . 18069 minor NM305 33152520 NC_002940.2: 1528 NP_873873.1 1729 Haemophilus 1192390 . . . 1194528 ducreyi 35000HP 254362832 NZ_DS264681.1: 1529 ZP_04978908.1 1730 Mannheimia 4949 . . . 7084 haemolytica PHL213 219870647 NC_011852.1: 1530 YP_002475022.1 1731 Haemophilus 435431 . . . 437566 parasuis SH0165 258637834 NZ_ACYJ01000022.1: 1531 ZP_05730581.1 1732 Pantoea 41620 . . . 43764 sp. At-9b 188533336 complement 1532 YP_001907133.1 1733 Erwinia (NC_010694.1: tasmaniensis 1324250 . . . 1326379) Et1/99 85059585 NC_007712.1: 1533 YP_455287.1 1734 Sodalis 2759501 . . . 2761645 glossinidius str. morsitans 258631105 complement 1534 ZP_05723922.1 1735 Dickeya (NZ_ACYK01000004.1: dadantii 104546 . . . 106687) Ech586 261820783 complement 1535 YP_003258889.1 1736 Pectobacterium (NC_013421.1: wasabiae 1606509 . . . 1608647) WPP163 242239978 NC_012880.1: 1536 YP_002988159.1 1737 Dickeya 3004583 . . . 3006724 dadantii Ech703 22125515 complement 1537 NP_668938.1 1738 Yersinia (NC_004088.1: pestis 1788905 . . . 1791058) KIM 10 157371554 NC_009832.1: 1538 YP_001479543.1 1739 Serratia 3673293 . . . 3675482 proteamaculans 568 238920583 NC_012779.1: 1539 YP_002934098.1 1740 Edwardsiella 2589897 . . . 2592035 ictaluri 93-146 197285630 NC_010554.1: 1540 YP_002151502.1 1741 Proteus 1898593 . . . 1900737 mirabilis HI4320 37526984 NC_005126.1: 1541 NP_930328.1 1742 Photorhabdus 3612456 . . . 3614597 luminescens subsp. laumondii TTO1 238895817 NC_012731.1: 1542 YP_002920553.1 1743 Klebsiella 3763302 . . . 3765449 pneumoniae NTUH-K2044 146312483 NC_009436.1: 1543 YP_001177557.1 1744 Enterobacter 3080629 . . . 3082770 sp. 638 260598715 NC_013282.1: 1544 YP_003211286.1 1745 Cronobacter 3057676 . . . 3059814 turicensis 601935 D21123.1: 1545 BAA04663.1 1746 Escherichia 77 . . . 2218 coli 238898722 complement 1546 YP_002924403.1 1747 Candidatus (NC_012751.1: Hamiltonella 1494526 . . . 1496655) defensa 5AT (Acyrthosiphon pisum) 227114079 NZ_ABVX01000029.1: 1547 ZP_03827735.1 1748 Pectobacterium 23117 . . . 25261 carotovorum subsp. brasiliensis PBR1692 89072717 complement 1548 ZP_01159282.1 1749 Photobacterium (NZ_AAOU01000004.1: sp. SKA34 98074 . . . 100221) 54309953 NC_006370.1: 1549 YP_130973.1 1750 Photobacterium 3245262 . . . 3247418 profundum SS9 262274670 NZ_ADAQ01000011.1: 1550 ZP_06052481.1 1751 Grimontia 496361 . . . 498520 hollisae CIP 101886 260768101 complement 1551 ZP_05877035.1 1752 Vibrio (NZ_ACZP01000013.1: furnissii 239301 . . . 241427) CIP 102972 260773044 NZ_ACZO01000006.1: 1552 ZP_05881960.1 1753 Vibrio 1066216 . . . 1068360 metschnikovii CIP 69.14 163802859 complement 1553 ZP_02196748.1 1754 Vibrio (NZ_ABGR01000013.1: sp. AND4 61871 . . . 64036) 37680318 NC_005139.1: 1554 NP_934927.1 1755 Vibrio 2144915 . . . 2147059 vulnificus YJ016 149188151 complement 1555 ZP_01866446.1 1756 Vibrio (NZ_ABCH01000004.1: shilonii 163878 . . . 166022) AK1 218708991 complement 1556 YP_002416612.1 1757 Vibrio (NC_011753.1: splendidus 1031606 . . . 1033810) LGP32 209695557 NC_011312.1: 1557 YP_002263486.1 1758 Aliivibrio 2262635 . . . 2264806 salmonicida LFI1238 229525709 complement 1558 ZP_04415114.1 1759 Vibrio (NZ_ACHV01000001.1: cholerae bv. 2574339 . . . 2576483) albensis VL426 145300284 NC_009348.1: 1559 YP_001143125.1 1760 Aeromonas 3681431 . . . 3683584 salmonicida subsp. salmonicida A449 237807651 complement 1560 YP_002892091.1 1761 Tolumonas (NC_012691.1: auensis 958413 . . . 960569) DSM 9187 90407162 complement 1561 ZP_01215350.1 1762 Psychromonas (NZ_AAPG01000006.1: sp. CNPT3 41954 . . . 44116) 119946918 NC_008709.1: 1562 YP_944598.1 1763 Psychromonas 4084304 . . . 4086466 ingrahamii 37 157374843 complement 1563 YP_001473443.1 1764 Shewanella (NC_009831.1: sediminis 2041698 . . . 2043839) HAW-EB3 170727231 NC_010506.1: 1564 YP_001761257.1 1765 Shewanella 3531467 . . . 3533608 woodyi ATCC 51908 127513322 NC_009092.1: 1565 YP_001094519.1 1766 Shewanella 2807561 . . . 2809699 loihica PV-4 167624517 NC_010334.1: 1566 YP_001674811.1 1767 Shewanella 3149368 . . . 3151515 halifaxensis HAW-EB4 117919999 complement 1567 YP_869191.1 1768 Shewanella (NC_008577.1: sp. ANA-3 1806421 . . . 180857 119774631 complement 1568 YP_927371.1 1769 Shewanella (NC_008700.1: amazonensis 1807689 . . . 1809827) SB2B 114563647 NC_008345.1: 1569 YP_751160.1 1770 Shewanella 2956515 . . . 2958662 frigidimarina NCIMB 400 91793762 NC_007954.1: 1570 YP_563413.1 1771 Shewanella 2868611 . . . 2870791 denitrificans OS217 157376672 NC_009831.1: 1571 YP_001475272.1 1772 Shewanella 4313346 . . . 4315484 sediminis HAW-EB3 167624655 complement 1572 YP_001674949.1 1773 Shewanella (NC_010334.1: halifaxensis 3320048 . . . 3322198) HAW-EB4 239996136 complement 1573 ZP_04716660.1 1774 Alteromonas (NZ_ABQB01000564.1: macleodii 6079 . . . 8301) ATCC 27126 109898905 NC_008228.1: 1574 YP_662160.1 1775 Pseudoalteromonas 3144369 . . . 3146513 atlantica T6c 119469286 NZ_AAVS01000006.1: 1575 ZP_01612225.1 1776 Alteromonadales 30053 . . . 32206 bacterium TW-7 88860001 complement 1576 ZP_01134640.1 1777 Pseudoalteromonas (NZ_AAOH01000005.1: tunicata 230650 . . . 232797) D2 71282469 NC_003910.7: 1577 YP_269873.1 1778 Colwellia 3309465 . . . 3311585 psychrerythraea 34H 152996332 NC_009654.1: 1578 YP_001341167.1 1779 Marinomonas 2608121 . . . 2610220 sp. MWYL1 87121463 NZ_AANE01000011.1: 1579 ZP_01077352.1 1780 Marinomonas 112021 . . . 114096 sp. MED121 146328905 NC_009446.1: 1580 YP_001209362.1 1781 Dichelobacter 489780 . . . 491837 nodosus VCS1703A 258544959 NZ_ACKY01000059.1: 1581 ZP_05705193.1 1782 Cardiobacterium 4448 . . . 6562 hominis ATCC 15826 262104765 complement 1582 EEY62817.1 1783 Phytophthora (DS028152.1: infestans 677306 . . . 679621) T30-4 262104764 complement 1583 EEY62816.1 1784 Phytophthora (DS028152.1: infestans 674496 . . . 676781) T30-4 159472743 XM_001694452.1: 1584 XP_001694504.1 1785 Chlamydomonas 258 . . . 2636 reinhardtii 168000833 XM_001753068.1: 1585 XP_001753120.1 1786 Physcomitrella 1 . . . 2367 patens subsp. Patens 172038009 complement 1586 YP_001804510.1 1787 Cyanothece (NC_010546.1: sp. ATCC 51142 3214848 . . . 3216944) 126658068 NZ_AAXW01000014.1: 1587 ZP_01729220.1 1788 Cyanothece 79066 . . . 81162 sp. CCY0110 257060449 NC_013161.1: 1588 YP_003138337.1 1789 Cyanothece 2659296 . . . 2661419 sp. PCC 8802 218441705 complement 1589 YP_002380034.1 1790 Cyanothece (NC_011729.1: sp. PCC 7424 5341705 . . . 5343810) 166368837 NC_010296.1: 1590 YP_001661110.1 1791 Microcystis 5646854 . . . 5648950 aeruginosa NIES-843 220909840 NC_011884.1: 1591 YP_002485151.1 1792 Cyanothece 4551169 . . . 4553265 sp. PCC 7425 16330299 NC_000911.1: 1592 NP_441027.1 1793 Synechocystis 1250442 . . . 1252535 sp. PCC 6803 86142732 NZ_AANC01000005.1: 1593 ZP_01061171.1 1794 Leeuwenhoekiella 209172 . . . 211268 blandensis MED217 146301271 complement 1594 YP_001195862.1 1795 Flavobacterium (NC_009441.1: johnsoniae 4208789 . . . 4210882) UW101 260061847 NC_013222.1: 1595 YP_003194927.1 1796 Robiginitalea 1408358 . . . 1410454 biformata HTCC2501 88713711 complement 1596 ZP_01107792.1 1797 Flavobacteriales (NZ_AAOC01000008.1: bacterium 22821 . . . 24917) HTCC2170 86133149 complement 1597 ZP_01051731.1 1798 Polaribacter (NZ_CH902588.1: sp. MED152 146636 . . . 148729) 88803680 NZ_AAOG01000005.1: 1598 ZP_01119204.1 1799 Polaribacter 54861 . . . 56951 irgensii 23-P 213962668 NZ_ABZV01000006.1: 1599 ZP_03390929.1 1800 Capnocytophaga 103363 . . . 105438 sputigena ATCC 33612 256820698 complement 1600 YP_003141977.1 1801 Capnocytophaga (NC_013162.1: ochracea 2243972 . . . 2246047) DSM 7271 46581432 NC_002937.3: 1601 YP_012240.1 1802 Desulfovibrio 3152216 . . . 3154330 vulgaris str. Hildenborough 218886955 NC_011769.1: 1602 YP_002436276.1 1803 Desulfovibrio 2286534 . . . 2288648 vulgaris str. Miyazaki F 78358281 NC_007519.1: 1603 YP_389730.1 1804 Desulfovibrio 3235663 . . . 3237822 desulfuricans subsp. desulfuricans str. G20 242280036 complement 1604 YP_002992165.1 1805 Desulfovibrio (NC_012881.1: salexigens 2812652 . . . 2814769) DSM 2638 258405159 complement 1605 YP_003197901.1 1806 Desulfohalobium (NC_013223.1: retbaense 1218708 . . . 1220816) DSM 5692 256828849 NC_013173.1: 1606 YP_003157577.1 1807 Desulfomicrobium 1143375 . . . 1145477 baculatum DSM 4028 225198782 complement 1607 ZP_03737911.1 1808 Desulfonatronospira (NZ_ACJN01000010.1: thiodismutans 60728 . . . 62824) ASO3-1 242278203 NC_012881.1: 1608 YP_002990332.1 1809 Desulfovibrio 802309 . . . 804414 salexigens DSM 2638 212704109 complement 1609 ZP_03312237.1 1810 Desulfovibrio (NZ_ABXU01000065.1: piger 34368 . . . 36470) ATCC 29098 220903578 complement 1610 YP_002478890.1 1811 Desulfovibrio (NC_011883.1: desulfuricans 357004 . . . 359112) subsp. desulfuricans str. ATCC 27774 51244410 NC_006138.1: 1611 YP_064294.1 1812 Desulfotalea 608983 . . . 611115 psychrophila LSv54 94986723 NC_008011.1: 1612 YP_594656.1 1813 Lawsonia 347892 . . . 350012 intracellularis PHE/MN1-00 119488858 complement 1613 ZP_01621820.1 1814 Lyngbya (NZ_AAVU01000021.1: sp. PCC 8106 33384 . . . 35414) 209524350 NZ_ABYK01000010.1: 1614 ZP_03272899.1 1815 Arthrospira 35808 . . . 37916 maxima CS-328 116748909 NC_008554.1: 1615 YP_845596.1 1816 Syntrophobacter 1826816 . . . 1828915 fumaroxidans MPOB 241776655 NZ_ACQQ01000008.1: 1616 ZP_04773932.1 1817 Allochromatium 86078 . . . 88198 vinosum DSM 180 32476008 NC_005027.1: 1617 NP_869002.1 1818 Rhodopirellula 5198833 . . . 5200932 baltica SH 1 78776256 NC_007575.1: 1618 YP_392571.1 1819 Sulfurimonas 60204 . . . 62282 denitrificans DSM 1251 254458291 complement 1619 ZP_05071717.1 1820 Campylobacterales (NZ_DS995288.1: bacterium 173480 . . . 175561) GD 1 229532518 NZ_ABUV01000006.1: 1620 ZP_04421899.1 1821 Sulfurospirillum 73569 . . . 75677 deleyianum DSM 6946 152993574 NC_009663.1: 1621 YP_001359295.1 1822 Sulfurovum 2069625 . . . 2071724 sp. NBC37-1 120401715 NC_008726.1: 1622 YP_951544.1 1823 Mycobacterium 740616 . . . 742694 vanbaalenii PYR-1 145220810 complement 1623 YP_001131488.1 1824 Mycobacterium (NC_009338.1: gilvum 189392 . . . 191515) PYR-GCK 108797517 NC_008146.1: 1624 YP_637714.1 1825 Mycobacterium 594117 . . . 596231 sp. MCS 118473540 NC_008596.1: 1625 YP_885188.1 1826 Mycobacterium 867578 . . . 869656 smegmatis str. MC2 155 169631304 complement 1626 YP_001704953.1 1827 Mycobacterium (NC_010397.1: abscessus 4294451 . . . 4296532) 240168870 NZ_ACBV01000011.1: 1627 ZP_04747529.1 1828 Mycobacterium 33884 . . . 35974 kansasii ATCC 12478 183980733 NC_010612.1: 1628 YP_001849024.1 1829 Mycobacterium 853987 . . . 856071 marinum M 15607549 NC_000962.2: 1629 NP_214922.1 1830 Mycobacterium 491786 . . . 493858 tuberculosis H37Rv 41409983 NC_002944.2: 1630 NP_962819.1 1831 Mycobacterium 4345845 . . . 4347932 avium subsp. paratuberculosis K-10 254818871 complement 1631 ZP_05223872.1 1832 Mycobacterium (NZ_ABIN01000026.1: intracellulare 2280 . . . 4439) ATCC 13950 226304961 NC_012490.1: 1632 YP_002764919.1 1833 Rhodococcus 1605870 . . . 1607948 erythropolis PR4 111019190 complement 1633 YP_702162.1 1834 Rhodococcus (NC_008268.1: jostii 2308925 . . . 2311045) RHA1 54027320 complement 1634 YP_121562.1 1835 Nocardia (NC_006361.1: farcinica 5652808 . . . 5654895) IFM 10152 227978095 NZ_ABVA01000001.1: 1635 ZP_04025361.1 1836 Tsukamurella 785441 . . . 787570 paurometabola DSM 20162 262204223 complement 1636 YP_003275431.1 1837 Gordonia (NC_013441.1: bronchialis 4704088 . . . 4706208) DSM 43247 256831883 complement 1637 YP_003160610.1 1838 Jonesia (NC_013174.1: denitrificans 683848 . . . 685947) DSM 20603 260517199 complement 1638 ZP_05816650.1 1839 Sanguibacter (NZ_ABUN01000002.1: keddieii 90744 . . . 92939) DSM 10542 229243856 complement 1639 ZP_04368027.1 1840 Cellulomonas (NZ_ABTJ01000131.1: flavigena 4381 . . . 6468) DSM 20109 229821528 NC_012669.1: 1640 YP_002883054.1 1841 Beutenbergia 3401218 . . . 3403323 cavernae DSM 12333 227428424 complement 1641 ZP_03911481.1 1842 Xylanimonas (NZ_ABVC01000008.1: cellulosilytica 150308 . . . 152407) DSM 15894 119717178 complement 1642 YP_924143.1 1843 Nocardioides (NC_008699.1: sp. JS614 3139954 . . . 3142044) 227381337 complement 1643 ZP_03864789.1 1844 Kribbella (NZ_ABUC01000011.1: flavida 233655 . . . 235784) DSM 17836 88856399 NZ_AAOB01000010.1: 1644 ZP_01131057.1 1845 marine 2970 . . . 5138 actinobacterium PHSC20C1 170780609 NC_010407.1: 1645 YP_001708941.1 1846 Clavibacter 179995 . . . 182112 michiganensis subsp. Sepedonicus 50954174 NC_006087.1: 1646 YP_061462.1 1847 Leifsonia 335128 . . . 337257 xyli subsp. xyli str. CTCB07 114331961 complement 1647 YP_748183.1 1848 Nitrosomonas (NC_008344.1: eutropha 2107910 . . . 2110039) C91 256395328 complement 1648 YP_003116892.1 1849 Catenulispora (NC_013131.1: acidiphila 7130890 . . . 7133034) DSM 44928 258650827 NC_013235.1: 1649 YP_003199983.1 1850 Nakamurella 639398 . . . 641491 multipartita DSM 44233 257068066 complement 1650 YP_003154321.1 1851 Brachybacterium (NC_013172.1: faecium 995592 . . . 997667) DSM 4810 227497260 complement 1651 ZP_03927492.1 1852 Actinomyces (NZ_ACFH01000109.1: urogenitalis 3054 . . . 5108) DSM 15434 256824971 NC_013169.1: 1652 YP_003148931.1 1853 Kytococcus 1149994 . . . 1152081 sedentarius DSM 20547 260455562 NZ_ACZH01000022.1: 1653 ZP_05803950.1 1854 Streptomyces 9845 . . . 11917 flavogriseus ATCC 33331 182435904 NC_010572.1: 1654 YP_001823623.1 1855 Streptomyces 2506931 . . . 2509012 griseus subsp. griseus NBRC 13350 254387454 NZ_DS570624.1: 1655 ZP_05002693.1 1856 Streptomyces 26512 . . . 28596 clavuligerus ATCC 27064 254400535 NZ_DS570905.1: 1656 ZP_05015493.1 1857 Streptomyces 178100 . . . 180199 sviceus ATCC 29083 256813645 NZ_ACFA01000303.1: 1657 ZP_05538660.1 1858 Streptomyces 9482 . . . 11584 griseoflavus Tu4000 239928836 NZ_ABYA01000185.1: 1658 ZP_04685789.1 1859 Streptomyces 2180 . . . 4282 ghanaensis ATCC 14672 256804684 complement 1659 ZP_05534308.1 1860 Streptomyces (NZ_ACEZ01000169.1: viridochromogenes 8822 . . . 10924) DSM 40736 256785123 NZ_ACEY01000098.1: 1660 ZP_05523554.1 1861 Streptomyces 97187 . . . 99280 lividans TK24 29829365 NC_003155.4: 1661 NP_823999.1 1862 Streptomyces 3467325 . . . 3469415 avermitilis MA-4680 260646824 FN554889.1: 1662 CBG69921.1 1863 Streptomyces 3224239 . . . 3226344 scabiei 87.22 239982381 complement 1663 ZP_04704905.1 1864 Streptomyces (NZ_ABYC01000362.1: albus 40088 . . . 42163) J1074 254382385 complement 1664 ZP_04997745.1 1865 Streptomyces (NZ_DS570390.1: sp. Mg1 111950 . . . 114034) 256769973 complement 1665 ZP_05509147.1 1866 Streptomyces (NZ_ACEW01000403.1: sp. C 16454 . . . 18541) 256776255 NZ_ACEX01000277.1: 1666 ZP_05514718.1 1867 Streptomyces 2545 . . . 4641 hygroscopicus ATCC 53653 254378850 NZ_DS570550.1: 1667 ZP_04994290.1 1868 Streptomyces 40417 . . . 42507 sp. SPB74 229854086 complement 1668 ZP_04474082.1 1869 Streptosporangium (NZ_ABUU01000066.1: roseum 39364 . . . 41415) DSM 43021 145596204 NC_009380.1: 1669 YP_001160501.1 1870 Salinispora 4234932 . . . 4237007 tropica CNB-440 159039600 NC_009953.1: 1670 YP_001538853.1 1871 Salinispora 4631907 . . . 4633976 arenicola CNS-205 238060866 NZ_GG657738.1: 1671 ZP_04605575.1 1872 Micromonospora 2330097 . . . 2332163 sp. ATCC 39149 116671783 complement 1672 YP_832716.1 1873 Arthrobacter (NC_008541.1: sp. FB24 3648900 . . . 3651011) 148807608 complement 1673 ABR13603.1 1874 Arthrobacter (EF6018801: oxydans 72 . . . 2150) 239916738 complement 1674 YP_002956296.1 1875 Micrococcus (NC_012803.1: luteus 190981 . . . 193056) NCTC 2665 255326162 NZ_ACVO01000004.1: 1675 ZP_05367249.1 1876 Rothia 127571 . . . 129661 mucilaginosa ATCC 25296 184199797 complement 1676 YP_001854004.1 1877 Kocuria (NC_010617.1: rhizophila 164688 . . . 166781) DC2201 254368446 NZ_DS264133.1: 1677 ZP_04984463.1 1878 Francisella 69966 . . . 72062 tularensis subsp. holarctica FSC022 167626922 complement 1678 YP_001677422.1 1879 Francisella (NC_010336.1: philomiragia 741506 . . . 743602) subsp. philomiragia ATCC 25017 94676965 NC_007984.1: 1679 YP_588827.1 1880 Baumannia 392877 . . . 395012 cicadellinicola str. Hc (Homalodisca coagulata) P57273 BA000003.2: 1680 NP_240007.1 1881 Buchnera 189582 . . . 191708 aphidicola str. APS (Acyrthosiphon pisum) 254444018 NZ_DS990592.1: 1681 ZP_05057494.1 1882 Verrucomicrobiae 1298899 . . . 1201010 bacterium DG1235 171914782 NZ_ABIZ01000001.1: 1682 ZP_02930252.1 1883 Verrucomicrobium 6593044 . . . 6595128 spinosum DSM4136 114777389 NZ_AATS01000006.1: 1683 ZP_01452386.1 1884 Mariprofundus 50467 . . . 52602 ferrooxydans PV-1 94500866 NZ_AAQH01000011.1: 1684 ZP_01307392.1 1885 Bermanella 33324 . . . 35456 marisrubri
(174) In other embodiments, a polynucleotide, gene and/or polypeptide encoding phosphotransacetylase can have at least about 70% to about 75%, about 80% to about 85%, about 85% to about 90%, about 90% to about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity to any one of the sequences of Tables 10 or 12a or 12b, wherein the polynucleotide, gene and/or polypeptide encodes a polypeptide having phosphotransacetylase activity.
(175) In embodiments, a polynucleotide, gene and/or polypeptide encoding phosphotransacetylase corresponds to the Enzyme Commission Number EC 2.3.1.8.
(176) In other embodiments, the phosphotransacetylase polynucleotide, gene and/or polypeptide sequences described herein or those recited in the art can be used to identify phosphotransacetylase sequences or phosphotransacetylase homologs in other cells, as described above for PDC.
(177) Methods for gene expression in recombinant host cells, including, but not limited to, yeast cells are known in the art (see, for example, Methods in Enzymology, Volume 194, Guide to Yeast Genetics and Molecular and Cell Biology (Part A, 2004, Christine Guthrie and Gerald R. Fink (Eds.), Elsevier Academic Press, San Diego, Calif.). In embodiments, the coding region for the phosphoketolase and/or phosphotransacetylase genes to be expressed can be codon optimized for the target host cell, as well known to one skilled in the art. Expression of genes in recombinant host cells, including but not limited to yeast cells, can require a promoter operably linked to a coding region of interest, and a transcriptional terminator. A number of promoters can be used in constructing expression cassettes for genes, including, but not limited to, the following constitutive promoters suitable for use in yeast: FBA1, TDH3 (GPD), ADH1, and GPM1; and the following inducible promoters suitable for use in yeast: GAL1, GAL10 and CUP1. Other yeast promoters include hybrid promoters UAS(PGK1)-FBA1p (SEQ ID NO: 1893), UAS(PGK1)-ENO2p (SEQ ID NO: 1894), UAS(FBA1)-PDC1p (SEQ ID NO: 1895), UAS(PGK1)-PDC1p (SEQ II) NO: 1896), and UAS(PGK)-OLE1p (SEQ ID NO: 1897). Suitable transcriptional terminators that can be used in a chimeric gene construct for expression include, but are not limited to, FBA1t, TDH3t, GPM1t. ERG10t, GAL1t, CYC1t, and ADH1t.
(178) Recombinant polynucleotides are typically cloned for expression using the coding sequence as part of a chimeric gene used for transformation, which includes a promoter operably linked to the coding sequence as well as a ribosome binding site and a termination control region. The coding region may be from the host cell for transformation and combined with regulatory sequences that are not native to the natural gene encoding phosphoketolase and/or phosphotransacetylase. Alternatively, the coding region may be from another host cell.
(179) Vectors useful for the transformation of a variety of host cells are common and described in the literature. Typically the vector contains a selectable marker and sequences allowing autonomous replication or chromosomal integration in the desired host. In addition, suitable vectors can comprise a promoter region which harbors transcriptional initiation controls and a transcriptional termination control region, between which a coding region DNA fragment may be inserted, to provide expression of the inserted coding region. Both control regions can be derived from genes homologous to the transformed host cell, although it is to be understood that such control regions can also be derived from genes that are not native to the specific species chosen as a production host.
(180) In embodiments, suitable promoters, transcriptional terminators, and phosphoketolase and/or phosphotransacetylase coding regions can be cloned into E. coli-yeast shuttle vectors, and transformed into yeast cells. Such vectors allow strain propagation in both E. coli and yeast strains, and can contain a selectable marker and sequences allowing autonomous replication or chromosomal integration in the desired host. Typically used plasmids in yeast include, but are not limited to, shuttle vectors pRS423, pRS424, pRS425, and pRS426 (American Type Culture Collection, Rockville, Md.), which contain an E. coli replication origin (e.g., pMB1), a yeast 2-micron origin of replication, and a marker for nutritional selection. The selection markers for these four vectors are 141S3 (vector pRS423), TRP1 (vector pRS424), LEU2 (vector pRS425) and URA3 (vector pRS426).
(181) In embodiments, construction of expression vectors with a chimeric gene encoding the described phosphoketolases and/or phosphotransacetylases can be performed by the gap repair recombination method in yeast. In embodiments, a yeast vector DNA is digested (e.g., in its multiple cloning site) to create a gap in its sequence. A number of insert DNAs of interest are generated that contain an approximately 21 by sequence at both the 5 and the 3 ends that sequentially overlap with each other, and with the 5 and 3 terminus of the vector DNA. For example, to construct a yeast expression vector for Gene X, a yeast promoter and a yeast terminator are selected for the expression cassette. The promoter and terminator are amplified from the yeast genomic DNA, and Gene X is either PCR amplified from its source organism or obtained from a cloning vector comprising Gene X sequence. There is at least a 21 by overlapping sequence between the 5 end of the linearized vector and the promoter sequence, between the promoter and Gene X, between Gene X and the terminator sequence, and between the terminator and the 3 end of the linearized vector. The gapped vector and the insert DNAs are then co-transformed into a yeast strain and plated on the medium containing the appropriate compound mixtures that allow complementation of the nutritional selection markers on the plasmids. The presence of correct insert combinations can be confirmed by PCR mapping using plasmid DNA prepared from the selected cells. The plasmid DNA isolated from yeast (usually low in concentration) can then be transformed into an E. coli strain, e.g. TOP10, followed by mini preps and restriction mapping to further verify the plasmid construct. Finally the construct can be verified by sequence analysis.
(182) Like the gap repair technique, integration into the yeast genome also takes advantage of the homologous recombination system in yeast. In embodiments, a cassette containing a coding region plus control elements (promoter and terminator) and auxotrophic marker is PCR-amplified with a high-fidelity DNA polymerase using primers that hybridize to the cassette and contain 40-70 base pairs of sequence homology to the regions 5 and 3 of the genomic area where insertion is desired. The FCR product is then transformed into yeast and plated on medium containing the appropriate compound mixtures that allow selection for the integrated auxotrophic marker. For example, to integrate Gene X into chromosomal location Y, the promoter-coding region X-terminator construct is PCR amplified from a plasmid DNA construct and joined to an, autotrophic marker (such as URA3) by either SOE PCR or by common restriction digests and cloning. The full cassette, containing the promoter-coding regionX-terminator-URA3 region, is PCR amplified with primer sequences that contain 40-70 by of homology to the legions 5 and 3 of location Y on the yeast chromosome. The PCR product is transformed into yeast and selected on growth media lacking, uracil. Transformants can be verified either by colony PCR or by direct sequencing of chromosomal DNA.
(183) The presence of phosphoketolase and phosphotransacetylase activity in the recombinant host cells disclosed herein can be, confirmed using routine methods known in the art. In a non-limiting example, and as described in the Examples herein, transformants can be screened by PCR using primers for the phosphoketolase and phosphotransacetylase genes. In embodiments, and as described in the Examples herein, transformants can be screened by PCR with primers N1039 and N1040 (SEQ ID NOs: 639 and 640) to confirm integration of the xpk1 gene, and primers N1041 and N1042 (SEQ ID NOs: 641 and 642) can be used to confirm integration of the eutD gene. In another non-limiting example, and as described in the Examples herein, transformants can be, screened for integration of phosphoketolase constructs and/or phosphotransacetylase constructs at the pdc1::ilvD(Sm) locus by the loss of ilvD(Sm) in the host cells.
(184) In another non-limiting example, and as described in the examples herein, phosphoketolase activity can be assayed by expressing phosphoketolase identifiable by the methods disclosed herein in a recombinant host cell disclosed herein that lacks endogenous phosphoketolase activity. If phosphoketolase activity is present, such cells exhibit a reduced or eliminated requirement for exogenous two-carbon substrate supplementation for growth in culture.
(185) In another non-limiting example, and as described in the examples herein, phosphoketolase and phosphotransacetylase activity can be assayed by expressing phosphoketolase and phosphotransacetylase activity identifiable by the methods disclosed herein in a recombinant host cell disclosed herein that lacks endogenous phosphoketolase and phosphotransacetylase activity. If phosphoketolase activity and phosphoketolase activity are present, such cells exhibit a reduced or eliminated requirement for exogenous two-carbon substrate supplementation for growth in culture.
(186) In another non-limiting example, phosphoketolase, and/or phosphotransacetylase activity can be confirmed by more indirect methods, such as by assaying for a downstream product in a pathway requiring phosphoketolase activity. For example, a polypeptide having phosphoketolase activity can catalyze the conversion of xylulose-5-phosphate into glyceraldehyde-3-phosphate and acetyl-phosphate and/or the conversion of fructose-6-phosphate into erythrose-4-phosphate and acetyl-phosphate. Also, a polypeptide having phosphotransacetylase activity can catalyze the conversion of acetyl-phosphate into acetyl-CoA.
(187) Suitable Pathway Carbon Substrates and Exogenous Two-Carbon Substrate Supplementation
(188) PDC-KO cells fail to grow in glucose-containing media (e.g., 2% glucose), but PDC-KO cells carrying a functional butanediol biosynthetic pathway have been shown to grow on glucose supplemented with exogenous two-carbon substrates such as ethanol (see for example, US Patent Application Publication No. 20090305363, herein incorporated by reference). In embodiments, the host cells disclosed herein can be grown in fermentation media which contains a suitable pathway carbon substrate and two-substrate supplement, including combinations of suitable pathway carbon substrates with C2-substrate supplement. Non-limiting examples of suitable pathway carbon substrates include, but are not limited to, monosaccharides such as fructose, oligosaccharides such as lactose maltose, galactose, or sucrose, polysaccharides such as starch or cellulose or mixtures thereof and unpurified mixtures from renewable feedstocks such as cheese whey permeate, cornsteep liquor, sugar beet molasses, and barley malt, including any combinations thereof. In other embodiments, the suitable pathway carbon substrates can include lactate, glycerol, or combinations thereof.
(189) In embodiments, a suitable carbon substrate can be a one-carbon substrate such as carbon dioxide, or methanol for which metabolic conversion into key biochemical intermediates has been demonstrated, or combinations thereof. In other embodiments related to methylotrophic organisms, the carbon substrate can be carbon containing compounds such as methylamine, glucosamine and a variety of amino acids for metabolic activity. In a non-limiting example, methylotrophic yeasts are known to utilize the carbon from methylamine to form trehalose or glycerol (Bellion et al., Microb. Growth C1 Compd., [Int. Symp.], 7th (1993), 415-32, Editor(s): Murrell, J. Collin; Kelly Don P. Publisher: Intercept, Andover, UK). In another non-limiting example, various species of Candida can metabolize alanine (Sulter et al., Arch. Microbiol. 153:485-489 (1990)). Hence it is contemplated that the source of carbon utilized in the present invention can encompass a wide variety of carbon containing substrates and will only be limited by the choice of organism.
(190) In other embodiments. the suitable pathway carbon substrate can be glucose, fructose. and sucrose, or mixtures of these with five-ca hon (C5) sugars such as xylose and/or arabinose for yeasts cells modified to use C5 sugars. In embodiments. sucrose can be derived from renewable sugar sources such as sugar cane, sugar beets, cassaya; sweet sorghum and mixtures thereof. In other embodiment. glucose and dextrose can derived from renewable grain sources through saccharification of starch based feedstocks including grains such as corn, wheat, rye, barley, oats. and mixtures thereof. In embodiments, the pathway carbon substrates can be derived from renewable cellulosic or lignocellulosic biomass through processes of pretreatment and saccharification, as described, for example, in U.S. Patent Application Publication No. US 20070031918 A1, which is herein incorporated by reference.
(191) As used herein, biomass refers to any cellulosic or lignocellulosic material and includes, but is not limited to, materials comprising cellulose, and optionally further comprising hemicellulose, lignin, starch, oligosaccharides and/or monosaccharides. In embodiments, biomass can also comprise additional components, such as protein and/or lipid. In other embodiments, biomass can be derived from a single source, or biomass can comprise a mixture derived from more than one source; for example, biomass can comprise a mixture of corn cobs and corn stover, or a mixture of grass and leaves. Biomass includes, but is not limited to, bioenergy crops, agricultural residues, municipal solid waste, industrial solid waste, sludge from paper manufacture, yard waste, wood and forestry waste. Other non-limiting examples of biomass include, but are not limited to, corn grain, corn cobs, crop residues such as corn husks, corn stover, grasses, wheat, wheat straw, barley, barley straw. hay, rice straw, switchgrass, waste paper, sugar cane bagasse. sorghum, soy, components obtained from milling of grains, trees, branches, roots, leaves, wood chips, sawdust, shrubs and bushes, vegetables, fruits, flowers, animal manure and mixtures thereof.
(192) The recombinant host cells described herein can be cultured using standard laboratory techniques known in the art (see, e.g. Methods in Yeast Genetics, 2005, Cold Spring Harbor Laboratory Press, Cold Spring harbor, NY, pp. 201-202). In embodiments related to media supplemented with exogenous two-carbon substrates, and as described in, the Examples, recombinant host cells can be grown in synthetic complete medium supplemented with one or more exogenous two-carbon substrates as described herein at a concentration of about 0.01%, about 0.05%, about 0.1%, about 0.5%, about 1.0%, about 1.5%, about 1.5% or about 2% (v/v) of the media. In embodiments, the recombinant host cells can be grown in synthetic complete culture without uracil or histidine, supplemented with 0.5% (v/v) ethanol. In embodiments related to growth in media that is not supplemented with exogenous two-carbon substrates, the recombinant host cells described herein can be first grown in culture medium comprising an exogenous two carbon substrate and then diluted (e.g., starting OD=0.1, ml medium in a 125 ml vented flask) into media that is not supplemented with exogenous two-carbon substrate.
(193) The growth of the recombinant, host cells described herein can be measured by methods known in the art (see, e.g., Methods in Yeast Genetics, 2005, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., pp. 201-202). In a non-limiting example, the growth of the recombinant host cells described herein can be determined by measuring the optical density (OD) of cell cultures over time. For example, the OD at 600 nm for a yeast culture is proportional to yeast cell number. In another non-limiting example, the growth of the recombinant host cells described herein can be determined by counting viable cells in a sample of the culture over time.
(194) Applicants have provided cells that have a reduced or eliminated requirement for two-carbon substrate supplementation for growth. In embodiments, such cells comprise (i) a deletion, mutation, and/or substitution in an endogenous gene encoding, a polypeptide that converts pyruvate to acetaldehyde, acetyl-phosphate or acetyl-CoA that results in a requirement for exogenous two-carbon substrate supplementation for optimal growth; (ii) a heterologous polynucleotide encoding a polypeptide having phosphoketolase activity; and optionally (iii) a heterologous polynucleotide encoding a polypeptide having phosphotransacetylase activity. In embodiments, such cells comprise (i) a modification in an endogenous polypeptide having PDC activity which results in reduced or eliminated PDC activity; (ii) a heterologous polynucleotide encoding a polypeptide having phosphoketolase activity; and optionally (iii) a heterologous polynucleotide encoding a polypeptide having phosphotransacetylase activity. As such, Applicants have also provided methods of improving the growth of a recombinant host cell comprising at least one modification in an endogenous polypeptide that converts pyruvate to acetaldehyde, acetyl-phosphate or acetyl-CoA that results in a requirement for exogenous two-carbon substrate supplementation for optimal growth comprising transforming the host cell with a heterologous polynucleotide encoding a polypeptide having phosphoketolase activity. Applicants have also provided methods of improving the growth of a recombinant host cell comprising at least one, modification in an endogenous polypeptide having pyruvate decarboxylase activity (e.g., having at least one deletion, mutation or substitution in an endogenous gene encoding a polypeptide having PDC activity that results in reduced or eliminated PDC activity) comprising transforming the host cell with a heterologous polynucleotide encoding a polypeptide having phosphoketolase activity. In other embodiments, the method further comprises transforming, a recombinant host cell described herein with a heterologous polynucleotide encoding a polypeptide having phosphotransacetylase activity.
(195) Applicants have also provided methods of reducing or eliminating the requirement for an exogenous two-carbon substrate for the growth of a recombinant host cell comprising at least one modification in an endogenous activity that converts pyruvate to acetaldehyde, acetyl-phosphate or acetyl-CoA that results in a requirement for exogenous two-carbon substrate supplementation for optimal growth comprising transforming the host cell with a heterologous polynucleotide encoding a polypeptide having phosphoketolase activity comprising, transforming the recombinant host cell with a heterologous polynucleotide encoding a polypeptide having phosphoketolase activity. In other embodiments, the method further comprises transforming the recombinant host cell with a heterologous polynucleotide encoding a polypeptide having phosphotransacetylase activity.
(196) Applicants have also provided methods of reducing the requirement for an exogenous two-carbon substrate for the growth of a recombinant host cell comprising at least one modification in an endogenous polypeptide having. PDC activity (e.g., having at least one deletion, mutation or substitution in an endogenous gene encoding a polypeptide having pyruvate decarboxylase activity) comprising transforming the recombinant host cell with a heterologous polynucleotide encoding a polypeptide having phosphoketolase activity. In other embodiments, the method further comprises transforming the recombinant host cell with a heterologous polynucleotide encoding a polypeptide having phosphotransacetylase activity.
(197) In addition, Applicants have provided methods of eliminating the requirement for an exogenous two-carbon substrate for the growth of a recombinant host cell comprising at least one modification in an endogenous polypeptide having PDC activity (e.g., having at least one deletion, mutation or substitution in an endogenous gene encoding a polypeptide having PDC activity that results in reduced or eliminated PDC activity) comprising transforming the recombinant host cell with a heterologous polynucleotide encoding a polypeptide having phosphoketolase activity. In other embodiments, the method further comprises transforming the recombinant host cell with a heterologous polynucleotide encoding a polypeptide having phosphotransacetylase activity.
(198) In embodiments, a reduced requirement for exogenous two-carbon substrate supplementation can be a growth rate of the recombinant host cells described herein in media that is not supplemented, with an exogenous two-carbon substrate that is the same or substantially equivalent to the growth rate of a recombinant host cell comprising a modification in an endogenous activity that converts pyruvate to acetaldehyde, acetyl-phosphate or acetyl-CoA grown in media that is supplemented with an exogenous two-carbon substrate. In embodiments, such a growth rate can be at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% of the growth rate of a recombinant host cell comprising a modification in an endogenous activity that converts pyruvate to acetaldehyde, acetyl-phosphate or acetyl-CoA grown in media that is supplemented with an exogenous two-carbon substrate.
(199) In embodiments, a reduced requirement for exogenous two-carbon substrate supplementation can be a growth rate of the recombinant host cells described herein in media that is not supplemented with an exogenous two-carbon substrate that is the same or substantially equivalent to the growth rate of a recombinant host cell comprising a modification in an endogenous PDC activity grown in media that is supplemented with an exogenous two-carbon substrate. In embodiments, such a growth rate can be at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% of the growth rate of a recombinant host cell comprising a modification in an endogenous PDC activity grown in media that is supplemented with an exogenous two-carbon substrate.
(200) In other embodiments, the recombinant host cells described herein have a growth rate in media that is not supplemented with an exogenous two-carbon substrate that is greater than the growth rate of a recombinant host cell comprising a modification in an endogenous activity that converts pyruvate to acetaldehyde, acetyl-phosphate or acetyl-CoA in media that is not supplemented with an exogenous two-carbon substrate.
(201) In other embodiments, the recombinant host cells described herein have a growth rate in media that is not supplemented with an exogenous two-carbon substrate that is greater than the growth rate of a recombinant host cell comprising a modification in an endogenous PDC activity in media that is not supplemented with an exogenous two-carbon substrate.
(202) In other embodiments, the recombinant host cells described herein can have an increased glucose consumption compared to a recombinant host cell comprising a modification in an endogenous polypeptide that converts pyruvate to acetaldehyde, acetyl-phosphate or acetyl-CoA.
(203) In other embodiments, the recombinant host cells described herein can have an increased glucose consumption compared to a recombinant host cell comprising a modification in an endogenous polypeptide having PDC activity (e.g., at least one deletion, mutation, and/or substitution in an endogenous gene encoding a polypeptide having PDC activity that reduces or eliminates PDC activity).
(204) Glucose consumption of the recombinant host cells described herein can be measured by methods known in the art (see, e.g., Methods in Yeast Genetics, 2005, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., pp. 201-202). In a non-limiting example, glucose consumption can be measured by quantitating the amount of glucose in culture media by HPLC or with a YSI Biochemistry Analyzer (YSI, Inc., Yellow Springs, Ohio).
(205) In other embodiments, methods of producing a recombinant host cell are provided comprising transforming a recombinant host cell comprising a modification in an endogenous polynucleotide, gene or polypeptide encoding pyruvate decarboxylase (e.g., at least one deletion, mutation, and/or substitution in an endogenous gene encoding a polypeptide having pyruvate decarboxylase activity) with a heterologous polynucleotide encoding a polypeptide having phosphoketolase activity. In other embodiments, the method further comprises transforming the recombinant host cell with a heterologous polynucleotide encoding a polypeptide having phosphotransacetylase activity.
(206) In other embodiments, methods for the conversion of xylulose 5-phosphate or fructose 6-phosphate into acetyl-phosphate are provided comprising (i) providing a recombinant host cell as described herein, or combinations thereof; and (ii) growing the recombinant host cell under conditions wherein xylulose 5-phosphate or fructose-6-phosphate is converted into acetyl-phosphate. In other embodiments, methods for the conversion of xylulose 5-phosphate or fructose-6-phosphate into acetyl-CoA are provided comprising (i) providing a recombinant rose cell as described herein, or combinations thereof; and (ii) growing the recombinant host cell under conditions where xylulose 5-phosphate or fructose-6-phosphate is converted into acetyl-CoA.
(207) In other embodiments, methods for the conversion of acetyl-phosphate to, acetyl-CoA are provided comprising (i) providing a recombinant host cell as described herein, or combinations thereof; and (ii) growing the recombinant host cell under conditions where acetyl-phosphate is converted into acetyl-CoA. In other embodiments, methods for increasing the specific activity of a heterologous polypeptide having phosphoketolase activity in a recombinant host cell are provided comprising (i) providing a recombinant host cell as described herein, or combinations thereof; and (ii) growing the recombinant host cell under conditions wherein the heterologous polypeptide having phosphoketolase activity is expressed in functional form having a specific activity greater than the same recombinant host cell lacking the heterologous polypeptide having phosphoketolase activity.
(208) In other embodiments, methods for increasing the specific activity of a heterologous polypeptide having phosphotransacetylase activity in a recombinant host cell are provided comprising (i) providing a recombinant host cell described herein, or combinations thereof; and (ii) growing the recombinant host cell under conditions whereby the heterologous polypeptide having phosphotransacetylase activity is expressed in functional form having a specific activity greater than the same recombinant host cell lacking a heterologous polypeptide having phosphotransacetylase activity.
(209) In still other embodiments, methods for increasing the activity of the phosphoketolase pathway in a recombinant host cell are provided comprising (i) providing a recombinant host cell as described herein, or combinations thereof; and (ii) growing the host cell under conditions whereby the activity of the phosphoketolase pathway in the host cell is increased.
(210) Threonine aldolase (E.C. number 4.1.2.5) catalyzes cleavage of threonine to produce glycine and acetaldehyde. Plasmid-based overexpression of a gene encoding this enzyme in S. cerevisiae PDC-KO strains was shown to eliminate the requirement for exogenous C2 supplementation (van Maris et al, Appl Environ Microbiol. 2003 April; 69(4):2094-9). In embodiments, recombinant host cells comprise (i) a deletion, mutation, and/or substitution in an endogenous gene encoding a polypeptide that converts pyruvate to acetaldehyde, acetyl-phosphate or acetyl-CoA that results in a requirement for exogenous two-carbon substrate supplementation for optimal growth; and (ii) heterologous polynucleotide encoding a polypeptide having threonine aldolase activity.
(211) Engineered Biosynthetic Pathways Using Pyruvate
(212) In embodiments, the recombinant host cells described herein can be engineered to have a biosynthetic pathway for production of a product from pyruvate. A product from such a pyruvate-utilizing biosynthetic pathway includes, but is not limited to, 2,3-butanediol, isobutanol, 2-butanol, 2-butanone, valine, leucine, alanine, lactic acid, malic acid, fumaric acid, succinic acid and isoamyl alcohol. The features of any pyruvate-utilizing biosynthetic pathway may be engineered in the recombinant host cells described herein in any order. Any product made using a biosynthetic pathway that has, pyruvate as the initial substrate can be produced with greater effectiveness in a recombinant host cell disclosed herein having a, modification in an endogenous polypeptide that converts pyruvate to acetaldehyde, acetyl-phosphate or acetyl-CoA (such as pyruvate decarboxylase, pyruvate formate lyase, pyruvate dehydrogenase, pyruvate oxidase, or pyruvate:ferredoxin oxioreductase) and having heterologous phosphoketolase and/or phosphotransacetylase activity, compared to a recombinant host cell having a modification in an endogenous polypeptide that converts pyruvate to acetaldehyde, acetyl-phosphate or acetyl-CoA (such as pyruvate decarboxylase, pyruvate formate lyase, pyruvate dehydrogenase, pyruvate oxidase, or pyruvate:ferredoxin oxioreductase). Any product made using a biosynthetic pathway that has pyruvate as the initial substrate can be produced with greater effectiveness in a recombinant host cell disclosed herein having a modification in an endogenous polypeptide having PDC activity that reduces or eliminates PDC activity and having heterologous phosphoketolase and/or phosphotransacetylase activity, compared to a recombinant host cell having a modification in an endogenous polypeptide having PDC activity that reduces or eliminates PDC activity.
(213) The biosynthetic pathway of the recombinant host cells described herein can be any pathway that utilizes pyruvate and produces a desired product. The pathway genes may include endogenous genes and/or heterologous genes. Typically at least one gene in the biosynthetic pathway is a heterologous gene. Suitable biosynthetic pathways for production of butanol are known in the art, and certain suitable pathways are described herein. In some embodiments, the butanol biosynthetic pathway comprises, at least one gene that is heterologous to the host cell. In some embodiments, the butanol biosynthetic pathway comprises more than one gene that is heterologous to the host cell. In some embodiments, the butanol biosynthetic pathway comprises heterologous genes encoding polypeptides corresponding to every step of a biosynthetic pathway.
(214) Genes and polypeptides that can be used for substrate to product conversions described herein as well as methods of identifying such, genes and polypeptides, are described herein and/or in the art, for example, for isobutanol, in the Examples and in U.S. Pat. No. 7,851,188. Ketol-acid reductoisomerase (KARI) enzymes are described in U.S. Patent Appl. Pub. Nos. 20080261230 A1, 20090163376 A1, 20100197519 A1, and PCT Appl. Pub. No. WO/2011/04.1415. Examples of KARIs disclosed therein are those from Lactococcus lactis, Vibrio cholera, Pseudomonas aeruginosa PAO1, and Pseudomonas fluorescens PF5 mutants. KARIs include Anaerostipes caccae KARI variants K9G9 and K9D3 (SEQ ID NOs: 1911 and 1910, respectively). US Appl. Pub. No. 20100081154 A1, and U.S. Pat. No. 7,851,188 describe dihydroxyacid dehydratases (DHADs), including a DHAD from Streptococcus mutans. U.S. Patent Appl. Publ. No. 20090269823 A1 describes SadB, an alcohol dehydrogenase (ADH) from Achromobacter xylosoxidans. Alcohol dehydrogenases also include horse liver ADH and Beijerinkia indica ADH (protein SEQ ID NO: 1923).
(215) An example of a biosynthetic pathway for producing 2,3-butanediol can be engineered in the recombinant host cells described herein, as described in U.S. Patent Application No. 20090305363, which is herein incorporated by reference. The 2,3-butanediol pathway is a portion of the 2-butanol biosynthetic pathway that is disclosed in U.S. Patent Application Publication No. US 20070292927 A1, which is herein incorporated by reference. Such pathway steps include, but are not limited to, conversion of pyruvate to acetolactate by acetolactate synthase, conversion of acetolactate to acetoin by acetolactate decarboxylase, and conversion of acetoin to 2,3-butanediol by butanediol dehydrogenase. The skilled person will appreciate that polypeptides having the activity of such pathway steps can be isolated from a variety of sources can be used in the recombinant host cells described herein.
(216) In addition, examples of biosynthetic pathways for production of 2-butanone or 2-butanol that can be engineered in the recombinant host cells described herein are disclosed in U.S. Patent Application Publication Nos. US 20070292927 A1 and US 20070259410 A1, which are herein incorporated by reference. The pathway in U.S. Patent Application Publication No. US 20070292927 A1 is the same as described for butanediol production with the addition of the following steps: 2,3-butanediol to 2-butanone as catalyzed for example by diol dehydratase or glycerol dehydratase; and 2-butanone to 2-butanol as catalyzed for example by butanol dehydrogenase.
(217) Described in U.S. Patent Application Publication No. US 20090155870 A1, which is herein incorporated by reference, is the construction of chimeric genes and genetic engineering of yeast for 2-butanol production using the U.S. Patent Application Publication No. US 20070292927 A1 disclosed biosynthetic pathway. Further description for gene construction and expression related to these pathways can be found, for example, in International Publication No. WO 2009046370 (e.g., butanediol dehydratases); and U.S. Patent Application Publication No. US 20090269823 A1 (e.g., butanol dehydrogenase) and U.S. Patent Application Publication No. US 20070259410 A1 which are herein incorporated by reference. The skilled person will appreciate that polypeptides having the activity of such pathway steps can be isolated from a variety of sources can be used in the recombinant host cells described herein.
(218) Biosynthetic pathways for the production of isobutanol that may be used include those described in U.S. Pat. No. 7,851,188 and PCT Publication WO 2007050671, incorporated herein by reference. One isobutanol biosynthetic pathway comprises the following substrate to product conversions: pyruvate to acetolactate, which may be catalyzed, for example, by acetolactate synthase; acetolactate to 2,3-dihydroxyisovalerate, which may be catalyzed, for example, by acetohydroxy acid reductoisomerase; 2,3-dihydroxyisovalerate to -ketoisovalerate, which may be catalyzed, for example, by acetohydroxy acid dehydratase; -ketoisovalerate to isobutyraldehyde, which may be catalyzed, for example, by a branched-chain keto acid decarboxylase; and isobutyraldehyde to isobutanol, which may be catalyzed, for example, by a branched-chain alcohol dehydrogenase. In some embodiments, the isobutanol biosynthetic pathway comprises at least one gene, at least two genes, at least three genes, or at least four genes that is/are heterologous to the yeast cell. In embodiments, each substrate to product conversion of an isobutanol biosynthetic pathway in a recombinant host cell is catalyzed by a heterologous polypeptide. In embodiments, the polypeptide catalyzing the substrate to product conversions of acetolactate to 2,3-dihydroxyisovalerate and/or the polypeptide catalyzing the substrate to product conversion of isobutyraldehyde to isobutanol are capable of utilizing NADH as a cofactor.
(219) An example of a biosynthetic pathway for production of valine that can be engineered in the recombinant host cells described herein includes the steps of acetolactate conversion to 2,3-dihydroxy-isovalerate by acetohydroxyacid reductoisomerase (ILV5), conversion of 2,3-dihydroxy-isovalerate to 2-keto-isovalerate by dihydroxy-acid dehydratase (ILV3), and conversion of 2-keto-isovalerate to valine by branched-chain, amino acid transaminase (BAT2) and branched-chain amino acid aminotransferase (BAT1). Biosynthesis of leucine includes the same steps to 2-keto-isovalerate, followed by conversion of 2-keto-isovalerate to alpha-isopropylmalate by alpha-isopropylmalate synthase (LEU9. LEU4), conversion of alpha-isopropylmalate to beta-isopropylmalate by isopropylmalate isomerase (LEU1), conversion of beta-isopropylmalate to alpha-ketoisocaproate by beta-IPM dehydrogenase (LEU2), and finally conversion of alpha-ketoisocaproate to leucine by branched-chain amino acid transaminase (BAT2) and branched-chain amino acid aminotransferase (BAT1). It is desired for production of valine or leucine to overexpress at least one of the enzymes in these described pathways.
(220) An example of a biosynthetic pathway for production of isoamyl alcohol that can be engineered in the recombinant host cells described herein includes the steps of leucine conversion to alpha-ketoisocaproate by branched-chain amino acid transaminase (BAT2) and branched-chain amino acid aminotransferase (BAT1), conversion of alpha-ketoisocaproate to 3-methylbutanal by ketoisocaproate decarboxylase (THI3) or decarboxylase ARO10, and finally conversion of 3-methylbutanal to isoamyl alcohol by an alcohol dehydrogenase such as ADH1 or SFA1. Production of isoamyl alcohol benefits from increased production of leucine or the alpha-ketoisocaproate intermediate by overexpression of one or more enzymes in biosynthetic pathways for these chemicals. In addition, one or both enzymes for the final two steps can be overexpressed.
(221) An example of a biosynthetic pathway for production of lactic acid that can be engineered in the recombinant host cells described herein includes pyruvate conversion to lactic acid by lactate dehydrogenase. Engineering yeast for lactic acid production using lactate dehydrogenase, known as EC 1.1.1.27, is well known in the art such as in Ishida et al. (Appl. Environ. Microbiol. 71:1964-70 (2005)).
(222) An example of a biosynthetic pathway for production of alanine that can be engineered in the recombinant host cells described herein includes pyruvate conversion to alanine by aminotransferase.
(223) An example of a biosynthetic pathway for production of malate that can be engineered in the recombinant host cells described herein includes pyruvate conversion to oxaloacetate by pyruvate carboxylase, and conversion of oxaloacetate to malate by malate dehydrogenase as described in Zelle et al. (Applied and Environmental Microbiology 74:2766-77 (2008)). In addition, a malate transporter can be expressed.
(224) An example of a biosynthetic pathway for production of fumarate that can be engineered in the recombinant host cells described herein includes pyruvate conversion to oxaloacetate by pyruvate carboxylase, and conversion of oxaloacetate to malate by malate dehydrogenase as described in Zelle et al. (Applied and Environmental Microbiology 74:2766-77 (2008)). In addition, a fumarase and a fumarate transporter can be expressed. Favorable production conditions and engineering of fungi for fumarate production is well known in the art, described e.g. by Goldberg et al. (Journal of Chemical Technology and Biotechnology 81:1601-1611 (2006)).
(225) An example of a biosynthetic pathway for production of succinate that can be engineered in the recombinant host cells described herein includes pyruvate conversion to oxaloacetate by pyruvate carboxylase, and conversion of oxaloacetate to malate by malate dehydrogenase as described in Zelle et al. (Applied and Environmental Microbiology 74:2766-77 (2008)). In addition, a fumarase, a succinate dehydrogenase and a succinate transporter can be expressed.
(226) The skilled person will appreciate that polypeptides having activities of the above-mentioned biosynthetic pathways can be isolated from a variety of sources can be used in the recombinant host cells described herein.
(227) It will be appreciated that host cells comprising a butanol biosynthetic pathway such as an isobutanol biosynthetic pathway as provided herein may further comprise one or more additional modifications. U.S. Appl. Pub. No. 20090305363 (incorporated by reference) discloses increased conversion of pyruvate to acetolactate by engineering yeast for expression of a cytosol-localized acetolactate synthase and substantial elimination of pyruvate decarboxylase activity. Modifications to reduce glycerol-3-phosphate dehydrogenase activity and/or disruption in at least one gene encoding a polypeptide having pyruvate decarboxylase activity or a disruption in at least one gene encoding a regulatory element controlling pyruvate decarboxylase gene expression as described in U.S. Patent Appl. Pub. No. 20090305363 (incorporated herein by reference), modifications to a host cell that provide for increased carbon flux through an Entner-Doudoroff Pathway or reducing equivalents balance as described in U.S. Patent Appl. Pub. No. 20100120105 (incorporated herein by reference). Other modifications include integration of at least one polynucleotide encoding a polypeptide that catalyzes a step in a pyruvate-utilizing biosynthetic pathway. Other modifications include at least one deletion, mutation, and/or substitution in an endogenous polynucleotide encoding a polypeptide having acetolactate reductase activity. In embodiments, the polypeptide having acetolactate reductase activity is YMR226C (SEQ ID NO: 1912) of Saccharomyces cerevisae or a homolog thereof. Additional modifications include a deletion, mutation, and/or substitution in an endogenous polynucleotide encoding a polypeptide having aldehyde dehydrogenase and/or aldehyde oxidase activity. In embodiments, the polypeptide having aldehyde dehydrogenase activity is ALD6 (SEQ ID NO: 1909) from Saccharomyces cerevisiae or a homolog thereof. A genetic modification which has the effect of reducing glucose repression wherein the yeast production host cell is pdcis described in U.S. Appl. Publication No. 20110124060, incorporated herein by reference.
(228) Recombinant host cells may further comprise (a) at least one heterologous polynucleotide encoding a polypeptide having dihydroxy-acid dehydratase activity; and (b)(i) at least one deletion, mutation, and/or substitution in an endogenous gene encoding a polypeptide affecting Fe-S cluster biosynthesis; and/or (ii) at least one heterologous polynucleotide encoding a polypeptide affecting Fe-S cluster biosynthesis. In embodiments, the polypeptide affecting Fe-S cluster biosynthesis is encoded by AFT1 (nucleic acid SEQ ID NO: 1913, amino acid SEQ ID NO: 1914), AFT2 (SEQ ID NOs: 1915 and 1916), FRA2 (SEQ ID NOs: 1917 and 1918), GRx3(SEQ ID NOs: 1919 and 1920), or CCC1 (SEQ ID NOs: 1921 and 1922). In embodiments, the polypeptide affecting Fe-S cluster biosynthesis is constitutive mutant AFT1 L99A, AFT1 L102A, AFT1 C291F, or AFT1 C293F.
(229) Fermentation Media
(230) The recombinant host cells disclosed herein can be grown in fermentation media for production of a product utilizing pyruvate. For maximal production of some products, such as 2,3-butanediol, isobutanol, 2-butanone, or 2-butanol, the recombinant host cells disclosed herein used as production hosts preferably have enhanced tolerance to the produced chemical, and have a high rate of carbohydrate utilization. These characteristics can be conferred by mutagenesis and selection, genetic engineering, or can be natural.
(231) Fermentation media for production of the products disclosed herein may contain glucose. Additional carbon substrates for product production pathways can include but are not limited to those described above. It is contemplated that the source of carbon utilized can encompass a wide variety of carbon containing substrates and will only be limited by the choice of organism.
(232) In addition to an appropriate carbon source, fermentation media can contain suitable minerals, salts, cofactors, buffers and other components, known to those skilled in the art, suitable for the growth of the cultures and promotion of the enzymatic pathway necessary for production of the desired product.
(233) Culture Conditions
(234) 10192 Typically cells are grown at a temperature in the range of about 20 C. to about 37 C. in, an appropriate medium. Suitable growth media for the recombinant host cells described herein are common commercially prepared media such as broth that includes yeast nitrogen base, ammonium sulfate, and dextrose as the carbon/energy source) or YPD Medium, a blend of peptone, yeast extract, and dextrose in optimal proportions for growing most Saccharomyces cerevisiae strains. Other defined or synthetic growth media may also be used and the appropriate medium for growth of the particular microorganism will be known by one skilled in the art of microbiology or fermentation science.
(235) Suitable pH ranges for the fermentation are between pH 3.0 to pH 7.5, where pH 4.5 to pH 6.5 is preferred as the initial condition.
(236) Fermentations can be performed under aerobic or anaerobic conditions, where anaerobic or microaerobic conditions are preferred.
(237) The amount of product in the fermentation medium can be determined using a number of methods known in the art, for example, high performance liquid chromatography (HPLC) or gas chromatography (GC).
(238) Industrial Batch and Continuous Fermentations
(239) A batch method of fermentation can be used with the recombinant host cells described herein. A classical batch fermentation is a closed system where the composition of the medium is set at the beginning of the fermentation and not subject to artificial alterations during the fermentation. Thus, at the beginning of the fermentation the medium is inoculated with the desired organism or organisms, and fermentation is permitted to occur without adding anything to the system. Typically, however, a batch fermentation is batch with respect to the addition of carbon source and attempts are often made at controlling factors such as pH and oxygen concentration. In batch systems the metabolite and biomass compositions of the system, change constantly up to the time the fermentation is stopped. Within batch cultures cells progress through a static lag phase to a high growth log phase and finally to a stationary phase where growth rate is diminished or halted. If untreated, cells in the stationary phase will eventually die. Cells in log phase generally are responsible for the bulk of production of end product or intermediate.
(240) A Fed-Batch system can also be used with the recombinant host cells described herein. A Fed-Batch system is similar to a typical batch system with the exception that the carbon source substrate is added in increments as the fermentation progresses. Fed-Batch systems are useful when catabolite repression (e.g. glucose repression) is apt to inhibit the metabolism of the cells and where it is desirable to have limited amounts of substrate in the media. Measurement of the actual substrate concentration in Fed-Batch systems is difficult and is therefore estimated on the basis of the changes of measurable factors such as pH, dissolved oxygen and the partial pressure of waste gases such as CO.sub.2. Batch and Fed-Batch fermentations are common and well known in the art and examples may be found in Thomas D. Brock in Biotechnology: A Textbook of Industrial Microbiology, Second Edition (1989) Sinauer Associates, Inc., Sunderland, Mass., or Deshpande, Mukund V., Appl. Biochem. Biotechnol., 36:227, (1992), herein incorporated by reference.
(241) Although a batch mode can be performed, it is also contemplated that continuous fermentation methods could also be performed with the recombinant host cells described herein. Continuous fermentation is an open system where a defined fermentation medium is added continuously to a bioreactor and an equal amount of conditioned media is removed simultaneously for processing. Continuous fermentation generally maintains the cultures at a constant high density where cells are primarily in log phase growth.
(242) Continuous fermentation allows for the modulation of one factor or any number of factors that affect cell growth or end product concentration. For example, one method will maintain a limiting nutrient such as the carbon source or nitrogen level at a fixed rate and allow all other parameters to vary. In other systems a number of factors affecting growth can be altered continuously while the cell concentration, measured by media turbidity, is kept constant. Continuous systems strive to maintain steady state growth, conditions and thus the cell loss due to the medium being drawn off must be balanced against the cell growth rate in the fermentation. Methods of modulating nutrients and growth factors for continuous fermentation processes as well as techniques for maximizing the rate of product formation are well known in the art of industrial microbiology and a variety of methods are detailed by Brock, supra.
(243) It is contemplated that the present invention can be practiced using either batch, fed-batch or continuous processes and that any known mode of fermentation would be suitable. Additionally, it is contemplated that cells can be immobilized on a substrate as whole cell catalysts and subjected to fermentation conditions for production.
(244) Product Isolation from Fermentation Medium
(245) Products can be isolated from the fermentation medium by methods known to one skilled in the art. For example, bioproduced isobutanol may be isolated from the fermentation medium using methods known in the art for ABE fermentations (see, e.g., Dune, Appl. Microbiol. Biotechnol. 49:639-648 (1998), Groot et al., Process. Biochem. 27:61-75 (1992), and references therein). For example, solids may be removed from the fermentation medium by centrifugation, filtration, decantation, or the like. Then, the isobutanol may be isolated from the fermentation medium using methods such as distillation, azeotropic distillation, liquid-liquid extraction, adsorption, gas stripping, membrane evaporation, pervaporation or vacuum flash fermentation (see e.g., U.S. Pub. No. 20090171129 A1, and International Pub. No. WO2010/151832 A1, both incorporated herein by reference in their entirety).
(246) Because butanol forms a low boiling point, azeotropic mixture with, water, distillation can be used to separate the mixture up to its azeotropic composition. Distillation may be used in combination with another separation method to obtain separation around the azeotrope. Methods that may be used in combination with distillation to isolate and purify butanol include, but are not limited to, decantation, liquid-liquid extraction, adsorption, and membrane-based techniques. Additionally, butanol may be isolated using azeotropic distillation using an entrainer (see, e.g., Doherty and Malone, Conceptual Design of Distillation Systems, McGraw Hill, N.Y., 2001).
(247) The butanol-water mixture forms a heterogeneous azeotrope so that distillation may be used in combination with decantation to isolate and purify the isobutanol. In this method, the isobutanol containing fermentation broth is distilled to near the azeotropic composition. Then, the azeotropic mixture is condensed, and the isobutanol is separated from the fermentation medium by decantation. The decanted aqueous phase may be returned to the first distillation column as reflux. The isobutanol-rich decanted organic phase may be further purified by distillation in a second distillation column.
(248) The butanol can also be isolated from the fermentation medium using liquid-liquid extraction in combination with distillation. In this method, the isobutanol is extracted from the fermentation broth using liquid-liquid extraction with a suitable solvent. The isobutanol-containing organic phase is then distilled to separate the butanol from the solvent.
(249) Distillation in combination with adsorption can also be used to isolate isobutanol from the fermentation medium. In this method, the fermentation broth containing the isobutanol is distilled to near the azeotropic composition and then the remaining water is removed by use of an adsorbent. such as molecular sieves (Aden et al., Lignocellulosic Biomass to Ethanol Process Design and Economics Utilizing Co-Current Dilute Acid Prehydrolysis and Enzymatic Hydrolysis for Corn Stover, Report NREL/TP-510-32438, National Renewable Energy Laboratory, June 2002).
(250) Additionally, distillation in combination with pervaporation may be used to isolate and purify the isobutanol from the fermentation medium. In this method, the fermentation broth containing the isobutanol is distilled to near the azeotropic composition, and then the remaining water is removed by pervaporation through a hydrophilic membrane (Guo et al., J. Membr. Sci. 245, 199-210 (2004)).
(251) In situ product removal (ISPR) (also referred to as extractive fermentation) can be used to remove butanol (or other fermentative alcohol) from the fermentation vessel as it is produced, thereby allowing the microorganism to produce butanol at high yields. One method for ISPR for removing fermentative alcohol that has been described in the art is liquid-liquid extraction. In, general, with regard to butanol fermentation, for example, the fermentation medium, which includes the microorganism, is contacted with an organic extractant at a time before the butanol concentration reaches a toxic level. The organic extractant and, the fermentation medium form a biphasic mixture. The butanol partitions into the organic extractant phase, decreasing the concentration in the aqueous phase containing the microorganism, thereby limiting the exposure of the microorganism to the inhibitory butanol.
(252) Liquid-liquid extraction can be performed, for example, according to the processes described in U.S. Patent Appl. Pub. No. 20090305370, the disclosure of which is hereby incorporated in its entirety. U.S. Patent Appl. Pub. No. 20090305370 describes methods for producing and recovering butanol from a fermentation broth using liquid-liquid extraction, the methods comprising the step of contacting the fermentation broth with a water immiscible extractant to form a two-phase mixture comprising an aqueous phase and an organic phase. Typically, the extractant can be an organic extractant selected from the group consisting of saturated, mono-unsaturated, poly-unsaturated (and mixtures thereof) C.sub.12 to C.sub.22 fatty alcohols, C.sub.12 to C.sub.22 fatty acids, esters of C.sub.12 to C.sub.22 fatty acids, C.sub.12 to C.sub.22 fatty aldehydes, and mixtures thereof. The extractant(s) for ISPR can be non-alcohol extractants. The ISPR extractant can be an exogenous organic extractant such as oleyl alcohol, behenyl alcohol, cetyl alcohol, lauryl alcohol, myristyl alcohol, stearyl alcohol, 1-undecanol, oleic acid, lauric acid, myristic acid, stearic acid, methyl myristate, methyl oleate, undecanal, lauric aldehyde, 20-methylundecanal, and mixtures thereof.
(253) In some embodiments, the alcohol can be esterfied by contacting the alcohol in a fermentation medium with an organic acid (e.g., fatty acids) and a catalyst (e.g. enzyme such as a lipase) capable of esterifying the alcohol with the organic acid. In such embodiments, the organic acid can serve as an ISPR extractant into which the alcohol esters partition. The organic acid can be supplied to the fermentation vessel and/or derived from the biomass supplying fermentable carbon fed to the fermentation vessel. Lipids present in the feedstock can be catalytically hydrolyzed to organic acid, and the same catalyst (e.g., enzymes) can esterify the organic acid with the alcohol. The catalyst can be supplied to the feedstock prior to fermentation, or can be supplied to the fermentation vessel before or contemporaneously with the supplying of the feedstock. When the catalyst is supplied to the fermentation vessel, alcohol esters can be obtained by hydrolysis of the lipids into organic acid and substantially simultaneous esterification of the organic acid with butanol present in the fermentation vessel. Organic acid and/or native oil not derived from the feedstock can also be fed to the fermentation vessel, with the native oil being hydrolyzed into organic acid. Any organic acid not esterified with the alcohol can serve as part of the ISPR extractant. The extractant containing alcohol esters can be separated from the fermentation medium, and the alcohol can be recovered from the extractant. The extractant can be recycled to the fermentation vessel. Thus, in the case of butanol production, for example, the conversion of the butanol to an ester reduces the free butanol concentration in the fermentation medium, shielding the microorganism from the toxic effect of increasing butanol concentration. In addition, unfractionated grain can be used as feedstock without separation of lipids therein, since the lipids can be catalytically hydrolyzed to organic acid, thereby decreasing the rate of build-up of lipids in the ISPR extractant.
(254) In situ product removal can be carried out in a batch mode or a continuous mode. In a continuous mode of in situ product removal, product is continually removed from the reactor. In a batchwise mode of in situ product removal, a volume of organic extractant is added to the fermentation vessel and the extractant is not removed during the process. For in situ product removal, the organic extractant can contact the fermentation medium at the start of the fermentation forming a biphasic fermentation medium. Alternatively, the organic extractant can contact the fermentation medium after the microorganism has achieved a desired amount of growth, which can be determined by measuring the optical density of the culture. Further, the organic extractant can contact the fermentation medium at a time at which the product alcohol level in the fermentation medium reaches a preselected level. In the case of butanol production according to some embodiments of the present invention, the organic acid extractant can contact the fermentation medium at a time before the butanol concentration reaches a toxic level, so as to esterify the butanol with the organic acid to produce butanol esters and consequently reduce the concentration of butanol in the fermentation vessel. The ester-containing organic phase can then be removed from the fermentation vessel (and separated from the fermentation broth which constitutes the aqueous phase) after a desired effective titer of the butanol esters is achieved. In some embodiments, the ester-containing organic phase is separated from the aqueous phase after fermentation of the available fermentable sugar in the fermentation vessel is substantially complete.
EXAMPLES
(255) The meaning of abbreviations used is as follows: min means minute(s), h means hour(s), sec means second(s), l means microliter(s), ml means milliliter(s), L means liter(s), nm means nanometer(s), mm means millimeter(s), cm means centimeter(s), m means micrometer(s), mM means millimolar, M means molar, mmol means millimole(s), mole means micromole(s), g means gram(s), g means microgram(s), mg means milligram(s), rpm means revolutions per minute, w/v means weight/volume, v/v means volume/volume, OD means optical density, bp means base pair(s), and PCR means polymerase chain reaction.
(256) General Methods:
(257) Standard recombinant DNA and molecular cloning techniques used in the Examples are well known in the art and are described by Sambrook, J., Fritsch, E. F. and Mamatis, T., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989, by T. J. Sillavy, M. L. Bennan, and L. W. Enquist, Experiments' with Gene Fusions, Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y., 1984, and by Ausubel, F. M. et al., Current Protocols in Molecular Biology, Greene Publishing Assoc. and Wiley-Interscience, N.Y., 1987. Phusion HF Master Mix (NEB Cat. No. F-531) and HotStarTaq Master Mix (Qiagen Cat. No. 203443) were used for PCR in gene cloning and clone screening, respectively.
(258) Materials and methods suitable for the maintenance and growth of bacterial cultures are also well known in the art. Techniques suitable for use in the following Examples may be found in Manual of Methods for General Bacteriology, Phillipp Gerhardt, R. G. E. Murray, Ralph N. Costilow, Eugene W. Nester, Willis A. Wood, Noel R. Krieg and G. Briggs Phillips, eds., American Society for Microbiology, Washington, D.C., 1994, or by Thomas D. Brock in Biotechnology: A Textbook of Industrial Microbiology, Second Edition, Sinauer Associates, Inc., Sunderland, Mass., 1989. All reagents, restriction enzymes and materials used for the growth and maintenance of bacterial cells were obtained from Aldrich Chemicals (Milwaukee, Wis.), BD Diagnostic Systems (Sparks, Md.)), Life Technologies (Rockville, Md.), or Sigma Chemical Company (St. Louis, Mo.), unless otherwise specified.
(259) HPLC
(260) Analysis for fermentation by-product composition is well known to those skilled in the art. For example, one high performance liquid chromatography (HPLC) method utilizes a Shodex SH-1011 column with a Shodex SH-G guard column (both available from Waters Corporation, Milford, Mass.), with refractive index (RI) detection. Chromatographic separation is achieved using 0.01 M H.sub.2SO.sub.4 as the mobile phase with a flow rate of 0.5 mL/min and a column temperature of 50 C. Isobutanol retention time is 47.6 minutes. For butanediol, meso-butanediol eluted at 26.0 min and 2R,3R-butanediol eluted at 27.7 min.
Example 1
Construction of phosphoketolase/phosphotransacetylase expression cassette
(261) The xpk1 and eutD genes (GenBank GI numbers 28379168 (SEQ ID NO: 172) and 28377658 (SEQ ID NO: 1111), respectively) were obtained from Lactobacillus plantarum (ATCC No. BAA-793) via polymerase chain reaction (PCR) using primers N1039 and N1040 (for xpk1) and N1041 and N1042 (for eutD). The primer sequences of N1039, N1040, N1041 and N1042 correspond to SEQ ID Nos. 639-642, respectively.
(262) The xpk1 and eutD genes were fused to a DNA fragment containing opposing yeast terminator sequences (CYC and ADH terminators, obtained from PacI digestion of pRS423::CUP1-alsS+FBA-budA, described in U.S. Patent Application Publication No. 20090155870, herein incorporated by reference) by overlap PCR method (Yu et al., Fungal Genet. Biol. 41: 973-981; 2003). The resulting PCR product was cloned into an E. coli-yeast shuttle vector using gap repair methodology (Ma et al., Genetics 58:201-216; 1981). The shuttle vector was based on pRS426 (ATCC No. 77107) and contained both GPD (also known as TDH3) and ADH1 promoters. The resulting vector contained xpk1 under control of the GPD promoter and eutD under control of the ADH1 promoter in opposing orientation. The sequence of the resulting vector (pRS426::GPD-xpk1+ADH1-eutD) is provided as SEQ ID No: 643 (see
Example 2
Construction of Phosphoketolase/Phosphotransacetylase Integration Vector
(263) An expression cassette of the pRS426::GPD-xpk1+ADH1-eutD vector (GPD-xpk1+ADH1-eutD) was prepared by digestion with EcoRI and SacI restriction enzymes. The resulting cassette was ligated into the yeast integration vector pUC19-URA3-MCS which was also prepared by digestion with EcoRI and SacI restriction enzymes.
(264) Vector pUC19-URA3MCS is pUC19-based and contains the sequence of the URA3 gene from Saccaromyces cerevisiae situated within a multiple cloning site (MCS). pUC19 (American Type Culture Collection, Manassas, Va.; ATCC#37254) contains the pMB 1 replicon and a gene coding for beta-lactamase for replication and selection in Escherichia coli. In addition to the coding sequence for LRA3, the sequences from upstream and downstream of this gone are included for expression of the URA3 gene in yeast. The vector can be used for cloning purposes and can be used as a yeast integration vector.
(265) The DNA encompassing the URA3 coding region along with 250 bp upstream and 150 bp downstream of the URA3 coding region from Saccaromyces cerevisiae CEN.PK 113-7D (CBS 8340; Centraalbureau voor Schimmelcultures (CBS) Fungal Biodiversity Centre, Netherlands) genomic DNA was amplified with primers oBP438 (SEQ ID NO: 644), containing BamHI, AscI, Pinel, and FseI restriction sites, and oBP439 (SEQ ID NO: 645), containing XbaI, P acI, and NotI restriction sites. Genomic DNA was prepared using a Gentra Puregene Yeast/Bact kit (Qiagen). The PCR product and, pUC19 were ligated with T4 DNA ligase after digestion with BamHI and XbaI to create vector pUC19-URA3MCS. The vector was confirmed by PCR and sequencing with primers oBP264 (SEQ ID NO:646) and oBP265 (SEQ ID NO:647).
(266) The ligation reaction was transformed into E. coli Stb13 cells, according to the manufacturer's instructions (Invitrogen, Carlsbad, Calif., Cat. No. C7373). Transformants were screened by polymerase chain reaction (PCR) to detect the eutD gene using the primers N1041 and N1042 (SEQ ID NOs: 641 and 642, respectively). Positive clones for eutD gene expression detected by PCR were further confirmed for eutD gene incorporation by digestion of the vector with SacII restriction enzyme.
(267) Two confirmed clones were selected and an integration targeting sequence was added to the clones as follows. PCR was used to amplify regions of the genome of S. cerevisiae strain BY4700 (ATCC No. 200866) both 5 and 3 of the PDC1 gene using the following primers: N1049 and N1050 (5) and N1047 and N1048 (3) (SEQ ID NOs: 648-651, respectively). Primer N1049 enables the 3 end of the 161-bp PDC1 3 sequence to be fused to the 5 end of the 237 bp PDC1 5 sequence via PCR. This pdc 1 3-5-fusion fragment (368 by in length) was cloned into the pCR11Blunt TOPO vector according to the manufacturer's instructions (Invitrogen, Carlsbad, Calif., Cat. No. K2800).
(268) Transformants were screened by PCR to detect the pdc1 3-5-fusion fragment using primers N1047 and N1050. The pdc1 3-5-fusion fragment was isolated from positive clones and released from the vector by digestion with EcoRI enzyme, and ligated into a pUC19-URA3::GPD-xpk1+ADH-eutD vector that had been linearized by digestion with EcoRI restriction enzyme to generate the phosphoketolase pathway vector. Additionally, the pdc1 3-5-fusion fragment was ligated with pUC19-URA3-MCS digested with EcoRI restriction enzyme to generate the control vector. Both ligation reactions were transformed into E. coli Stb13 cells according to the manufacturer's instructions (Invitrogen, Carlsbad, Calif., Cat. No. C7373). The resulting transformants were screened by PCR to detect the pdc1 3-5-fusion fragment using primers N1047 and N1050. Positive clones containing the pdc1 3-5-fusion fragment were identified and the vectors were digested with either NcoI restriction enzyme (control vector) or BsgI restriction enzyme (phosphoketolase pathway vector) to confirm cloning orientation. One control clone (=pUC19-URA3::pdc1) and one phosphoketolase pathway clone (=pUC19-URA3::pde1::GPD-xpk1+ADH1-eutD; SEQ ID NO: 1898) were selected for integration.
Example 3
Construction of Pyruvate Decarboxylase Knockout (PDC-KO) Yeast Strain Containing Phosphoketolase and Phosphotransacetylase Genes
(269) The control and phosphoketolase pathway vectors described in Example 2 were linearized with AflII restriction enzyme and transformed into strain BP913 (CEN.PK113-7D ura3::loxP his3 pdc6 pdc1::ilvD(Sm) pc-1c5::sadB) to form control and phosphoketolase pathway strains. Strain BP913 is further described in Example 10.
(270) Transformed cells were plated on synthetic complete medium without uracil containing ethanol as the sole carbon source (1% vol/vol) and screened by PCR using primers N238 and oBP264 (SEQ ID Nos. 652 and 646, respectively to confirm integration at the pdc1 locus. Integration at the pdc1::ilvD(Sm) locus resulted in the loss of ilvD(Sm).
Example 4
Introduction of Phosphoketolase and Phosphotransacetylase Allows Growth of PDC-KO Cells without Exogenous Two-Carbon Substrate Supplementation
(271) Pyruvate decarboxylase knockout (PDC-KO) yeast strains are unable to grow in media containing 2% glucose as the sole carbon source, but can grow in 2% glucose supplemented with ethanol as shown with a strain transformed with one or more plasmids encoding members of the butanediol pathway (described in U.S. Patent Application Publication No. 20090305363, herein incorporated by reference). To test whether the introduction of the phosphoketolase and phosphotransacetylase genes could support growth of PDC-KO cells, PDC-KO yeast were transformed with the phosphoketolase and phosphotransacetylase gene (as described in Example 3) and with the vector pRS423CUP1-alsS+F13A-budA (described in U.S. Patent Application Publication No. 20090155870, herein incorporated by reference) encoding members of the butanediol pathway. After cultivation in media containing 2% glucose (synthetic complete minus his and ura) supplemented with 0.05% v/v ethanol, cultures were diluted into the same media lacking ethanol (starting OD=0.1, 20 ml medium in a 125 ml vented flask). For comparison, a control PDC-KO strain without introduction of the phosphoketolase and phosphotransacetylase genes was also diluted into medium supplemented with ethanol (0.05% vol/vol). The optical density at 600 nm was measured during growth (results shown in
(272) TABLE-US-00010 TABLE 13 Condition Strain 0 h OD 16 h OD 22 h OD 41.3 h OD xpk A 0.1 2.07 5.63 9.64 xpk B 0.1 2.44 5.93 9.78 xpkC 0.1 2.26 5.83 9.96 control A 0.1 0.47 0.5 0.822 control B 0.1 0.45 0.51 0.849 control C 0.1 0.5 0.52 0.879 cont A w/EtOH 0.1 2.01 5.49 11.44 cont B w/EtOH 0.1 2.16 5.7 11.5 cont C w/EtOH 0.1 2.12 5.76 11.76
(273) The growth of PDC-KO yeast transformed with phosphoketolase and phosphotransacetylase in media that was not supplemented with ethanol (xpkA-xpkC, representing n=3 results) was indistinguishable from the growth of PDC-KO yeast strains grown in media containing 2% glucose that was supplemented with ethanol (cont A-cont C w/EtOH, representing n=3 results). The average growth rate of the phosphoketolase- and phosphotransacetylase-transformed strains under these conditions was 0.19 h.sup.1, A growth rate of 0.23 h.sup.1 for the phosphoketolase- and phosphotransacetylase-transformed strains was observed upon culturing under the same conditions with higher aeration (data not shown). PDC-KO yeast strains grown in media containing 2% glucose that was not supplemented with ethanol showed some growth in the first 16 hours, but then grew at a rate of only 0.01 h.sup.1 (control A-control C, representing n=3 results).
Example 5
Construction of Pyruvate Decarboxylase Knockout (PDC-KO) Yeast Strains Containing Either Phosphoketolase or Phosphotransacetylase Genes
(274) The integration vector described above (pUC19-URA3::pdc1::GPD-xpk1+ADH1-eutD) was modified to eliminate either the xpk1 phosphoketolase gene or the eutD phosphotransacetylase gene. Specifically, to remove eutD, the integration vector was digested with the ClaI and SpeI restriction enzymes to remove a 0.6 kb region from the eutD coding sequence, forming the vector pUC19-URA3::pdc1::GPD-xpk1. To remove xpk1, the integration, vector was digested with the SpeI and KpnI restriction enzymes to remove the 3.4 kb region from SpeI to KpnI, forming the vector pUC19-URA3::pdc1::ADH-eutD. The resulting vectors, were linearized with digestion with the AflII restriction enzyme and transformed into BP913/pRS423::CUP1-alsS+FBA-budA cells (described in Example 3). Transformed cells were screened by PCR to confirm integration at the pdc1 locus and cultured, as described above.
Example 6
Introduction of Phosphoketolase Allows Growth of PDC-KO Cells without Exogenous Two Carbon Substrate Supplementation
(275) To test whether the introduction of either the phosphoketolase or phosphotransacetylase genes could support the growth of PDC-KO cells, PDC-KO yeast were transformed with either the phosphoketolase or phosphotransacetylase genes (as described in Example 5) and with the vector pRS423::CUP1-alsS+FBA-budA encoding members of the butanediol pathway (as described in Example 4). After cultivation in media containing 2% glucose (synthetic complete minus his and ura) supplemented with 0.05% v/v ethanol, cultures were diluted into the same media lacking ethanol (starting OD=0.1, 20 ml medium in a 125 ml vented flask). For comparison, a PDC-KO strain without introduction of the phosphoketolase or phosphotransacetylase genes were grown under the same conditions. The optical density at 600 nm was measured during growth (results shown in
(276) TABLE-US-00011 TABLE 15 Strain 0 h OD 24 h OD xpk1 + eutD 0.1 7.48 none (control) 0.1 0.575 eutD only 0.1 0.338 eutD only 0.1 0.28 xpk1 only 0.1 6.74 xpk1 only 0.1 7.26
(277) The growth of PDC-KO yeast transformed with phosphoketolase in media that was not supplemented with exogenous carbon substrate (xpk1,
Example 7
Introduction of Phosphoketolase to PDC-KO Cells Increases Glucose Consumption and Butanediol Yield
(278) To test the effects of introduction of p iosphoketolase into PDC-KO cells on glucose consumption and butanediol yield, PDC-KO yeast ransformed with either (1) phosphoketolase and phosphotransacetylase (as described in Example 4) and the vector pRS423::CUP1-alsS+FBA-budA encoding members of the butanediol (BDO pathway (as described in Example 4 ) (Xpk in Table 16 below); or with (2) the vector pRS423::CUP1-alsS+FBA-budA encoding members of the butanediol pathway (Control in Table 6 below).
(279) After cultivation in medium containing 2% glucose (synthetic complete minus histidine and uracil) supplemented with 0.05% ethanol, Xpk and Control cultures were diluted into medium without ethanol (starting OD=0.1, 20 ml, medium in a 125 ml vented flask). Glucose consumption and butanediol yield of Xpk and Control cultures were measured by HPLC analysis of culture media for amount of glucose and butanediol as shown in the Table below.
(280) TABLE-US-00012 TABLE 16 Introduction of Phosphoketolase Increases Glucose Consumption and Butanediol Yield of PDC-KO Cells. Glucose consumed Butanediol Strains (mM) Molar Yield Xpk (n = 3) 73.9 2.4 0.475 0.001 Control (n = 3) 48.3 0.6 0.359 0.003
(281) The glucose consumption of Xpk cells (n=3) was nearly twice the amount of glucose consumption of control strains (n=3). In addition, the butanediol molar yield of Xpk cells was increased compared to the butanediol molar yield of Control cells.
Example 8
Construction of an Additional Phosphoketolase Pathway Integration Vector
(282) A phosphoketolase/phosphotransacetylase integration vector similar to the one described in Example 2 was constructed. In this case the xpk1 and eutD gene constricts were cloned so that they would be integrated immediately downstream of the pdc1::ilvD(Sm) locus of BP913. To do this, the intergenic region between ilvD(Sm) and TRX1 was amplified from BP913 genomic DNA using primers N1110 and N1111 (SEQ ID Nos. 653 and 654). This was cloned into pUC19-URA3-MCS at the Pmel site, as follows. The ilvD-TRX1 PCR product was phosphorylated with polynucleotide kinase (NEB Cat. No. MO201), the vector was prepared by digesting with PmeI and treating with calf intestinal phosphatase, the two fragments were ligated overnight and cloned into E. coli Stb13 cells. Clones were screened by PCR (using N1110 and N1111 primers) and then digested with BsgI to determine the orientation of the ilvD-TRX1 insertion. One clone from each orientation (pUC19-URA3::ilvD-TRX1 A and B was carried over to the next step: addition of the xpk1/eutD expression cassette. The xpk1/eutD expression cassette from pRS426::GPD-xpk1+ADH1-eutD was obtained by digestion with BglII and EcoRV. The 5 overhanging DNA was filled in using Klenow Fragment. pUC19-URA3::ilvD-TRX1 was linearized with AflII and the 5 overhanging DNA was filled in using Klenow fragment. This vector was then ligated with the prepared xpk1/eutD cassette. Ligation reactions were transformed into E. coli Stb13 cells. Clones were screened using primers for eutD (N1041 and N1042) and then digested with BamHI to determine orientation of the xpk1/eutD cassette relative to the ilvD-TRX1 DNA sequence.
(283) The URA3 marker gene was then replaced with a geneticin resistance marker as follows. A chimeric geneticin resistance gene was constructed that contained the Kluyveromyces lactis TEF1 promoter and terminator (TEF1p-kan-TEF1t gene, provided as SEQ ID No. 655). This gene was maintained in a pUC19 vector (cloned at the SmaI site). The kan gene was isolated from pUC19 by first digesting with KpnI, removal of 3 overhanging DNA using Klenow Fragment (NEB, Cat. No. M212), digesting, with HincII and then gel purifying the 1.8 kb gene fragment (Zymoclean Gel DNA Recovery Kit, Cat. No. D4001, Zymo Research, Orange, Calif.). The URA3 marker was removed from pUC19-URA3::ilvD::GPD-xpk1+ADH1-eutD::TRX1 (paragraph above) using NsiI and NaeI (the 3 overhanging DNA from NsiI digestion was removed with Klenow fragment). The vector and kan gene were ligated overnight and transformed into E. coli Stb13 cells. Clones were screened by PCR using primers BK468 and either N1090 or N1113 (SEQ ID Nos. 656, 657, and 658, respectively)positive PCR results indicate presence and orientation of kan gene. Clones in both orientations were digested with PmeI and transformed into BP913 with selection on yeast extract-peptone medium supplied with 1% (v/v) ethanol as carbon source and 200 g/ml geneticin (G418). A single transformant was obtained, as confirmed by PCR (primers N886 and oBP264 for the 5 end N1090 and oBP512 for the 3 end, SEQ ID Nos.659, 646, 657, and 660, respectively).
Example 9
Construction of an Isobutanol-Producing Strain Carrying the Phosphoketolase Pathway
(284) The strain described in Example 8 was transformed with 2 plasmids containing genes for an isobutanol pathway pYZ090 and pYZ067 (SEQ ID NOs: 1892 and 1891).
(285) pYZ090 was constructed to contain a chimeric gene having the coding region of the alsS gene from Bacillus subtilis (nt position 457-2172) expressed from the yeast CUP1 promoter (nt 2-449) and followed by the CYC1 terminator (nt 2181-2430) for expression of ALS, and a chimeric gene having the coding region of the ilvC gene from Lactococcus lactis (nt 3634-4656) expressed from the yeast ILV5 promoter (2433-3626) and followed by the ILV5 terminator (nt 4682-5304) for expression of KARI. pYZ067 was constructed to contain the following chimeric genes: 1) the coding region of the ilvD gene from S. mutans UA159 with a C-terminal Lumio tag (nt 2260-3972) expressed from the yeast FBA1 promoter (nt 1661-2250) followed by the FBA1 terminator (nt 40005-4317) for expression of dihydroxy acid dehydratase, 2) the coding region for horse liver ADH (nt 4680-5807) expressed from the yeast GPM1 promoter (nt 5819-6575) followed by the ADH1 terminator (nt 4356-4671) for expression of alcohol dehydrogenase, and 3) the coding region of the kivD gene from Lactococcus lactis (nt 7175-8821) expressed from the yeast TDH3 promoter (nt 8830-9493) followed by the TDH3 terminator (nt 6582-7161) for expression of ketoisovalerate decarboxylase.
(286) Transformants were obtained on synthetic complete medium lacking uracil and histidine with 1% (v/v) ethanol as carbon source and 100 g/ml geneticin. Control strains (BP913) were also transformed with the same plasmids and plated without geneticin. A number of transformants were then patched to the same medium containing 2% glucose as carbon source and supplemented with 0.05% (v/v) ethanol. After 36 hours, patches were used to inoculate liquid medium (same composition as the plates). After 48 hours, ODs for both phosphoketolase pathway and control strains were similar (ca. 4-5 OD) and all were subcultured into medium lacking ethanol (i.e. no exogenous two-carbon substrate source). The phosphoketolase cultures grew without ethanol supplementation, similar to ethanol supplemented control strains. Results are shown in
(287) TABLE-US-00013 TABLE 17A Condition Strain 0 h OD 18.3 h OD xpk ISO 1 0.1 2.3 xpk ISO 2 0.1 2.2 xpk ISO 3 0.1 2.2 ISO (no EtOH) 1 0.1 0.48 ISO (no EtOH) 2 0.1 0.41 ISO (no EtOH) 3 0.1 0.47 ISO (+EtOH) 1 0.1 2.5 ISO (+EtOH) 2 0.1 2.6 ISO (+EtOH) 3 0.1 2.4
(288) TABLE-US-00014 TABLE 17B Condition 0 h 6.5 h 23 h 27 h 48 h Strain OD OD OD OD OD xpk ISO 1 0.1 0.18 2.91 4 4.4 xpk ISO 2 0.1 0.14 1.54 2.88 4.6 xpk ISO 3 0.1 0.16 2.15 3.54 4.3 ISO (+EtOH) 1 0.1 0.14 2.21 3.46 4.4 ISO (+EtOH) 2 0.1 0.11 1.13 2.18 4.2 ISO (+EtOH) 3 0.1 0.1 0.84 1.6 4.4
Example 10
Construction of Saccharomyces cerevisiae Strain BP913
(289) The purpose of this example is to describe the construction of Saccharomyces cerevisiae strain BP913. The strain was derived from CEN.PK 113-7D (CBS 8340; Centraalbureau voor Schimmelcultures (CBS) Fungal Biodiversity Centre, Netherlands) and contains deletions of the following genes: URA3, HIS3, PDC1, PDC5, and PDC6.
(290) Deletions, which completely removed the entire coding sequence, were created by homologous recombination with PCR fragments containing regions of homology upstream and downstream of the target gene and either a G418 resistance marker or URA3 gene for selection of transformants. The G418 resistance marker, flanked by loxP sites, was removed using Cre recombinase. The URA3 gene was removed by homologous recombination to create a scarless deletion.
(291) In general, the PCR cassette for each scarless deletion was made by combining four fragments, A-B-U-C, by overlapping PCR. The PCR cassette contained a selectable/counter-selectable marker, URA3 (Fragment U), consisting of the native CEN.PK 113-7D URA3 gene, along with the promoter (250 bp upstream of the URA3 gene) and terminator (150 bp downstream of the URA3 gene). Fragments A and C, each 500 bp long, corresponded to the 500 bp immediately upstream of the target gene (Fragment A) and the 3 500 by of the target gene (Fragment C). Fragments A and C were used for integration of the cassette into the chromosome by homologous recombination. Fragment B (500 by long) corresponded to the 500 bp immediately downstream of the target gene and was used for excision of the URA3 marker and Fragment C from the chromosome by homologous recombination, as a direct repeat of the sequence corresponding to Fragment B was created upon integration of the cassette into the chromosome. Using the PCR product ABUC cassette, the URA3 marker was first integrated into and then excised from the chromosome by homologous recombination. The initial integration deleted the gene, excluding the 3 500 bp. Upon excision, the 3 500 bp region of the gene was also deleted. For integration of genes using this method, the gene to be integrated was included in the PCR cassette between fragments A and B. URA3 Deletion
(292) To delete the endogenous URA3 coding region, a ura3::loxP-kanMX-loxP cassette was PCR-amplified from pLA54 template DNA (SEQ ID NO: 661). pLA54 contains the K. lactis TEF1 promoter ar d kanMX marker, and is flanked by loxP sites to allow recombination with Cre recombinase and removal of the marker. PCR was done using Phusion DNA polymerase and primers BK505 and BK506 (SEQ ID NOs:662 and 663). The URA3 portion of each primer was derived from the 5 region upstream of the URA3 promoter and 3 region downstream of the coding region such that integration of the loxP-kanMX-loxP marker resulted in replacement of the URA3 coding region. The PCR product was transformed into CEN.PK 113-7D using standard genetic techniques (Methods in Yeast Genetics, 2005, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., pp. 201-202) and transformants were selected on YPD containing G418 (100 g/ml) at 30 C. Transformants were screened to verify correct integration by PCR using primers LA468 and LA492 (SEQ ID NOs: 664 and 665) and designated CEN.PK 113-7D ura3::kanMX.
(293) HIS3 Deletion
(294) The four fragments for the PCR cassette for the scarless HIS3 deletion were amplified using Phusion High Fidelity PCR Master Mix (New England BioLabs) and CEN.PK 113-7D genomic DNA as template, prepared with a Centra Puregene Yeast/Bact kit (Qiagen; Valencia, Calif.). HIS3 Fragment A was amplified with primer oBP452 (SEQ ID NO: 666) and primer oBP453 (SEQ ID NO: 667), containing a 5 tail with homology to the 5 end of HIS3 Fragment B. HIS3 Fragmeht B was amplified with primer oBP454 (SEQ ID NO: 668), containing a 5 tail with homology to the 3 end of HIS3 Fragment A, and primer oBP455 (SEQ ID NO: 669), containing a 5 tail with homology to the 5 end of HIS3 Fragment U. HIS3 Fragment U was amplified with primer oBP456 (SEQ ID NO: 670), containing a 5 tail with homology to the 3 end of HIS3 Fragment B, and primer oBP457 (SEQ ID NO: 671), containing a 5 tail with homology to the 5 end of HIS3 Fragment C. HIS3 Fragment C was amplified with primer oBP458 (SEQ ID NO: 672), containing a 5 tail with homology to the 3 end of HIS3 Fragment U, and primer oBP459 (SEQ ID NO: 673). PCR products were purified with a PCR Purification kit (Qiagen, Valencia, Calif.). HIS3 Fragment AB was created by overlapping PCR by mixing HIS3 Fragment A and HIS3 Fragment B and amplifying with primers oBP452 (SEQ ID NO: 666) and oBP455 (SEQ ID NO: 669). HIS3 Fragment UC was created by overlapping PCR by mixing HIS3 Fragment U and HIS3 Fragment C and amplifying with primers oBP456 (SEQ ID NO: 670) and oBP459 (SEQ ID NO: 673). The resulting PCR products were purified on an agarose gel followed by a Gel Extraction kit (Qiagen, Valencia, Calif.). The HIS3 ABUC cassette was created by overlapping PCR by mixing HIS3 Fragment AB and HIS3 Fragment UC and amplifying with primers oBP452 (SEQ ID NO: 666) and oBP459 (SEQ ID NO: 673). The PCR product was purified with a PCR Purification kit (Qiagen, Valencia, Calif.).
(295) Competent cells of CEN.PK 113-7D) ura3::kanMX were made and transformed with the HIS3 ABUC PCR cassette using a Frozen-EZ Yeast Transformation II kit (Zymo Research, Orange, Calif.). Transformation mixtures were plated on synthetic complete media lacking uracil supplemented with 2% glucose at 30 C. Transformants with a his knockout were screened for by PCR with primers oBP460 (SEQ ID NO: 674) and oBP461 (SEQ ID NO: 671) using genomic DNA prepared with a Gentra Puregene Yeast/Bact kit (Qiagen). A correct transformant was selected as strain CEN.PK 113-7D ura3::kanMX his3::URA3.
(296) KanMX Marker Removal from the ura3 Site and URA3 Marker Removal from the his3 Site
(297) The KanMX marker was removed by transforming CEN.PK 113-7D ura3::kanMX his3::URA3 with pRS423::PGAL1-cre (SEQ ID NO: 715) using a Frozen-EZ Yeast Transformation II kit (Zymo Research, Orange, Calif.) and plating on synthetic complete medium lacking histidine and uracil supplemented with 2% glucose at 30 C. Transformants were grown in YP supplemented with 1% galactose at 30 C for 6 hours to induce the Cre recombinase and KanMX marker excision and plated onto YPD (2% glucose) plates at 30 C for recovery. An isolate was grown overnight in YPD and plated on synthetic complete medium containing 5-fluoro-orotic acid (0.1%) at 30 C to select for isolates that lost the URA3 marker. 5-FOA resistant isolates were grown in and plated on YPD for removal of the pRS423::PGAL1-cre plasmid. Isolates were checked for loss of the KanMX marker, URA3 marker, and pRS423::PGAL1-cre plasmid by assaying, growth on YPD+G418 plates, synthetic complete medium lacking uracil plates, and synthetic complete medium lacking histidine plates. A correct isolate that was sensitive to G418 and auxotrophic for uracil and histidine was selected as strain CEN.PK 113-7D ura3::loxP his3 and designated as BP857. The deletions and marker removal were confirmed by PCR and sequencing with primers oBP450 (SEQ ID NO: 676) and oBP451 (SEQ ID NO: 677) for ura3 and primers oBP460 (SEQ ID NO: 674) and oBP461 (SEQ ID NO: 675) for his3 using genomic DNA prepared with a Gentra Puregene Yeast/Bact kit (Qiagen, Valencia, Calif.).
(298) PDC6 Deletion
(299) The four fragments for the PCR cassette for the starless PDC6 deletion were amplified using Phusion High Fidelity PCR Master Mix (New England BioLabs, Ipswich, Mass.) and CEN.PK 113-7D genomic DNA as template, prepared with a Gentra Puregene Yeast/Bact kit (Qiagen). PDC6 Fragment A was amplified with primer oBP440 (SEQ ID NO: 670) and primer oBP441 (SEQ ID NO: 679), containing a 5 tail with homology to the 5 end of PDC6 Fragment B. PDC6 Fragment B was amplified with primer oBP442 (SEQ ID NO: 680), containing a 5 tail with homology to the 3 end of PDC6 Fragment A, and primer oBP443 (SEQ ID NO: 681), containing, a 5 tail with homology to the 5 end of PDC6 Fragment U. PDC6 Fragment U was amplified with primer oBP444 (SEQ ID NO: 682), containing a 5 tail with homology to the 3 end of PDC6 Fragment B, and primer oBP445 (SEQ ID NO: 683), containing a 5 tail with homology to the 5 end of PDC6 Fragment C. PDC6 Fragment C was amplified with primer oBP446 (SEQ ID NO: 684), containing a 5 tail with homology to the 3 end of PDC6 Fragment U, and primer oBP447 (SEQ ID NO: 685). PCR products were purified with, a PCR Purification kit (Qiagen). PDC6 Fragment AB was created by overlapping PCR by mixing PDC6 Fragment A and PDC6 Fragment B and amplifying with primers oBP440 (SEQ ID NO: 678) and oBP443 (SEQ ID NO: 681). PDC6 Fragment UC was created by overlapping PCR by mixing PDC6 Fragment U and PDC6 Fragment C and amplifying with primers oBP444 (SEQ ID NO: 682) and oBP447 (SEQ ID NO: 685). The resulting PCR products were purified on an agarose gel followed by a Gel Extraction kit (Qiagen). The PDC6 ABUC cassette was created by overlapping PCR by mixing PDC6 Fragment AB and PDC6 Fragment UC and amplifying with primers oBP440 (SEQ ID NO: 678) and oBP447 (SEQ ID NO 685). The PCR product was purified with a PCR Purification kit (Qiagen).
(300) Competent cells of CEN.PK 113-7D ura3::loxP his3 were made and transformed with the PDC6 ABUC PCR cassette using a Frozen-EZ Yeast Transformation II kit (Zymo Research), Transformation mixtures were plated on synthetic complete media lacking uracil supplemented with 2% glucose at 30 C. Transformants with a pdc6 knockout were screened for by PCR with primers oBP448 (SEQ ID NO 686) and oBP449 (SEQ ID NO; 687) using genomic DNA prepared with a Gentra Puregene Yeast/Bact kit (Qiagen). A correct transformant was selected as strain CEN.PK 113-7D ura3::loxP his3 pdc6::URA3.
(301) CEN.PK 113-7D ura3::loxP his3 pdc6::URA3 was grown overnight in YPD and plated on synthetic complete medium containing 5-fluoro-orotic acid (0,1%) at 30 C to select for isolates that lost the URA3 marker. The deletion and marker removal were confirmed by PCR and sequencing with primers oBP448 (SEQ ID NO: 686) and oBP449 (SEQ ID NO 687) using genomic DNA prepared with a Gentra Puregene Yeast/Bact kit (Qiagen). The absence of the PDC6 gene from the isolate was demonstrated by a negative PCR result using primers specific for the coding sequence of PDC6, oBP554 (SEQ ID NO 688) and oBP555 (SEQ ID NO 689). The correct isolate was selected as strain CEN.PK 113-7D ura3::loxP his3 pdc6 and designated as BP891.
(302) PDC1 Deletion ilvDSm Integration
(303) The PDC1 gene was deleted and replaced with the ilvD coding region from Streptococcus mutans ATCC #700610 (SEQ ID NO: 1886). The A fragment followed by the ilvD coding region from Streptococcus mutans for the PCR cassette for the PDC1 deletion-ilvDSm integration was amplified using Phusion High Fidelity PCR Master Mix (New England BioLabs) and NYLA83 genomic DNA as template (construction of strain NYLA83 is described in U.S. Application Pub. No. 20110124060 A1, incorporated herein by reference), prepared with a Gentra Puregene Yeast/Bact kit (Qiagen). PDC1 Fragment A-ilvDSm was amplified with primer oBP513 (SEQ ID NO: 690) and primer oBP515 (SEQ ID NO: 691), containing a 5 tail with homology to the 5 end of PDC1 Fragment B. The B, U, and C fragments for the PCR cassette for the PDC1 deletion-ilvDSm integration were amplified using Phusion High Fidelity PCR Master Mix (New England BioLabs) and CEN.PK 113-7D genomic DNA as template, prepared with a Gentra Puregene Yeast/Bact kit (Qiagen). PDC1 Fragment B was amplified with primer oBP516 (SEQ ID NO: 692), containing a 5 tail with homology to the 3 end of PDC1 Fragment A-ilvDSm, and primer oBP517 (SEQ ID NO: 693), containing a 5 tail with homology to the 5 end of PDC1 Fragment U. PDC1 Fragment U was amplified with primer oBP518 (SEQ ID NO: 694), containing a 5 tail with homology to the 3 end of PDC1 Fragment B, and primer oBP519 (SEQ ID NO: 695), containing a 5 tail with homology to the 5 end of PDC1 Fragment C. PDC1 Fragment C was amplified with primer oBP520 (SEQ ID NO: 696), containing a 5 tail with homology to the 3 end of PDC1 Fragment U, and primer oBP521 (SEQ ID NO: 697). PCR products were purified with a PCR Purification kit (Qiagen). PDC1 Fragment A-ilvDSm-B was created by overlapping PCR by mixing PDC1 Fragment A-ilvDSrn and PDC1 Fragment B and amplifying with primers oBP513 (SEQ ID NO: 690) and oBP517 (SEQ ID NO: 693). PDC Fragment UC was created by overlapping PCR by mixing PDC1 Fragment U and PDC1 Fragment C and amplifying with primers oBP518 (SEQ ID NO: 694) and oBP521 (SEQ ID NO: 697). The resulting PCR products were purified on an agarose gel followed by a Gel Extraction kit (Qiagen). The PDC 1 A-ilvDSm-BUC cassette was created by overlapping PCR by mixing PDC 1 Fragment A-ilvDSm-B and PDC1 Fragment UC and amplifying with primers oBP513 (SEQ ID NO 690) and oBP521 (SEQ ID NO: 697). The PCR product was purified with a PCR Purification kit (Qiagen).
(304) Competent cells of CEN.PK 113-7D ura3::loxP his3 pdc6 were made and transformed with the PDC1 A-ilvDSm-BUC PCR cassette using a Frozen-EZ Yeast Transformation II kit (Zymo Research). Transformation mixtures were plated on synthetic complete media lacking uracil supplemented with 2% glucose at 30 C. Transformants with a pdc1 knockout ilvDSm integration were screened for by PCR with primers oBP511 (SEQ ID NO: 698) and oBP512 (SEQ ID NO: 699) using genomic DNA prepared with a Gentra Puregene Yeast/Bact kit (Qiagen). The absence of the PDC1 gene from the isolate was demonstrated by a negative PCR result using primers specific for the coding sequence of PDC1, oBP550 (SEQ ID NO: 700) and oBP551 (SEQ ID NO: 701). A correct transformant was selected as strain CEN.PK 113-7D ura3::loxP his3 pdc6 pdc 1::ilvDSm-URA3.
(305) CEN.PK 113-7D ura3::loxP his3 pdc6 pdc1::ilvDSm-URA3 was grown overnight in YPD and plated on synthetic complete medium containing 5-fluoro-orotic acid (0.1%) at 30 C to select for isolates that lost the URA3 marker. The deletion of PDC1, integration of ilvDSm, and marker removal were confirmed by PCR and sequencing with primers oBP511 (SEQ ID NO: 698) and oBP512 (SEQ ID NO: 699) using genomic DNA prepared with a Gentra Puregene Yeast/Bact kit (Qiagen). The correct isolate was selected as strain CEN.PK 113-7D ura3::loxP his3 pdc6 pdc1::ilvDSm and designated as BP907.
(306) PDC5 Deletion sadB Integration
(307) The PDC5 gene was deleted and replaced with the sadB coding region from Achromobacter xylosoxidans. A segment of the PCR cassette for the PDC5 deletion-sadB integration was first cloned into plasmid pUC19-URA3MCS (described in Example 2). The coding sequence of sadB (SEQ ID NO: 718) and PDC5 Fragment B were cloned into pUC19-URA3MCS to create the sadB-BU portion of the PDC5 A-sadB-BUC PCR cassette. The coding sequence of sadB was amplified using pLH468-sadB (SEQ ID NO: 716) as template with primer oBP530 (SEQ ID NO: 702), containing an AscI restriction site, and primer oBP531 (SEQ ID NO: 703), containing a 5 tail with homology to the 5 end of PDC5 Fragment B. PDC5 Fragment B was amplified from CEN.PK 113-7D genomic DNA with primer oBP532 (SEQ ID NO: 704), containing a 5 tail with homology to the 3 end of sadB, and primer oBP533 (SEQ ID NO: 705), containing a PmeI restriction site PCR products were purified with a PCR Purification kit (Qiagen). sadB-PDC5 Fragment B was created by overlapping PCR by mixing the sadB and PDC5 Fragment B PCR products and amplifying with primers oBP530 (SEQ ID NO: 702) and oBP533 (SEQ ID NO 705). The resulting PCR product was digested with Asci and PmeI and ligated with T4 DNA ligase into the corresponding sites of pUC19-URA3MCS after digestion with the appropriate enzymes. The resulting plasmid was used as a template for amplification of sadB-Fragment B-Fragment U using primers oBP536 (SEQ ID NO: 706) and oBP546 (SEQ ID NO: 707), containing a 5 tail with homology to the 5 end of PDC5 Fragment C. PDC5 Fragment C was amplified from CEN.PK 113-7D genomic DNA with primer oBP547 (SEQ ID NO: 708), containing a 5 tail with homology to the 3 end of PDC5 sadB-Fragment B-Fragment U, and primer oBP539 (SEQ ID NO: 709). PCR products were purified with a PCR Purification kit (Qiagen). PDC5 sadB-Fragment B-Fragment U-Fragment C was created by overlapping PCR by mixing PDC5 sadB-Fragment B-Fragment U and PDC5 Fragment C and amplifying with primers oBP536 (SEQ ID NO: 706) and oBP539 (SEQ II) NO: 709). The resulting PCR product was purified on an agarose gel followed by a Gel Extraction kit (Qiagen). The PDC5 A-sadB-BUC cassette was created by amplifying PDC5 sadB-Fragment B-Fragment U-Fragment C with primers oBP542 (SEQ ID NO: 710) containing a 5 tail with homology to the 50 nucleotides immediately upstream of the native PDC5 coding sequence, and oBP539 (SEQ ID NO: 709). The PCR product was purified with a PCR Purification kit (Qiagen).
(308) Competent cells of CEN.PK 113-7D ura3::loxP his3 pdc6 pdc1::ilvDSm were made and transformed with the PDC5 A-sadB-13UC PCR cassette using a Frozen-EZ Yeast Transformation II kit (Zymo Research). Transformation mixtures were plated on synthetic complete media lacking uracil supplemented with 1% ethanol (no glucose) at 30 C. Transformants with a pdc5 knockout sadB integration were screened for by PCR with primers oBP540 (SEQ ID NO: 711) and oBP541 (SEQ ID NO: 712) using genomic DNA prepared with a Gentra Puregene Yeast/Bact kit (Qiagen). The absence of the PDC5 gene from the isolate was demonstrated by a negative PCR result using primers specific for the coding sequence of PDC5, oBP552 (SEQ ID NO: 713) and oBP553 (SEQ ID NO: 714). A correct transformant was selected as strain CEN.PK 113-7D ura3::loxP his3 pdc6 pdc1::ilvDSrn pdc5::sadB-URA3.
(309) CEN.PK 113-7D ura3::loxP his3 pdc6 pdc1::ilvDSm pdc5::sadB-URA3 was grown overnight in YPE (1% ethanol) and plated on synthetic complete medium supplemented with ethanol (no glucose) and containing 5-fluoro-orotic acid (0.1%) at 30 C to select for isolates that lost the URA3 marker. The deletion, of PDC5, integration of sadB, and marker removal were confirmed by PCR with primers oBP540 (SEQ ID NO 711) and oBP541 (SEQ ID NO: 712) using genomic DNA prepared with a Gentra Puregene Yeast/Bact kit (Qiagen). The correct isolate was selected as strain CEN.PK 113-7D ura3::loxP his3 pdc6 pdc1::ilvDSm pdc5::sadB and designated as BP913.
Example 11
Construction of Strain NYLA83
(310) This example describes insertion-inactivation of endogenous PDC1 and PDC6 genes of S. cerevisiae. PDC1, PDC5, and PDC6 genes encode the three major isozymes of pyruvate decarboxylase. The resulting strain was used as described in Example 10.
(311) Construction of pRS425::GPM-sadB
(312) A DNA fragment encoding a butanol dehydrogenase (SEQ ID NO: 717) from Achromobacter xylosoxidans (disclosed in US Patent Application Publication No. US20090269823) was cloned. The coding region of this gene called sadB for secondary alcohol dehydrogenase (SEQ ID NO: 718) was amplified using standard conditions from A. xylosoxidans genomic DNA, prepared using a Gentra Puregene kit (Gentra Systems, Inc., Minneapolis, Minn.; catalog number D-5500A) following the recommended protocol for gram negative organisms using forward and reverse primers N473 and N469 (SEQ ID NOs:725 and 726), respectively. The PCR product was TOPO-Blunt cloned into pCR4 BLUNT (Invitrogen) to produce pCR4Blunt::sadB, which was transformed into E. coli Mach-1 cells. Plasmid was subsequently isolated from four clones, and the sequence verified.
(313) The sadB coding region was PCR amplified from pCR4Blunt::sadB. PCR primers contained additional 5 sequences that would overlap with the yeast GPM 1 promoter and the ADH1 terminator (N583 and N584, provided as SEQ ID NOs:727 and 728). The PCR product was then cloned using gap repair methodology in Saccharomyces cerevisiae (Ma et al. ibid) as follows. The yeast-E. coli shuttle vector pRS425::GPM::kivD::ADH which contains the GPM1 promoter (SEQ ID NO:721), kivD coding region from Lactococcus lactis (SEQ D NO:719), and ADH1 terminator (SEQ ID NO:722) (described in U.S. Pat. No. 7,851,188, Example 17) was digested with BbvCI and PacI restriction enzymes to release the kivD coding region. Approximately 1 g of the remaining vector fragment was transformed into S. cerevisiae strain 13Y4741 along, with 1 mg of sadB PCR product. Transformants were selected on synthetic complete medium lacking leucine. The proper recombination event, generating pRS425::GPM-sadB, was confirmed by PCR using primers N142 and N459 (SEQ ID NOs:729 and 730).
(314) Construction of pdc6::P.sub.GPM1-sadB Integration Cassette and PDC6 Deletion:
(315) A pdc6:: P.sub.GPM1-sadB-ADH/t-URA3r integration cassette was made by joining the GPM-sadB-ADHt segment (SEQ ID NO:723) from pRS425::GPM-sadB (SEQ ID NO: 720) to the URA3r gene from pUC19-URA3r. pUC19-URA3r (SEQ ID NO:724) contains the URA3 marker from pRS426 (ATCC #77107) flanked by 75 by homologous repeat sequences to allow homologous recombination in vivo and removal of the URA3 marker. The two DNA segments were joined by SOE PCR (as described by Horton et al. (1989) Gene 77:61-68) using as template pRS425::GPM-sadB and pUC19-URA3r plasmid DNAs, with Phusion DNA polymerase (New England Biolabs Inc., Beverly, Mass.; catalog no. F-5405) and primers 114117-11A through 114117-11D (SEQ ID NOs:731, 732, 733 and 734), and 114117-13A and 114117-13B (SEQ ID NOs:735 and 736).
(316) The outer primers for the SOE PCR (114117-13A and 114117-13B) contained 5 and 3 50 by regions homologous to regions upstream and downstream of the PDC6 promoter and terminator, respectively. The completed cassette PCR fragment was transformed into BY4700 (ATCC #200866) and transformants were maintained on synthetic complete media lacking uracil and supplemented with 2% glucose at 30 C. using standard genetic techniques (Methods in Yeast Genetics, 2005, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., pp. 201-202). Transformants were screened by PCR using primers 112590-34G and 112590-34H (SEQ ID NOs:737 and 738), and 112590-34F and 112590-49E (SEQ ID NOs: 739 and 740) to verify integration at the PDC6 locus with deletion of the PDC6 coding region. The URA3r marker was recycled by plating on synthetic complete media supplemented with 2% glucose and 5-FOA at 30 C. following standard protocols. Marker removal was confirmed by patching colonies from the 5-FOA plates onto SD-URA media to verify the absence of growth. The resulting identified strain has the genotype: BY4700pdc6::P.sub.Gpmi-sadB-ADH1t.
(317) Construction of pdc1::P.sub.PDC1-ilvD Integration Cassette and PDC1 Deletion:
(318) A pdch:: P.sub.PDC1-ilvD-FBA1t-URA3r integration cassette was made by joining the ilvD-FBA1t segment (SEQ ID NO:741) from pLH468 (SEQ ID NO: 1888) to the URA3r gene from pUC19-URA3r by SOE FCR (as described by Horton et al. (1989) Gene 77:61-68) using as template pLH468 and pUC19-UR.\3r plasmid DNAs, with Phusion DNA polymerase (New England Biolabs Inc., Beverly, Mass.; catalog no. F-540S) and primers 114117-27A through 114117-27D (SEQ ID NOs:742, 743, 744 and 745).
(319) The outer primers for the SOE PCR (114117-27A and 114117-27D) contained 5 and 3 50 by regions homologous to regions downstream of the PDC1 promoter and downstream of the PDC1 coding sequence. The completed cassette PCR fragment was transformed into BY4700 pdc6::P.sub.GPM1-sadB-ADH1t and transformants were maintained on synthetic complete media lacking uracil and supplemented with 2% glucose at 30 C. using standard, genetic techniques (Methods in Yeast Genetics, 2005, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., pp. 201-202). Transformants were screened by PCR using primers 114117-36D and 135 (SEQ ID NOs 746 and 747), and primers 112590-49E and 112590-30F (SEQ ID NOs 740 and 748) to verify integration at the PDC1 locus with deletion of the PDC1 coding sequence. The URA3r marker was recycled by plating on synthetic complete media supplemented with 2% glucose and 5-FOA at 30 C. following standard protocols. Marker removal was confirmed by patching colonies from the 5-FOA plates onto SD-UR \ media to verify the absence of growth. The resulting identified strain NYLA67 has the genotype: BY4700 pdc6:: P.sub.GPM1-sadB-ADH1t pdc1::_P.sub.PDC1-ilvD-FBA1t.
(320) HIS3 Deletion
(321) To delete the endogenous HIS3 coding region, a his 3:URA3r2 cassette was PCR-amplified from URA3r2 template DNA (SEQ ID NO: 749). URA3r2 contains the URA3 marker from pRS426 (ATCC #77107) flanked by 500 by homologous repeat sequences to allow homologous recombination in vivo and removal of the URA3 marker. PCR was done using Phusion DNA polymerase and primers 114117-45A and 114117-45B (SEQ ID NOs:750 and 751) which generated a 2.3 kb PCR product. The HIS3 portion of each primer was derived from the 5 region upstream of the HIS3 promoter and 3 region downstream of the coding region such that integration of the URA3r2 marker results in replacement of the HIS3 coding region. The PCR product was transformed into NYLA67 using standard genetic techniques (Methods in Yeast Genetics, 2005, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., pp. 201-202) and transformants were selected on synthetic complete media lacking, uracil and supplemented with 2% glucose at 30 C. Transformants were screened to verify correct integration by replica plating of transformants onto synthetic complete media lacking histidine and supplemented with 2% glucose at 30 C. The URA3r marker was recycled by plating on synthetic complete media supplemented with 2% glucose and 5-FOA at 30 C. following standard protocols. Marker removal was confirmed by patching colonies, from the 5-FOA plates onto SD-URA. media to verify the absence of growth. The resulting identified strain, called NYLA73, has the genotype: BY4700 pdc6 . . . P.sub.GPM1/sadB-ADH1t ppc1:: P.sub.PDC1-ilvD-FBA1t his3.
(322) Construction of pdc5::kanMX Integration Cassette and PDC5 Deletion:
(323) A pdc5::kanMX4 cassette was PCR-amplified from strain YLR134W chromosomal DNA (ATCC No. 4034091) using Phusion DNA polymetase and primers PDC5::KanMXF and PDC5.:KanMXR (SEQ ID NOs:752 and 753) which generated a 2.2 kb PCR product. The PDC5 portion of each primer was derived from the 5 region upstream of the PDC5 promoter and 3 region downstream of the coding region such that integration of the kanMX4 marker results in replacement of the PDC5 coding region. The PCR product was transformed into NYLA73 using standard genetic techniques (Methods in Yeast Genetics, 2005, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., pp. 201-202) and transformants were selected on YP media supplemented with 1% ethanol and geneticin (200 g/ml) at 30 C. Transformants were screened by PCR to verify correct integration at the PDC locus with replacement of the PDC5 coding region using primers PDC5kofor and N175 (SEQ ID NOs: 754 and 755). The identified correct transformants have the genotype: BY4700 pdc 6:: P.sub.GPM1-sadB-ADH1t pdc 1::P.sub.PDC1-ilvD-FBA1t his3 pdc5::kanMX4. The strain was named NYLA74.
(324) Deletion of HXK2 (hexokinase II):
(325) A hxk2::URA3r cassette was PCR-amplified from URA3r2 template (described above) using Phusion DNA polymerase and primers 384 and 385 (SEQ ID NOs:756 and 757) which generated a 2.3 kb PCR product. The HXK2 portion of each primer was derived from the 5 region upstream of the HXK2 promoter and 3 region downstream of the coding region such that integration of the URA3r2 marker results in replacement of the HXK2 coding region. The PCR product was transformed into NYLA73 using standard genetic techniques (Methods in Yeast Genetics, 2005, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., pp. 201-202) and transformants were selected on synthetic complete media lacking uracil and supplemented with 2% glucose at 30 C. Transformants were screened by PCR to verify correct integration at the HXK2 locus with replacement of the HXK2 coding region using primers N869 and N871 (SEQ ID NOs:758 and 759). The URA3r2 marker was recycled by plating on synthetic complete media supplemented with 2% glucose and 5-FOA at 30 C. following standard protocols. Marker removal was confirmed by patching colonies from the 5-FOA plates onto SD-URA media to verify the absence of growth, and by PCR to verify correct marker removal using primers N946 and N947 (SEQ ID NOs:760 and 761). The resulting identified strain named NYLA83 has the genotype: BY4700 pdc6:: P.sub.GPM1-sadB-ADH1t pdc1:: P.sub.PDC1-ilvD-FBA1t his3 hxk2.
Example 12
Construction of Saccharomyces cerevisiae Strain PNY2257
(326) Strain PNY2242 was constructed in several steps from BP913 (described above). First, the native GPD2 gene on Chromsome XV was deleted. The coding region was deleted using CRE-lox mediated marker removal (methodology described above), so the resulting locus contains one loxP site. The sequence of the modified locus is provided as SEQ ID NO 1899 (Upstream region=nt 1-500; loxP site=nt 531-564; Downstream region=nt 616-1115). Second, the native FRA2 gene on Chromosome VII was deleted. Elimination of FRA2 was a scarless deletion of only the coding region. The sequence of the modified locus is provided as SEQ ID NO 1900 (Upstream region=nt 1-501; Downstream region=nt 526-1025). Next, the ADH1 gene on Chromosome XV was deleted along with insertion of a chimeric gene comprised of the UAS(PGK1)FBA1 promoter and the kivD coding region. The native ADH1 terminator was used to complete the gene. The sequence of the modified locus is provided as SEQ ID No. 1901 (Upstream region=nt 1-500; UAS(PGK1)FBA promoter=nt 509-1233; kivD coding region=nt 1242-2888; Downstream region (includes terminator)=nt 2889-3388). Next, a chimeric gene comprised of the FBA1 promoter, the alsS coding region and the CYC1 terminator was integrated into Chromosome XII, upstream of the TRX1 gene. The sequence of the modified locus is provided as SEQ ID No. 1902(Upstream region=nt 1-154; FBA1 promoter=nt 155-802; alsS CDS=nt 810-2525; CYC1 terminator=nt 2534-2788; Downstream region=nt 2790-3015). Next, two copies of a gene encoding, horse liver alcohol dehydrogenase were integrated into Chromsomes VII and XVI. On Chromosome VII, a chimeric gene comprised of the PDC1 promoter, the hADH coding region and the ADH 1 terminator were placed into the fra2locus (the original deletion of FRA2 is described above). The sequence of the modified locus is provided as SEQ ID No. 1903 (Upstream region=nt 1-300; PDC1 promoter=nt 309-1178; hADH coding region=nt 1179-2306; ADH1 terminator=nt 2315-2630; Downstream region=nt 2639-2900). On Chromosome XVI, a chimeric gene comprised of the PDC5 promoter, the hADH coding region and the ADH1 terminator were integrated in, the region formerly occupied by the long term repeat element YPRCdelta15. The sequence of the modified locus is provided as SEQ ID No. 1904 (Upstream region=nt 1-150; PDC5 promoter=nt 159-696; hADH coding region=nt 697-1824; ADH1 terminator=nt 1833-2148: Downstream region=nt 2157-2656). Then the native genes YMR226c and ALD6 were deleted. Elimination of YMR226c was a scarless deletion of only the coding region. The sequence of the modified locus is provided as SEQ ID No. 1905 (Upstream region=nt 1-250; Downstream region=nt 251-451). The ALD6 coding region plus 700 by of upstream sequence were deleted using CRE-lox mediated marker removal, so the resulting locus contains one loxP site. The sequence of the modified locus is provided as SEQ ID No. 1906(Upstream region=nt 1-500; loxP site=nt 551-584; Downstream region=nt 678-1128). The geneticin-selectable phosphoketolase expression vector described in Example 8 was transformed into the strain and confirmed as described above (the locus is depicted in
(327) Growth rates were assessed as described in previous examples. Over a 24 hour period, PNY2257 displayed growth rates without ethanol or other two-carbon supplement similar to those growth rates observed for PNY2242 with supplementation.
(328) TABLE-US-00015 TABLE 6 HMMER2.0 [2.2g] NAME XFP_XPK_exp_seqs LENG 845 ALPH Amino RF no CS no MAP yes COM /app/public/hmmer/current/bin/hmmbuild XFP_XPK_HMM XFP_XPK_exp_seqs.aln COM /app/public/hmmer/current/bin/hmmcalibrate - mean 800 XFP_XPK_HMM NSEQ 8 DATE Fri Dec 4 15:29:49 2009 CKSUM 6589 XT 8455 4 1000 1000 8455 4 8455 4 NULT 4 8455 NULE 595 1558 85 338 294 453 1158 197 249 902 1085 142 21 313 45 531 201 384 1998 644 EVD 614.573792 0.077043 A C D E F G H I K HMM m->m m->i m->d i->m i->i d->m d->d b->m m->e L M N P Q R S T V W Y 476 * 1831 1(M) 579 646 1823 1279 241 1949 816 340 991 1145 2778 1234 1286 828 1131 1025 537 205 1033 696 11 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 25 6428 7470 894 1115 701 1378 476 * 2(W) 424 1143 649 112 1118 1430 73 944 1343 1091 364 285 1428 217 174 480 363 730 3083 756 12 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 25 6428 7470 894 1115 701 1378 * * 3(Q) 411 1812 202 1278 2120 997 53 1844 1011 1806 940 170 1370 1483 39 277 358 1435 1985 1327 13 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 25 6428 7470 894 1115 701 1378 * * 4(M) 554 1770 500 1294 2084 1496 45 1741 1490 1726 2375 163 1571 384 1179 467 467 1393 1906 1358 14 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 19 6836 7878 894 1115 701 1378 * * 5(V) 582 614 1167 546 657 1623 419 47 453 583 188 696 1707 324 741 630 773 1221 1069 649 15 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 19 6836 7878 894 1115 701 1378 * * 6(M) 388 632 233 716 648 1697 471 1170 550 499 1779 218 1768 407 821 698 1278 799 1059 646 16 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7025 8067 894 1115 701 1378 * * 7(D) 54 2137 2020 1380 2436 1385 196 2201 126 2144 1250 1219 1583 239 702 471 581 1750 2320 1600 17 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7025 8067 894 1115 701 1378 * * 8(Y) 508 822 1280 702 673 1748 430 832 332 675 3 776 809 344 1190 762 446 274 1076 2458 18 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7025 8067 894 1115 701 1378 * * 9(D) 469 1542 1605 42 1969 1375 320 1627 36 1747 919 219 1080 65 522 1153 479 670 2048 1444 19 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7025 8067 894 1115 701 1378 * * 10(S) 586 1385 736 829 2829 1309 1317 2735 1241 2859 2047 1835 1910 1051 1602 2833 881 1997 2969 2410 20 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7025 8067 894 1115 701 1378 * * 11(P) 526 1890 212 1174 2213 1420 120 1928 1548 1905 1024 80 1551 317 111 415 763 1521 2093 1450 21 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7025 8067 894 1115 701 1378 * * 12(D) 1339 2192 1728 1307 2485 1401 248 2248 996 2199 1317 28 1622 180 495 529 648 1804 2382 1658 22 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7025 8067 894 1115 701 1378 * * 13(Y) 984 983 1421 1092 837 1586 819 1015 973 1266 608 1029 1341 855 1218 770 660 790 1317 3268 23 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7025 8067 894 1115 701 1378 * * 14(L) 877 775 2589 1991 1160 2382 1170 1029 960 1849 533 1790 2366 1386 1652 1454 810 77 1124 783 24 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7025 8067 894 1115 701 1378 * * 15(N) 730 2232 123 1772 2530 1420 275 2293 34 2239 1364 1865 1127 1325 496 569 693 1850 2418 1697 25 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7025 8067 894 1115 701 1378 * * 16(S) 406 1619 309 191 1903 406 102 1593 986 317 789 1064 1504 317 181 1301 358 1237 1903 1286 26 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7025 8067 894 1115 701 1378 * * 17(M) 675 566 2528 1928 364 2184 2126 218 1592 743 2772 1654 2210 1313 1560 1256 614 1711 990 635 27 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 18(D) 749 1575 2786 299 2062 1566 768 1153 635 1769 1097 535 1902 450 1112 799 731 1070 2328 1751 28 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 19(K) 1232 1674 736 295 2360 1534 473 2014 2326 2058 1228 486 1165 74 221 634 677 1596 2264 1735 29 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 20(Y) 2075 2643 1686 985 266 2358 869 2644 1801 2467 2120 1177 2732 1384 2229 1931 2114 2488 905 4141 30 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 21(W) 2473 2122 3795 3534 136 3469 1296 944 2887 1176 449 2889 3469 2568 2708 2895 2429 1317 5419 316 31 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 22(R) 1335 1772 2246 1513 2127 2222 955 1242 35 1675 1197 1424 2426 659 3303 1507 1331 1211 2220 1868 32 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 23(A) 1948 1167 891 1114 1539 1605 710 688 551 1278 608 703 1861 451 928 724 609 1295 1870 1386 33 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 24(A) 2275 1339 993 558 2142 1451 682 1733 1352 1909 1113 663 1785 334 564 587 928 1336 2220 1732 34 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 25(N) 1216 1775 1072 1034 1473 1967 1153 1429 1009 767 1165 3461 2339 1043 1259 1335 1316 1331 1982 1273 35 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 26(Y) 2131 1808 3471 3249 538 3299 814 1516 2817 864 742 2495 3305 2356 2649 2520 2091 784 205 4093 36 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 27(L) 2321 1872 4652 4075 539 4266 3092 2178 3803 2345 2067 3915 3747 2978 3487 3494 2215 175 2130 2157 37 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 28(S) 482 1136 1858 1915 3072 1315 1996 2905 2045 3106 2272 1475 1651 1840 2205 3016 873 2011 3242 2908 38 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 29(V) 845 1090 3817 3373 1522 3377 2794 2147 3161 649 421 3056 3362 2955 3177 2598 1419 2690 2606 2197 39 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 30(G) 1283 2067 837 965 3177 3042 1462 3119 1171 3141 2429 1111 2331 2005 1448 1320 1499 2538 3099 2665 40 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 31(Q) 974 2142 780 270 2589 1791 328 2217 1581 2142 1329 487 1903 3083 262 876 671 1860 2250 1764 41 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 32(L) 1375 1127 3503 2949 697 3071 2054 1887 2629 2035 342 2656 2990 2251 2534 2214 1157 428 1759 1511 42 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 33(Y) 1937 2273 1926 1563 232 2608 650 2164 1538 2035 1599 1501 2740 960 541 1929 1851 2048 794 4044 43 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 34(L) 2871 2457 4231 4103 1033 3803 3165 541 3734 3130 31 3935 3797 3286 3484 3713 2869 1136 2394 2220 44 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 35(K) 565 1769 542 1150 2042 1544 133 1707 1529 200 1362 220 1622 291 1492 490 487 1367 1948 1377 45 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 36(D) 733 2199 2095 230 2550 256 327 2314 1307 2269 1390 100 1681 97 574 1114 708 1861 2454 1735 46 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 37(N) 1190 2662 1381 127 3153 1520 768 2990 757 2945 2156 3138 1961 405 1416 1061 1253 2479 3137 2313 47 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 38(P) 1777 2801 1868 561 3633 1918 1524 3634 1721 3620 2978 881 3543 1257 2350 1648 1944 3091 3510 2972 48 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 39(C) 1617 2940 3806 3389 788 2963 2369 4 2967 2637 197 2927 3094 2569 2799 2284 1672 278 1950 1680 49 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 40(M) 1138 1232 1092 1546 625 2468 1290 56 1453 1974 2682 1610 2481 1228 1655 1558 1079 283 1540 1223 50 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 41(H) 30 439 11 363 394 739 699 414 463 757 501 286 577 639 165 23 111 312 697 25 51 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 42(K) 784 2190 977 302 2645 1847 256 2259 2452 2136 1309 504 1900 1339 1312 874 863 1897 2202 1742 52 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 43(E) 648 1915 516 1778 2222 1578 162 1899 439 163 1023 238 1667 1297 1557 552 568 1536 2057 1482 53 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 44(P) 1091 2157 710 354 2472 1828 2409 2236 1296 2195 1418 568 2737 72 88 1007 1021 1903 2268 1751 54 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 45(L) 1149 1110 2516 1945 1939 2543 1114 128 1051 1953 309 1805 2527 1346 1480 1629 1078 306 1080 509 55 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 46(T) 695 1464 884 358 1467 1703 361 1272 1547 1430 690 543 1822 57 149 739 2166 1045 1654 1676 56 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 47(R) 1626 2138 1332 620 2693 1985 403 2249 1356 2167 1392 747 2067 11 2526 1100 1051 1929 2243 1870 57 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 48(E) 378 1941 703 1824 2252 1386 100 2000 289 1956 1046 24 1516 349 1172 476 427 1561 2134 1447 58 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 49(D) 923 2559 2894 160 2929 1519 635 2712 524 2688 1882 189 1895 1449 1131 904 1111 2258 2898 2120 59 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 50(V) 1091 1040 3230 3029 1839 2191 2521 453 2807 1107 816 2387 2674 2598 2801 1506 1141 3085 2726 2349 60 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 51(K) 1016 1925 1252 838 2825 1834 758 2369 3094 2408 1667 917 2126 380 14 1094 1125 1973 2524 2145 61 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 52(H) 477 400 2258 1664 1050 1948 2313 1043 1367 239 404 1388 365 1083 1355 1008 417 915 715 1744 62 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 53(H) 1023 1918 1234 509 2208 1943 3152 947 1315 1817 1055 662 1981 64 1825 993 895 1542 1997 1584 63 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 54(P) 1113 913 1858 1496 1358 1604 1238 756 1296 958 503 1306 2592 1174 1479 817 686 602 1858 1474 64 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 55(I) 846 1093 3818 3374 1512 3374 2792 2798 3161 633 411 3056 3360 2951 3174 2596 1422 2077 2600 2195 65 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 56(G) 2594 2690 3304 3623 4328 3747 3462 4761 3953 4671 4212 3320 3352 3748 3779 2839 2981 4004 3668 4222 66 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 57(H) 3205 3079 2723 2890 2110 3046 5295 4135 2617 3813 3561 2886 3482 2833 2620 3291 3356 3895 2397 1681 67 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 58(W) 4114 3274 4214 4453 1722 3496 2681 4109 4149 3619 3615 4044 3885 4032 3816 4355 4248 4133 6191 1329 68 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 59(G) 2594 2690 3304 3623 4328 3747 3462 4761 3953 4671 4212 3320 3352 3748 3779 2839 2981 4004 3668 4222 69 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 60(T) 1213 1674 2755 2906 3163 1922 2659 2698 2788 3105 2612 2311 2600 2708 2753 1463 3819 2197 3286 3156 70 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 61(C) 859 2735 3125 2633 1043 2353 1765 2619 2314 561 137 2173 2560 2046 2258 1540 1442 680 1746 1384 71 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 62(P) 1490 891 2212 1956 2077 1371 1689 1274 1796 1861 1203 1479 2954 1611 1936 653 684 816 2478 2131 72 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 63(G) 382 1055 1768 1819 3131 2891 1951 2904 2042 3099 2224 1384 1043 1771 2236 1022 775 1958 3301 2980 73 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 64(Q) 1532 1831 1845 1487 1032 2520 1248 707 794 1863 184 1573 2625 2991 924 1768 1488 990 1859 1397 74 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 65(N) 1040 1491 1193 1126 1577 1932 1248 1438 1138 1433 1011 3384 2307 1120 1396 1256 1150 579 2101 1473 75 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 66(F) 2629 2152 4261 4009 3717 3972 1470 438 3655 1364 94 3253 3712 2829 3311 3250 2543 971 709 261 76 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 67(I) 1794 1354 4306 3856 1267 3968 3273 2808 3666 1262 89 3607 3721 3300 3605 3222 1758 1880 2695 2428 77 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 68(Y) 1692 1412 3458 3020 258 3136 1081 1186 2630 646 31 2459 3064 2167 2476 2272 1624 329 544 3829 78 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 69(A) 2378 934 2147 1885 2779 826 1775 2485 1781 2703 1842 1381 1860 1577 1989 1353 1040 1689 2983 2654 79 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 70(H) 3205 3079 2723 2890 2110 3046 5295 4135 2617 3813 3561 2886 3482 2833 2620 3291 3356 3895 2397 1681 80 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 71(L) 2379 1915 4669 4151 621 4360 3248 1934 3862 2632 553 4017 3845 3099 3585 3641 2289 24 2250 2225 81 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 72(N) 2171 2655 1458 1748 3334 2364 2267 3943 2365 3936 3437 4205 2932 2205 2608 2224 2439 3392 3253 2909 82 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 73(R) 1929 2343 2336 1600 825 2607 677 2317 45 2153 1653 1506 2681 679 3382 1922 1792 2176 1232 2267 83 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 74(V) 503 701 3099 2506 1199 2516 1425 486 2168 1370 379 2113 2515 1800 2039 1621 848 2021 1247 916 84 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 75(I) 2091 1746 3971 3840 1676 3532 3289 3684 3581 659 693 3562 3674 3445 3521 3194 2146 449 2877 2493 85 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 76(K) 1051 1939 439 69 2445 1557 349 2132 2202 2116 1268 1786 1753 70 141 646 704 1719 2284 1701 86 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 77(K) 1384 2800 1892 69 3174 1733 742 2966 2973 2873 2105 400 2087 365 775 1179 1372 2535 2949 2288 87 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 78(Y) 2118 2084 2592 2218 1983 3049 2937 1851 1966 1662 1339 1862 3049 1795 2085 2125 2036 1809 122 3287 88 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 79(N) 1107 2701 1326 223 2992 1724 586 2799 472 2733 1910 2292 1876 1179 1098 894 1109 2331 2923 2116 89 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 80(L) 1690 1806 2247 1903 837 2782 1536 464 1204 2489 81 1939 2835 1986 1289 2052 1642 850 1841 1451 90 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 81(D) 1292 2956 2598 202 3213 1558 693 3045 851 2958 2170 2502 1967 318 1262 1043 1304 2569 3134 2291 91 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 82(M) 866 1094 2548 2220 1176 1993 1696 374 1810 470 4232 1867 2385 1729 1855 1260 1587 400 1960 1618 92 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 83(F) 1725 1400 3934 3394 2962 3436 2055 325 3076 1369 437 3005 3243 2505 2863 2595 1653 1644 1488 995 93 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 84(Y) 2852 2251 3961 3935 2281 3735 476 168 3538 1379 1299 2630 3641 2627 3134 2925 2756 1840 233 4154 94 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 85(I) 1759 1305 4328 3965 1743 4047 3735 3086 3833 588 522 3726 3872 3681 3903 3364 1752 2354 3250 2813 95 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 86(M) 700 1108 1144 2046 1106 1894 710 177 572 648 2421 857 1997 497 908 934 657 1171 1602 1155 96 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 87(G) 2594 2690 3304 3623 4328 3747 3462 4761 3953 4671 4212 3320 3352 3748 3779 2839 2981 4004 3668 4222 97 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 88(P) 526 1169 1848 1957 3095 1343 2052 2946 2123 3152 2333 1513 3407 1917 2261 1632 922 2053 3267 2938 98 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 89(G) 2594 2690 3304 3623 4328 3747 3462 4761 3953 4671 4212 3320 3352 3748 3779 2839 2981 4004 3668 4222 99 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 90(H) 3205 3079 2723 2890 2110 3046 5295 4135 2617 3813 3561 2886 3482 2833 2620 3291 3356 3895 2397 1681 100 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 91(G) 2594 2690 3304 3623 4328 3747 3462 4761 3953 4671 4212 3320 3352 3748 3779 2839 2981 4004 3668 4222 101 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 92(H) 30 439 11 363 394 739 699 414 463 757 501 286 577 639 165 23 111 312 697 25 102 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 93(G) 1432 1081 2340 2519 3309 3113 2408 3041 2687 3327 2482 1740 2062 2353 2689 702 903 2058 3476 3301 103 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 94(Q) 1326 2187 825 789 2878 1884 1131 2784 631 2752 2063 977 2646 3194 881 1313 1428 2357 2787 2310 104 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 95(V) 1470 815 2475 2234 1874 1135 1780 892 2064 1657 1016 1624 2020 1814 2121 717 687 2477 2329 1999 105 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 96(M) 883 899 2454 1951 595 991 1285 31 1626 1077 3599 1754 2362 1420 1669 1376 878 131 1359 1024 106 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 97(V) 1205 914 3493 2955 1023 3025 2085 1649 2672 917 32 2617 2982 2375 2606 2169 1373 2267 1903 1552 107 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 98(S) 1719 1145 1296 1130 2738 1260 1357 2461 1234 2624 1769 1171 1844 1070 1576 2383 686 1744 2874 2423 108 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 99(N) 809 2086 98 33 2593 1494 488 2361 147 2346 1503 2681 1787 1970 607 984 825 1908 2521 1851 109 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 100(S) 878 890 1818 1458 1679 1418 1269 1166 1293 665 837 1224 1904 1156 1509 2469 610 869 2068 1669 110 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 101(Y 3588 2709 4120 4323 1034 3994 419 2613 3834 2031 2055 2741 3905 2826 3349 3280 3472 2736 3673 4312 111 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 102(L) 2496 2027 4721 4202 560 4432 3262 1210 3887 2852 611 4095 3876 3086 3587 3729 2397 232 2211 2201 112 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 103(D 1791 3663 3414 1659 3853 1647 1043 3807 1343 3679 3054 251 2201 725 2227 1438 1872 3282 3836 2831 113 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 104(G 2594 2690 3304 3623 4328 3747 3462 4761 3953 4671 4212 3320 3352 3748 3779 2839 2981 4004 3668 4222 114 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 105(S) 342 976 2177 2128 2921 1229 2008 2606 2073 2888 2058 1512 1936 1871 2189 2669 2327 1779 3151 2842 115 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 106(Y 2652 2206 3673 3556 726 3580 692 1359 3066 982 936 2620 3538 2500 2832 2837 2587 1595 24 4185 116 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 107(T) 502 1110 2201 2282 3095 1340 2204 2779 2341 3075 2268 1660 2066 2111 2412 729 3346 1945 3276 3043 117 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 108(E) 1163 2811 1893 2460 3072 1535 594 2884 1337 2797 1978 140 1898 203 1072 933 1159 2412 2972 2157 118 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 109(I) 561 338 2506 1896 1485 1980 829 1917 1557 155 470 1524 2033 1236 1465 1055 777 309 776 1058 119 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 110(Y 3611 2705 4166 4409 2531 4042 397 2533 3989 1938 1983 2747 3929 2851 3444 3294 3491 2684 345 4289 120 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 111(P) 2931 2878 3420 3706 4181 2925 3468 4621 3859 4490 4165 3491 4225 3781 3695 3182 3279 4087 3594 4064 121 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 112(E) 785 2281 2 1960 2596 1496 297 2348 660 2275 1399 1934 1705 1721 381 626 738 1906 2432 1737 122 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 113(I) 1722 1438 3424 3080 131 3150 1169 2917 2681 503 344 2513 3144 2305 2554 2347 1683 39 645 2880 123 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 114(T) 1213 1674 2755 2906 3163 1922 2659 2698 2788 3105 2612 2311 2600 2708 2753 1463 3819 2197 3286 3156 124 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 115(Q 1609 2594 1354 790 3182 2213 519 2737 2527 2514 1787 903 2297 3031 525 1483 1437 2432 2460 2150 125 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 116(D 2784 3432 4016 1200 4140 2466 2197 4505 2621 4365 3956 1551 3014 2039 3232 2593 2938 4046 3710 3552 126 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 117(E) 495 1766 320 2020 2044 1449 133 1708 1298 1759 898 124 1562 295 155 410 632 108 1998 1374 127 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 118(E) 1485 2051 202 1709 2406 1511 270 2112 1557 2089 1228 183 1688 156 234 586 657 1706 2272 1633 128 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 119(G 2594 2690 3304 3623 4328 3747 3462 4761 3953 4671 4212 3320 3352 3748 3779 2839 2981 4004 3668 4222 129 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 120(M 2568 2113 4714 4154 462 4418 3155 99 3799 2469 3538 4062 3833 2972 3479 3691 2450 587 2108 2139 130 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 121(Q 447 1907 448 115 2176 1605 2296 1980 143 2000 1194 367 1799 3046 194 715 752 1626 2150 1543 131 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 122(K 1448 2495 1440 709 3095 2161 406 2616 2874 2399 1636 1544 2197 22 1483 1326 1270 2295 2364 2043 132 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 123(L) 2430 1987 4659 4071 2109 4260 2828 67 3779 2463 2382 3869 3717 2880 3418 3473 2306 623 1874 1724 133 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 124(F) 2144 1762 4034 3706 3638 3656 1587 1655 3381 102 23 3073 3507 2734 3134 2896 2093 241 906 16 134 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 125(K 1297 2351 1405 643 2938 2076 377 2482 2603 2306 1527 768 2120 51 1973 899 1147 2149 2307 1957 135 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 126(Q 753 1991 692 91 2345 1670 1700 2009 519 1957 1111 334 1745 2315 1728 656 1061 1645 2095 1558 136 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 127(F) 3342 2776 4026 4232 4354 3545 1431 2315 4038 1801 1900 3299 3780 3350 3645 3490 3420 2566 739 349 137 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 128(S) 897 1462 2333 2543 3185 1640 2474 3294 2686 3497 2780 1973 2360 2483 2703 3465 1316 2413 3310 3025 138 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 129(F) 1908 1643 3261 2963 3263 3050 509 1219 2636 1197 851 2199 3071 2085 2476 2189 1297 1211 77 2811 139 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 130(P) 540 1180 1850 1974 3103 1352 2072 2962 2149 3170 2355 1527 3509 1943 2279 1382 938 2068 3276 2949 140 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 131(G 1432 1081 2340 2519 3309 3113 2408 3041 2687 3327 2482 1740 2062 2353 2689 702 903 2058 3476 3301 141 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 132(G 2594 2690 3304 3623 4328 3747 3462 4761 3953 4671 4212 3320 3352 3748 3779 2839 2981 4004 3668 4222 142 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 133(V 1322 992 3697 3181 1158 3230 2353 2069 2914 914 126 2840 3160 2624 2849 2396 641 2352 2122 1765 143 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 134(P) 2498 1088 2250 2290 3023 1338 2177 2668 2306 2970 2190 1656 2760 2080 2382 710 882 1878 3231 2984 144 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 135(S) 897 1462 2333 2543 3185 1640 2474 3294 2686 3497 2780 1973 2360 2483 2703 3465 1316 2413 3310 3025 145 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 136(H 3205 3079 2723 2890 2110 3046 5295 4135 2617 3813 3561 2886 3482 2833 2620 3291 3356 3895 2397 1681 146 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 137(A 1947 745 2823 2285 1846 2348 1328 1572 1988 154 207 1942 2439 1677 1944 1473 836 359 1231 781 147 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 138(A 2029 2389 1954 92 2936 1492 703 2716 623 2715 1906 1712 1898 331 1237 881 1089 2229 2931 2168 148 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 139(P) 1935 1116 2232 2301 3058 1358 2206 2705 2336 3009 2238 1674 3272 2114 2406 739 914 1913 3260 3019 149 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 140(E) 1349 2706 162 2827 3109 1785 624 2820 1799 2704 1931 437 2073 230 334 1160 1298 2421 2774 2189 150 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 141(T) 645 1000 2671 2474 1925 1699 2057 385 2200 1416 1020 1885 2274 2057 2247 1011 3256 1160 2570 2205 151 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 142(P) 2931 2878 3420 3706 4181 2925 3468 4621 3859 4490 4165 3491 4225 3781 3695 3182 3279 4087 3594 4064 152 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 143(H 30 439 11 363 394 739 699 414 463 757 501 286 577 639 165 23 111 312 697 25 153 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 144(G 2594 2690 3304 3623 4328 3747 3462 4761 3953 4671 4212 3320 3352 3748 3779 2839 2981 4004 3668 4222 154 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 145(S) 1426 949 2380 2419 3107 1201 2226 2819 2411 3105 2246 1618 1944 2131 2456 3026 742 1874 3336 3080 155 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 146(I) 2091 1746 3971 3840 1676 3532 3289 3684 3581 659 693 3562 3674 3445 3521 3194 2146 449 2877 2493 156 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 147(H 3205 3079 2723 2890 2110 3046 5295 4135 2617 3813 3561 2886 3482 2833 2620 3291 3356 3895 2397 1681 157 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 148(E) 2641 3308 896 3732 3966 2458 2043 4105 2128 4016 3555 1531 2959 1842 2560 2479 2750 3722 3563 3385 158 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 149(G 2594 2690 3304 3623 4328 3747 3462 4761 3953 4671 4212 3320 3352 3748 3779 2839 2981 4004 3668 4222 159 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 150(G 2594 2690 3304 3623 4328 3747 3462 4761 3953 4671 4212 3320 3352 3748 3779 2839 2981 4004 3668 4222 160 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 151(E) 669 2323 22 3133 3044 1600 954 2706 853 2824 2095 473 2059 614 1376 1068 1279 2254 3056 2376 161 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 152(L) 2871 2457 4231 4103 1033 3803 3165 541 3734 3130 31 3935 3797 3286 3484 3713 2869 1136 2394 2220 162 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 153(G 2594 2690 3304 3623 4328 3747 3462 4761 3953 4671 4212 3320 3352 3748 3779 2839 2981 4004 3668 4222 163 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 154(Y 3482 2868 3701 3919 238 3552 1112 3000 3638 2516 2526 3027 3772 3101 3341 3418 3527 3071 441 4711 164 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 155(S) 2313 937 2414 2419 3070 1197 2205 2774 2385 3059 2200 1614 1936 2106 2436 2518 728 1846 3302 3046 165 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 156(L) 2364 1901 4660 4144 630 4349 3246 2015 3856 2597 544 4006 3841 3101 3583 3629 2275 2 2255 2227 166 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 157(S) 643 1153 1942 1769 1726 1574 1553 1424 1560 381 1167 1475 2124 1504 1711 2968 911 1168 2234 1683 167 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 158(H 1208 1519 1818 1600 796 2137 4436 1042 1134 1422 996 1514 2459 1277 1260 1442 1296 1290 1367 527 168 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 159(G 1625 934 2312 2287 3120 2515 2142 2866 2304 3096 2206 1553 1910 2010 2401 1405 705 1884 3324 3069 169 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 160(Y 553 516 2670 2093 1648 2153 831 125 1753 275 301 1673 2204 1407 1649 1238 604 1878 648 2121 170 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 161(G 2594 2690 3304 3623 4328 3747 3462 4761 3953 4671 4212 3320 3352 3748 3779 2839 2981 4004 3668 4222 171 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 162(A 3438 1472 2846 3040 3287 1726 2735 2840 3028 3257 2662 2236 2447 2798 2944 1216 1387 2183 3405 3320 172 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 163(I) 1759 1304 4329 3967 1748 4050 3739 3054 3836 593 525 3728 3874 3686 3907 3367 1751 2401 3255 2817 173 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 164(M 2389 1957 4609 4018 1345 4211 2821 78 3724 2123 3673 3822 3688 2852 3378 3418 2267 598 1888 1767 174 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 165(D 2784 3432 4016 1200 4140 2466 2197 4505 2621 4365 3956 1551 3014 2039 3232 2593 2938 4046 3710 3552 175 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 166(N 2171 2655 1458 1748 3334 2364 2267 3943 2365 3936 3437 4205 2932 2205 2608 2224 2439 3392 3253 2909 176 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 167(P) 2931 2878 3420 3706 4181 2925 3468 4621 3859 4490 4165 3491 4225 3781 3695 3182 3279 4087 3594 4064 177 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 168(D 1220 2896 2608 1191 3149 1530 651 2978 602 2895 2089 1257 1925 269 1273 1237 1229 2498 3083 2233 178 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 169(L) 1037 975 2376 275 735 2518 1412 367 1669 2017 219 1835 2540 1484 1766 1625 988 1640 1541 1207 179 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 170(I) 1918 1539 4076 3666 2083 3676 2142 3117 3379 145 94 3221 3504 2843 3198 2911 1872 334 1508 773 180 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 171(V 2267 970 2988 2782 1867 1873 2265 74 2556 1277 903 2107 2419 2332 2560 1182 982 2421 2604 2249 181 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 172(A 2401 935 2092 1865 2596 1235 1739 2211 1758 2497 1687 1386 1817 1571 1946 528 1549 1545 2846 2512 182 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 173(C 152 3737 3346 3008 1521 2075 2187 405 2700 983 575 2269 2516 2413 2614 1349 959 2580 2260 1908 183 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 174(V 1189 1030 2667 1142 1294 2870 1904 1998 2055 599 276 2175 2880 1914 2228 1998 1157 2462 2134 1721 184 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 175(V 1753 1297 4327 3965 1769 4049 3747 2530 3837 621 545 3725 3875 3695 3913 3366 1745 2911 3274 2827 185 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 176(G 2594 2690 3304 3623 4328 3747 3462 4761 3953 4671 4212 3320 3352 3748 3779 2839 2981 4004 3668 4222 186 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 177(D 2784 3432 4016 1200 4140 2466 2197 4505 2621 4365 3956 1551 3014 2039 3232 2593 2938 4046 3710 3552 187 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 178(G 2594 2690 3304 3623 4328 3747 3462 4761 3953 4671 4212 3320 3352 3748 3779 2839 2981 4004 3668 4222 188 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 179(E) 2641 3308 896 3732 3966 2458 2043 4105 2128 4016 3555 1531 2959 1842 2560 2479 2750 3722 3563 3385 189 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 180(A 3124 934 2490 2562 3082 1203 2297 2766 2535 3081 2234 1670 1954 2236 2534 920 746 1844 3332 3091 190 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 181(E) 2641 3308 896 3732 3966 2458 2043 4105 2128 4016 3555 1531 2959 1842 2560 2479 2750 3722 3563 3385 191 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 182(T) 1213 1674 2755 2906 3163 1922 2659 2698 2788 3105 2612 2311 2600 2708 2753 1463 3819 2197 3286 3156 192 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 183(G 1080 1108 2311 2509 3336 3223 2424 3074 2706 3361 2521 1751 2082 2372 2708 729 932 2089 3493 3324 193 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 184(P) 2931 2878 3420 3706 4181 2925 3468 4621 3859 4490 4165 3491 4225 3781 3695 3182 3279 4087 3594 4064 194 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 185(L) 1369 1384 3092 2737 825 2649 2073 44 2334 2571 187 2437 2847 2125 2306 1915 1278 290 1941 1678 195 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 186(A 2912 1000 2422 2157 1636 1634 1753 816 1876 1128 1988 1694 2169 1741 1956 923 861 675 2256 1899 196 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 187(T) 973 953 2483 2484 2838 1268 2195 2209 2352 2704 1971 1675 1991 2131 2373 617 3427 1554 3148 2888 197 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 188(S) 437 1083 2006 2144 3262 1583 2197 3097 2380 3307 2434 1562 2008 2077 2485 2965 857 2074 3426 3160 198 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 189(W 4114 3274 4214 4453 1722 3496 2681 4109 4149 3619 3615 4044 3885 4032 3816 4355 4248 4133 6191 1329 199 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 190(H 1323 1832 1402 994 2149 2268 3183 1595 684 1541 1023 1062 2358 2478 951 1335 1257 1461 476 569 200 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 191(S) 1326 873 2274 2002 2263 1287 1735 1644 1844 2124 1388 1463 1915 1637 1990 2623 646 880 2605 2269 201 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 192(N 573 1630 864 62 1860 1478 310 843 39 1634 837 2440 1658 65 526 537 638 1154 1980 1380 202 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 193(K 1324 1987 1657 1059 2437 2134 750 1568 3217 1940 1374 1107 2304 391 150 1379 1277 548 2320 1952 203 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 194(H 30 439 11 363 394 739 699 414 463 757 501 286 577 639 165 23 111 312 697 25 204 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 195(F) 2721 2160 3990 3906 3360 3716 572 1433 3510 1177 922 2674 3597 2614 3108 2899 2622 1671 137 2722 205 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 196(I) 1769 1321 4308 3881 1430 3998 3403 2941 3713 744 240 3637 3769 3419 3696 3267 1740 2073 2864 2543 206 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 197(N 1660 3287 1909 69 3610 1646 1046 3616 1275 3528 2867 3539 2186 731 2069 1373 1757 3086 3624 2697 207 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 198(P) 2931 2878 3420 3706 4181 2925 3468 4621 3859 4490 4165 3491 4225 3781 3695 3182 3279 4087 3594 4064 208 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 199(R 780 954 1042 463 1021 1681 388 1232 838 838 132 612 1756 173 1322 665 418 574 1339 893 209 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 200(N 622 2053 110 1285 2366 1450 199 2102 1009 2062 1176 2124 1614 238 276 491 1247 1677 2241 1566 210 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 201(D 1369 2674 3374 58 3453 1571 1095 3380 1289 3368 2659 394 2124 784 2016 1149 1529 2785 3505 2662 211 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 202(G 2594 2690 3304 3623 4328 3747 3462 4761 3953 4671 4212 3320 3352 3748 3779 2839 2981 4004 3668 4222 212 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 203(A 2246 790 2416 1988 1270 1821 1490 1726 1755 905 386 1651 2194 1557 1854 1027 949 146 1839 1474 213 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 204(V 1771 1603 3750 3689 2037 3050 3231 403 3479 1154 1076 3246 3399 3383 3437 2628 1917 3536 3074 2677 214 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 205(L) 2871 2457 4231 4103 1033 3803 3165 541 3734 3130 31 3935 3797 3286 3484 3713 2869 1136 2394 2220 215 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 206(P) 2931 2878 3420 3706 4181 2925 3468 4621 3859 4490 4165 3491 4225 3781 3695 3182 3279 4087 3594 4064 216 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 207(I) 2091 1746 3971 3840 1676 3532 3289 3684 3581 659 693 3562 3674 3445 3521 3194 2146 449 2877 2493 217 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 208(L) 2475 2006 4716 4195 570 4423 3262 1353 3887 2819 603 4084 3871 3089 3590 3715 2377 194 2218 2208 218 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 209(H 1047 2367 130 12 2443 1653 3422 2425 11 2366 1567 2352 1902 1348 351 897 1003 2040 2389 1693 219 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 210(L) 1968 1597 4244 3772 812 3841 2955 582 3473 2391 322 3529 3600 2960 3315 3097 1926 2070 2269 2100 220 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 211(N 2171 2655 1458 1748 3334 2364 2267 3943 2365 3936 3437 4205 2932 2205 2608 2224 2439 3392 3253 2909 221 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 212(G 2594 2690 3304 3623 4328 3747 3462 4761 3953 4671 4212 3320 3352 3748 3779 2839 2981 4004 3668 4222 222 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 213(F) 3581 2688 4165 4401 3865 4034 401 2491 3981 1903 1944 2746 3921 2847 3438 3284 3462 2647 341 3054 223 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 214(K 2620 2961 2461 2046 3743 2791 1570 3603 3784 3387 2839 2048 3039 1260 465 2604 2536 3331 3001 2988 224 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 215(I) 2091 1746 3971 3840 1676 3532 3289 3684 3581 659 693 3562 3674 3445 3521 3194 2146 449 2877 2493 225 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 216(A 2478 935 2423 2429 3066 1197 2208 2769 2392 3055 2196 1617 1936 2112 2440 2351 728 1843 3300 3045 226 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 217(N 1558 3098 310 1592 3431 1657 945 3340 959 3271 2572 3495 2146 611 1578 1301 1621 2860 3383 2547 227 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 218(P) 1664 2231 2030 1695 3192 2240 1444 3029 556 2975 2319 1668 3549 1149 1833 1745 1774 2613 2848 2650 228 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 219(T) 1213 1674 2755 2906 3163 1922 2659 2698 2788 3105 2612 2311 2600 2708 2753 1463 3819 2197 3286 3156 229 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 220(I) 1764 1326 4294 3932 1658 3987 3644 3354 3776 498 454 3684 3833 3599 3826 3303 1760 1799 3153 2738 230 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 221(F) 2604 2127 4552 4135 2542 4236 2203 115 3820 2542 500 3692 3785 2927 3457 3508 2493 769 1337 643 231 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 222(S) 846 947 2243 2217 3137 1905 2112 2888 2254 3112 2218 1528 1909 1968 2369 2545 708 1899 3331 3067 232 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 223(R 1454 3144 2763 2070 2728 2193 1414 2383 460 2518 1907 1827 2567 1160 3510 1626 1582 2068 2596 2366 233 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 224(M 434 912 967 653 960 1640 368 1179 682 787 2592 569 1718 161 575 617 633 341 1294 841 234 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 225(S) 1102 2199 1764 235 3206 1518 1117 3089 1203 3129 2368 513 2060 804 1816 2841 1292 2470 3271 2541 235 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 226(D 1724 3448 3411 103 3726 1642 1058 3734 1342 3628 2991 1884 2196 745 2191 1408 1822 3197 3743 2768 236 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 227(E) 391 2060 303 2460 2453 1565 312 2145 213 2119 1272 258 1745 110 1394 657 720 1747 2290 1683 237 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 228(E) 843 2213 113 2617 2562 1553 357 2268 1012 2237 1392 212 1769 60 324 706 776 1863 2412 1764 238 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 229(L) 2324 1864 4634 4121 657 4319 3239 2224 3838 2493 517 3975 3830 3106 3578 3596 2239 73 2272 2231 239 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 230(H 505 1236 775 228 1373 1593 2318 963 872 1178 425 432 1693 14 290 578 1743 825 1578 1083 240 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 231(C 561 2259 1449 1476 2294 1417 164 2034 1472 2003 1111 57 1572 275 300 433 510 1609 2194 1510 241 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 232(Y 2882 2267 4018 3995 2580 3778 511 1658 3593 923 1152 2675 3663 2654 3169 2969 2779 1865 205 3751 242 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 233(F) 3342 2776 4026 4232 4354 3545 1431 2315 4038 1801 1900 3299 3780 3350 3645 3490 3420 2566 739 349 243 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 234(H 559 1906 319 1741 2211 1478 2491 1923 355 1899 1021 132 1591 303 791 452 1019 1527 2083 1454 244 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 235(G 456 1098 1993 2173 3296 2968 2244 3141 2461 3355 2485 1583 2025 2139 2549 1723 881 2104 3461 3203 245 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 236(M 1532 1301 3405 2864 57 3050 1438 56 2477 1096 3967 2484 2939 2042 2342 2169 1455 370 989 2079 246 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 237(G 1038 1765 926 1191 3372 3233 1826 3379 1926 3481 2725 1285 2271 1628 2264 1145 1377 2554 3401 2980 247 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 238(W 3553 2695 4100 4288 1022 3972 428 2592 3792 2021 2038 2737 3890 2815 3323 3265 3440 2712 4944 3677 248 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 239(H 693 2112 279 1837 2456 1534 2434 2176 1108 2098 1217 167 1666 1691 39 560 624 1753 2235 1604 249 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 240(P) 853 1252 2460 2393 2139 1784 2109 974 2210 1748 1364 1924 3453 2108 2271 1167 1152 1448 2678 2307 250 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 812 7108 1241 894 1115 701 1378 * * 241(Y 906 1365 1255 688 4 1806 48 1150 314 1157 604 702 1897 107 2106 965 822 1011 491 2916 251 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 27 6323 7365 894 1115 292 2448 * * 242(Y 476 775 1198 1077 755 1738 505 1586 528 626 43 777 1822 410 817 751 649 143 1165 1945 252 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 243(F) 1857 1592 3755 3460 3638 3255 1454 16 3144 214 206 2800 3287 2604 2951 2505 1863 1480 853 99 253 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 244(V 1203 1434 1278 1166 1756 2236 1499 75 1412 1192 819 1390 2525 1332 1696 1574 1248 2962 2384 1907 254 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 245(H 30 439 11 363 394 739 699 414 463 757 501 286 577 639 165 23 111 312 697 25 255 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 246(E) 1028 2230 1464 1545 2560 1444 343 2324 77 2285 1411 93 1690 78 635 1220 730 1876 2478 1750 256 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 247(G 802 892 1664 1291 1774 2148 1181 1363 1177 617 906 1109 1839 1027 1436 836 560 1000 2098 1700 257 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 248(F) 737 1710 1482 1223 2205 1628 301 1513 202 1603 865 339 1790 72 681 695 688 1259 1410 1271 258 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 249(D 1662 3241 3494 12 3550 1708 991 3449 907 3360 2675 360 2197 658 1631 1384 1715 2975 3451 2636 259 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 250(K 374 1824 783 927 2121 1368 40 525 1057 1830 920 824 249 411 141 290 342 1433 2020 1349 260 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 251(E) 424 2247 1450 1906 2557 209 314 2325 37 2276 1395 75 659 111 595 584 710 1874 2461 1729 261 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 570 7108 1648 894 1115 701 1378 * * 252(D 1129 2752 2889 513 3030 1216 532 2959 669 2896 2188 2229 1714 198 1413 860 1196 2465 3041 2140 262 149 500 233 43 379 399 105 627 212 466 721 275 394 45 96 359 117 370 295 248 754 2619 2035 2949 200 992 1008 * * 253(H 696 961 1261 873 178 1785 3652 1381 427 516 92 889 1934 521 602 945 673 203 756 22 273 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 25 6457 7499 894 1115 1666 546 * * 254(M 1561 1309 3310 2786 99 3163 1984 569 2364 1604 3948 2656 2936 1983 2284 2361 1493 116 1472 1272 274 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 25 6457 7499 894 1115 777 1263 * * 255(K 475 1891 137 1054 2221 1359 30 1946 1632 1890 999 2 1483 1488 30 1135 411 1523 2049 1403 275 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 19 6850 7893 894 1115 1215 813 * * 256(V 1531 1105 4020 3576 1227 3695 3034 2323 3387 793 62 3320 3500 3101 3368 2941 1501 2608 2576 2243 276 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 19 6850 7893 894 1115 1215 813 * * 257(H 1244 2186 246 319 1501 1694 4400 2492 417 2436 1795 1396 2075 501 711 1152 1294 2140 1842 960 277 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 19 6850 7893 894 1115 1215 813 * * 258(R 675 1969 242 1673 2340 1479 164 2032 420 1985 1141 153 1471 261 1760 560 613 1643 2136 1550 278 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 19 6850 7893 894 1115 453 1893 * * 259(E) 377 1835 1113 1421 2128 1382 69 1857 329 1840 939 29 1493 377 1329 322 374 19 2039 1371 279 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 260(M 2315 1899 4526 3941 2102 4105 2681 70 3638 1954 1509 3704 3629 2798 3304 3301 2198 582 1802 1604 280 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 261(A 2818 1372 1321 1214 2015 1557 2623 2096 1108 2284 1581 1154 2074 1121 1340 881 964 1649 2321 1711 281 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 262(K 793 2177 220 1469 2548 1561 289 2263 2325 2196 1334 214 1738 141 150 898 737 1844 2341 1711 282 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 263(A 1931 776 2058 1612 1165 1647 1206 433 1382 945 310 1370 2014 1216 1530 832 1640 293 1672 1305 283 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 264(M 1519 1247 3637 3077 1725 3081 1493 37 2718 1089 3901 2600 2950 2153 2487 2201 1437 379 1806 356 284 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 265(D 1378 3094 2801 1763 3349 1556 771 3202 821 3114 2349 164 2006 409 1544 1050 1411 2709 3307 2411 285 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 266(W 426 1591 384 832 1812 1435 111 1488 966 1565 720 131 1532 300 184 757 1423 1162 2539 1242 286 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 267(V 1401 623 2516 1981 808 2086 1255 1461 1718 518 36 1694 2253 1472 1752 293 658 1939 1375 1008 287 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 268(W 857 644 3104 2515 1895 2435 1142 1168 2142 39 357 2021 2440 1737 1961 1534 797 2161 2512 335 288 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 269(E) 1213 2845 1928 2618 3115 1534 660 2926 607 2864 2064 150 1929 281 1267 980 833 2457 3061 2225 289 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 270(E) 1357 2719 146 2868 3119 1782 634 2833 1719 2718 1948 434 2077 242 361 1166 1309 2433 2790 2201 290 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 271(I) 2091 1746 3971 3840 1676 3532 3289 3684 3581 659 693 3562 3674 3445 3521 3194 2146 449 2877 2493 291 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 272(C 1095 2832 1254 506 2502 1969 1775 2085 2238 2024 1241 660 2004 96 1706 1025 959 1790 2126 1723 292 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 273(D 1682 2499 2383 115 3011 1503 717 2803 660 2789 1985 1727 1918 347 1292 922 1145 2313 2999 2214 293 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 274(I) 2091 1746 3971 3840 1676 3532 3289 3684 3581 659 693 3562 3674 3445 3521 3194 2146 449 2877 2493 294 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 275(Q 1627 2584 1823 913 3248 2312 430 2707 1993 2449 1710 969 2306 3139 1660 1504 1407 2413 2377 2126 295 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 276(K 949 2020 181 1024 2347 1460 176 2077 2067 2031 1143 996 1606 264 187 477 547 1653 2203 1543 296 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 277(H 1280 1815 274 261 2114 1392 1493 1840 1076 1818 916 946 1489 1230 102 316 245 1427 2010 1354 297 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 278(A 3438 1472 2846 3040 3287 1726 2735 2840 3028 3257 2662 2236 2447 2798 2944 1216 1387 2183 3405 3320 298 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 279(R 836 1922 846 238 2255 1752 261 1878 494 1879 1072 484 1828 1594 2601 770 741 369 2053 1564 299 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 280(E) 531 1917 152 1776 2240 1412 166 1971 212 1955 1067 76 1565 1351 287 854 1223 1553 2155 1485 300 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 281(N 756 2277 1639 264 2584 735 320 2357 45 2301 1420 1764 1681 1399 604 499 725 1902 2482 1746 301 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 1243 7108 811 894 1115 701 1378 * * 282(T) 391 254 749 541 1519 523 582 1080 464 1400 646 345 1090 368 718 1584 1914 580 1775 1351 302 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 36 5902 6944 894 1115 2014 410 * * 283(N 1419 678 16 71 1606 696 281 1227 86 1488 735 2022 1167 9 450 17 79 791 1790 1251 303 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 36 5902 6944 894 1115 655 1455 * * 284(N 690 2252 1313 1338 2538 1300 208 2319 1491 2250 1381 1988 1564 211 512 509 660 1861 2424 1672 304 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 20 6788 7830 894 1115 421 1984 * * 285(D 1304 3019 2680 1285 3255 1544 705 3097 703 3006 2219 1388 1348 331 1399 1044 1321 2611 3195 2320 305 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 286(H 30 439 11 363 394 739 699 414 463 757 501 286 577 639 165 23 111 312 697 25 306 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 287(M 1003 593 321 1362 615 2007 849 1868 1149 373 2071 1286 2064 949 1296 1049 514 769 1128 747 307 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 288(M 426 945 995 468 1082 1580 1667 681 299 938 2081 605 1724 229 641 445 2054 498 1403 940 308 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 289(R 1619 2365 2075 1064 2835 2363 469 2305 1823 763 1488 1073 2345 67 2817 1563 1402 2096 2243 1997 309 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 290(P) 2931 2878 3420 3706 4181 2925 3468 4621 3859 4490 4165 3491 4225 3781 3695 3182 3279 4087 3594 4064 310 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 291(Q 802 740 2393 1814 1799 2272 1035 123 1529 790 401 1638 2287 2034 1573 1335 740 1456 1046 619 311 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 292(W 3500 2678 4063 4226 995 3934 449 2563 3717 2009 2016 2732 3866 2798 3276 3242 3394 2681 5405 3043 312 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 293(P) 2931 2878 3420 3706 4181 2925 3468 4621 3859 4490 4165 3491 4225 3781 3695 3182 3279 4087 3594 4064 313 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 294(M 2406 2296 3638 3594 1525 3105 2824 1047 3121 596 5043 3293 3425 3046 2996 2911 2552 1398 2513 2207 314 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 295(I) 2034 1595 4395 3950 974 4099 3249 3261 3681 1202 174 3748 3784 3212 3556 3388 1988 627 2497 2285 315 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 296(I) 1757 1299 4333 3971 1766 4057 3755 2831 3844 614 541 3733 3880 3700 3920 3375 1749 2665 3275 2830 316 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 297(F) 2590 2117 4458 4084 3096 4144 1957 191 3757 2203 405 3544 3752 2887 3401 3410 2485 812 1135 337 317 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 298(R 2957 3022 3318 2735 3796 2998 1968 3912 846 3631 3157 2611 3280 1724 4056 3026 2913 3650 3096 3185 318 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 299(T) 2050 905 2305 2099 2720 1206 1895 2371 1974 2648 1815 1479 1888 1753 2118 1016 2546 1624 2968 2665 319 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 300(P) 2931 2878 3420 3706 4181 2925 3468 4621 3859 4490 4165 3491 4225 3781 3695 3182 3279 4087 3594 4064 320 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 301(K 2620 2961 2461 2046 3743 2791 1570 3603 3784 3387 2839 2048 3039 1260 465 2604 2536 3331 3001 2988 321 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 302(G 2594 2690 3304 3623 4328 3747 3462 4761 3953 4671 4212 3320 3352 3748 3779 2839 2981 4004 3668 4222 322 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 303(W 4114 3274 4214 4453 1722 3496 2681 4109 4149 3619 3615 4044 3885 4032 3816 4355 4248 4133 6191 1329 323 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 304(T) 353 974 2215 2202 2896 1238 2053 2560 2141 2867 2052 1547 1951 1940 2229 1380 3291 1759 3143 2840 324 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 305(C 845 3595 3279 3420 3342 3140 2815 3103 3232 3426 2671 2300 2379 2934 3057 1107 1278 2270 3401 3393 325 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 306(P) 2931 2878 3420 3706 4181 2925 3468 4621 3859 4490 4165 3491 4225 3781 3695 3182 3279 4087 3594 4064 326 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 307(K 2620 2961 2461 2046 3743 2791 1570 3603 3784 3387 2839 2048 3039 1260 465 2604 2536 3331 3001 2988 327 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 308(M 402 813 1034 898 582 1639 381 420 330 698 1526 611 1715 218 668 337 1310 272 1195 1104 328 149 500 234 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 176 3166 8150 310 2369 701 1378 * * 309(W 1600 1315 3759 3181 295 3243 1874 1788 2803 1756 538 2802 3074 2285 2600 2379 1521 139 4106 936 330 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 310(D 1718 3250 3403 27 3066 1736 2667 3486 1211 3381 2732 400 2240 752 1917 1439 1784 3023 3204 2299 331 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 311(G 1708 2667 1499 644 3784 3218 1636 3799 1910 3782 3127 941 2466 1384 2554 1618 1929 3151 3633 3136 332 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 312(K 708 2195 956 221 2514 1474 242 2264 2240 2194 1308 1084 1658 1697 314 557 656 1821 2352 1662 333 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 313(P) 762 1504 940 454 1424 1749 441 1317 1946 1439 747 629 1988 176 300 813 723 1105 1603 967 334 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 314(I) 1232 1121 3291 2953 1431 2753 2412 3122 2652 599 404 2590 2982 2506 2674 2020 1485 773 2421 2034 335 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 315(E) 2641 3308 896 3732 3966 2458 2043 4105 2128 4016 3555 1531 2959 1842 2560 2479 2750 3722 3563 3385 336 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 316(G) 1046 1820 755 1003 3341 2933 1683 3325 1756 3410 2651 2259 2235 1456 2142 1125 1359 2537 3369 2897 337 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 317(S) 897 1462 2333 2543 3185 1640 2474 3294 2686 3497 2780 1973 2360 2483 2703 3465 1316 2413 3310 3025 338 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 318(W 3425 2618 4155 4330 3859 3949 484 2303 3894 1728 1759 2779 3865 2847 3389 3247 3323 2490 3919 1426 339 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 319(R 1940 2623 2163 1317 2718 2498 2633 2821 557 2576 1920 1294 2549 305 3406 1859 1734 2575 2279 1885 340 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 320(A 2865 932 2456 2481 3066 1198 2239 2763 2443 3058 2203 1637 1941 2156 2474 1737 732 1840 3307 3058 341 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 321(H 3205 3079 2723 2890 2110 3046 5295 4135 2617 3813 3561 2886 3482 2833 2620 3291 3356 3895 2397 1681 342 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 322(Q 2562 2904 1886 1971 3251 2661 2079 3690 1565 3469 3081 2107 3091 4371 1665 2585 2674 3411 3077 2821 343 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 323(I) 1757 1300 4333 3971 1764 4057 3754 2871 3843 611 539 3733 3880 3699 3919 3375 1749 2625 3273 2829 344 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 324(P) 2931 2878 3420 3706 4181 2925 3468 4621 3859 4490 4165 3491 4225 3781 3695 3182 3279 4087 3594 4064 345 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 325(I) 1899 1466 4358 3871 1002 3993 3149 2479 3652 1948 170 3634 3690 3153 3516 3233 1847 1460 2466 2295 346 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 326(P) 1615 1136 2222 2310 3082 1373 2227 2731 2358 3036 2272 1689 3445 2139 2424 761 938 1937 3279 3042 347 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 327(V 776 978 1248 821 1609 1418 840 1138 733 1453 710 1478 1781 614 1074 1249 508 1777 1905 1460 348 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 328(D 1973 1793 2321 222 2801 1401 864 2543 758 2614 1785 457 1858 510 1294 684 896 1978 2851 2198 349 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 329(Q 1008 2231 986 310 2701 1861 258 2315 1642 2173 1344 511 1911 2494 1827 403 883 1946 2224 1767 350 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 330(D 1037 2659 2253 1117 2928 1504 509 2731 1324 2651 1807 1958 1830 106 940 826 1023 2261 2827 2032 351 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 331(H 935 2308 2421 146 2574 1508 2726 2388 331 2395 1577 187 1822 135 890 787 1407 1976 2588 1852 352 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 332(M 650 1996 1349 1466 2242 1448 264 1958 34 1982 2809 101 597 150 493 533 608 1580 2227 1552 353 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 333(E) 1263 2196 74 2930 2933 1571 907 2588 778 2712 1962 469 2014 562 1290 991 1179 2137 2954 2290 354 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 334(H 1082 2079 440 1291 845 1821 3880 1961 281 1944 1264 509 2017 267 666 989 1031 1699 1298 1603 355 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 335(F) 1005 431 2207 1610 1664 1966 799 1077 332 809 427 1386 2016 1061 1337 1023 432 229 879 519 356 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 336(H 30 439 11 363 394 739 699 414 463 757 501 286 577 639 165 23 111 312 697 25 357 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 337(D 996 2602 2234 1640 2872 1493 484 2671 308 2598 1748 104 783 1779 913 792 980 2205 2778 1990 358 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 338(A 1659 679 1813 1259 745 1962 855 1055 1039 483 101 1222 2054 1021 1227 1014 543 1478 1248 857 359 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 339(L) 2871 2457 4231 4103 1033 3803 3165 541 3734 3130 31 3935 3797 3286 3484 3713 2869 1136 2394 2220 360 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 340(V 891 1677 1034 464 1912 1849 400 1350 1652 1575 859 635 1938 1706 124 916 803 2029 1916 1472 361 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 341(D 869 2445 2202 1077 2729 120 389 2516 1310 2445 1574 1245 1739 29 730 684 841 2053 2620 1858 362 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 342(W 4114 3274 4214 4453 1722 3496 2681 4109 4149 3619 3615 4044 3885 4032 3816 4355 4248 4133 6191 1329 363 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 343(L) 2642 2168 4788 4235 465 4513 3241 96 3878 2891 2122 4166 3888 3023 3544 3807 2521 607 2140 2183 364 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 344(E) 588 2030 204 1763 2360 1459 147 2094 1522 2032 1135 95 1592 1372 129 847 526 1661 2190 1529 365 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 345(S) 897 1462 2333 2543 3185 1640 2474 3294 2686 3497 2780 1973 2360 2483 2703 3465 1316 2413 3310 3025 366 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 346(Y 3482 2868 3701 3919 238 3552 1112 3000 3638 2516 2526 3027 3772 3101 3341 3418 3527 3071 441 4711 367 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 347(K 1779 2672 1588 963 3316 2329 561 2833 3294 2578 1879 1030 2399 1424 589 1653 1579 2550 2489 2237 368 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 348(P) 2931 2878 3420 3706 4181 2925 3468 4621 3859 4490 4165 3491 4225 3781 3695 3182 3279 4087 3594 4064 369 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 349(E) 1283 2980 2293 2465 3222 1551 680 3054 846 2961 2167 151 1956 302 1278 1029 1292 2574 3140 2287 370 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 350(E) 2641 3308 896 3732 3966 2458 2043 4105 2128 4016 3555 1531 2959 1842 2560 2479 2750 3722 3563 3385 371 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 351(L) 2871 2457 4231 4103 1033 3803 3165 541 3734 3130 31 3935 3797 3286 3484 3713 2869 1136 2394 2220 372 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 352(F) 3342 2776 4026 4232 4354 3545 1431 2315 4038 1801 1900 3299 3780 3350 3645 3490 3420 2566 739 349 373 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 353(D 1170 2662 2768 152 3076 1523 716 2882 677 2851 2058 1883 1940 347 1326 973 1251 2402 3056 2245 374 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 354(E) 1031 2152 91 2416 2507 1517 346 2220 982 2201 1350 188 1736 71 388 657 752 1810 2390 1731 375 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 355(N 1247 2802 2354 164 3217 1531 760 3060 767 2997 2213 2573 1971 395 1447 1055 1298 2553 3188 2339 376 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 356(G 2594 2690 3304 3623 4328 3747 3462 4761 3953 4671 4212 3320 3352 3748 3779 2839 2981 4004 3668 4222 377 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 357(H 969 1896 1173 279 2199 1387 1573 1938 276 1912 1011 34 1518 1339 236 939 421 1513 2105 1426 378 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 358(V 1097 861 3273 2702 715 2790 1749 1319 2390 1604 246 2365 510 2047 2299 1909 1047 2041 1564 1253 379 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 359(K 627 934 1396 826 1049 1906 680 1291 2041 745 142 930 1983 512 793 921 762 912 1475 1045 380 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 360(D 417 2242 2268 951 2534 1439 289 2301 2 2249 1365 64 475 1392 551 566 688 1853 2431 1702 381 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 361(D 756 2213 2042 1784 2484 1455 327 2233 59 287 1351 90 1688 90 612 609 722 1813 2423 1705 382 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 362(I) 1758 1302 4331 3969 1754 4053 3745 3003 3839 600 530 3730 3876 3691 3912 3370 1751 2469 3262 2822 383 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 363(K 489 1736 399 869 1996 1468 108 1679 1463 680 849 140 1560 1326 65 410 492 1326 1942 1336 384 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 364(E) 1607 1738 268 2001 2459 1405 567 2171 308 2231 1384 339 1737 179 802 1027 695 1699 2473 1839 385 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 365(I) 521 410 2443 1845 1682 2040 909 1831 1536 217 389 581 2091 1245 1495 1112 673 1143 913 551 386 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 366(M 1028 781 3313 2728 655 2725 1667 2105 2396 791 2636 2332 2694 2027 2260 1840 662 1307 1459 1146 387 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 367(P) 2931 2878 3420 3706 4181 2925 3468 4621 3859 4490 4165 3491 4225 3781 3695 3182 3279 4087 3594 4064 388 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 368(K 420 1711 319 889 1985 1406 81 1683 1799 1711 835 80 1506 351 148 802 593 66 1944 1311 389 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 369(G 1210 2695 1728 114 3097 2436 713 2899 1005 2848 2055 236 1965 340 1178 1009 1238 2429 3026 2250 390 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 370(D 1173 2847 2185 2156 3091 1523 1843 2917 527 2832 2014 1280 1899 221 1180 937 1174 2440 3014 2176 391 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 371(K 733 1639 900 275 1870 1727 272 1476 1449 1052 793 476 1798 105 1100 1022 645 1222 1849 1373 392 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 372(R 2124 2788 2710 1449 3624 2637 576 2974 1929 2640 1977 1344 2596 161 3417 2010 1819 2745 2479 2379 393 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 373(M 2186 1770 4484 3908 547 4083 2924 1892 3619 1562 3822 3715 3642 2876 3343 3290 2088 106 2072 2052 394 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 374(A 2525 1036 2401 2529 3239 2375 2366 2958 2629 3244 2393 1719 2028 2303 2639 657 852 1993 3425 3237 395 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 375(M 1980 959 1279 892 1385 1524 842 905 763 1211 2928 1065 1852 673 1053 675 559 683 1743 1304 396 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 376(N 1426 2353 948 735 2920 1991 796 2769 68 2666 1942 3441 2268 432 1742 1361 1415 2383 2592 2147 397 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 377(P) 566 1202 1850 1997 3117 1371 2101 2989 2185 3198 2391 1549 3625 1982 2305 1013 967 2094 3288 2963 398 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 276 7108 2582 894 1115 701 1378 * * 378(I) 820 759 2281 1810 939 2345 1389 2404 1595 425 18 1689 2430 1447 1742 1465 804 1594 1641 1225 399 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 19 6850 7893 894 1115 1215 813 * * 379(T) 1785 732 2107 1994 2428 1065 1767 1811 1860 2282 1548 1351 1750 1665 1956 390 2954 1225 2740 2439 400 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 19 6850 7893 894 1115 453 1893 * * 380(N 1558 3098 310 1592 3431 1657 945 3340 959 3271 2572 3495 2146 611 1578 1301 1621 2860 3383 2547 401 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 381(G 2030 1051 2376 2525 3268 2820 2382 2992 2652 3277 2428 1726 2039 2322 2659 672 869 2018 3447 3263 402 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 382(G 1587 1953 2017 2080 898 3122 1483 2565 2178 2576 2101 1908 2727 2015 2307 1743 1817 2251 1457 2463 403 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 195 7108 3069 894 1115 701 1378 * * 383(K 585 1070 1140 510 1174 1742 358 751 1799 916 239 641 1802 103 873 754 507 1342 1431 1005 404 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 18 6931 7973 894 1115 505 1760 * * 384(I) 732 1273 1962 516 1501 1754 770 1970 673 1163 572 718 1976 522 1082 902 1162 263 1908 1404 405 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 385(H 542 1652 558 18 1880 1542 2080 339 360 1605 778 241 1147 1217 1461 492 468 1229 1864 1303 406 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 386(K 682 1971 580 740 2315 1607 158 1989 2284 162 1077 529 1686 280 734 578 595 1611 2087 1523 407 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 387(H 30 439 11 363 394 739 699 414 463 757 501 286 577 639 165 23 111 312 697 25 408 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 388(P) 415 2181 1668 189 2543 1458 400 2293 146 2281 1425 138 2240 1274 694 643 769 1860 2492 1780 409 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 389(L) 1646 1694 2904 2680 1050 2701 2138 547 2336 2673 123 2479 1513 2198 2336 2107 1735 860 2077 1777 410 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 390(D 430 1571 1536 153 1777 1435 129 307 1131 1533 700 889 1538 275 255 380 371 598 1838 1235 411 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 391(L) 2871 2457 4231 4103 1033 3803 3165 541 3734 3130 31 3935 3797 3286 3484 3713 2869 1136 2394 2220 412 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 392(P) 2931 2878 3420 3706 4181 2925 3468 4621 3859 4490 4165 3491 4225 3781 3695 3182 3279 4087 3594 4064 413 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 393(D 123 3021 2652 1083 3262 1545 715 3102 722 3016 2232 2288 1970 344 1422 1053 1333 2618 3208 2331 414 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 394(W 1183 940 3255 2753 1534 2632 772 1086 2365 1132 71 2086 2628 1856 2132 1737 1115 336 3856 1849 415 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 395(R 1025 2540 1775 1142 2847 1561 472 2610 136 2532 1695 183 1846 64 2638 837 993 2170 2686 1970 416 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 396(D 1284 2963 2433 216 3207 1549 693 3050 653 2963 2174 2058 1961 2341 1315 1034 1299 2571 3142 2286 417 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 397(Y 3621 2707 4176 4424 2970 4050 394 2539 4003 1942 1987 2749 3933 2855 3451 3299 3499 2690 349 4083 418 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 398(C 1443 2403 1117 1144 997 1629 520 480 452 234 131 706 1773 348 779 656 418 324 1362 922 419 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 399(V 1742 1294 4287 3868 1507 3983 3425 2151 3706 586 316 3621 3770 3449 3709 3254 1716 2871 2925 2571 420 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 400(D 730 2244 2028 1115 2543 1450 283 2307 1800 2248 1364 74 1665 147 498 571 502 1859 2424 1702 421 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 401(N 410 1793 248 958 2083 143 54 1808 1270 1797 898 1557 1482 391 146 306 586 53 2000 1340 422 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 402(Q 430 1511 419 58 1825 563 219 1494 116 1597 764 1033 1570 1217 353 1025 404 892 1898 1308 423 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 403(K 42 2025 1238 279 2338 1408 138 2090 1948 2034 1127 1026 1554 1361 253 410 491 1645 2204 1512 424 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 404(E) 625 2031 279 1765 2367 1499 161 2087 353 2026 1140 139 1309 1668 922 500 559 1667 2180 1544 425 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 1300 7108 769 894 1115 701 1378 * * 405(Y 1580 1211 2445 2254 2497 2497 449 742 1944 476 322 1430 2471 1343 1796 1668 1507 820 1088 3543 426 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 38 5845 6888 894 1115 560 1636 * * 406(G 1371 2394 1717 285 3391 3003 1248 3345 1454 3349 2666 589 2148 975 2092 1270 1563 2745 3295 2730 427 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 19 6850 7893 894 1115 1215 813 * * 407(H 1177 1467 267 124 1912 1286 2549 1618 132 1695 852 1230 1499 193 333 681 372 1236 1957 1349 428 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 19 6850 7893 894 1115 1215 813 * * 408(N 352 1085 738 561 1975 1933 822 1555 672 1841 1078 2120 1685 547 1041 473 532 758 2198 1695 429 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 725 6850 1371 894 1115 1215 813 * * 409(W 643 1210 455 1427 532 1453 184 970 26 1073 478 390 1652 90 302 705 607 812 4240 180 430 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 348 6155 2320 894 1115 1881 457 * * 410(G 461 838 976 1119 2120 3148 1219 2101 1326 2273 1700 986 1505 1212 1430 653 781 1558 1923 1928 431 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 38 5845 6888 894 1115 560 1636 * * 411(Q 389 1056 645 768 1124 1508 222 612 34 219 187 343 1598 1815 346 489 329 1199 1410 918 432 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 19 6850 7893 894 1115 453 1893 * * 412(F) 523 749 1182 612 1262 1690 436 327 1202 257 75 713 1761 315 572 681 361 558 1146 721 433 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 413(Q 838 2270 27 1546 2609 1503 383 2361 43 2318 1463 158 1752 2803 531 937 810 1929 2497 1800 434 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 414(D 2081 2191 2846 271 3192 1547 1150 2913 1247 3052 2319 548 2085 840 1860 1066 1314 2364 3266 2572 435 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 415(M 746 729 1725 1241 1091 1556 930 594 1043 922 3667 1111 404 891 1250 698 914 410 1509 1123 436 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 416(I) 556 796 1468 861 813 1848 559 1435 1067 271 42 912 1905 460 1342 865 488 747 1213 814 437 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 417(E) 738 1306 528 1243 1448 1471 165 1079 130 90 452 231 1559 192 330 328 268 267 1616 1069 438 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 418(W 1686 1388 3865 3280 313 3346 1998 1605 2910 2183 589 2918 3142 2351 2689 2486 1602 216 3506 1080 439 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 419(G 497 1132 1973 2195 3328 3237 2289 3194 2532 3410 2549 1611 2057 2201 2603 897 926 2151 3487 3237 440 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 420(K 632 1777 1362 250 2087 1381 86 1807 1431 1808 914 51 1497 355 193 817 369 425 2019 1362 441 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 421(Y 3621 2707 4176 4424 2953 4049 394 2539 4002 1942 1987 2749 3933 2854 3451 3299 3499 2690 349 4093 442 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 422(C 642 1771 2908 2287 1063 2214 1093 1477 1913 1497 494 1833 2239 1538 1745 1304 765 369 942 612 443 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 423(R 1005 2443 1533 92 2784 1584 463 2543 57 2471 1639 1177 1853 56 2770 836 972 2113 2612 1931 444 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 424(D 1148 2724 2898 174 3006 1564 566 2796 951 2711 1895 178 1903 1815 851 933 1132 2344 2872 2108 445 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 425(V 1646 1230 4148 3696 1391 3782 3103 2535 3497 323 1927 3416 3609 3214 3466 3018 1616 2552 2681 2340 446 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 426(I) 401 900 3283 2756 1640 2752 1735 2834 2451 121 132 2375 2775 2110 2360 1897 1085 615 1519 1079 447 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 427(K 834 1941 615 845 2257 1704 312 1778 2586 1890 1101 402 1823 95 65 764 756 788 2124 1599 448 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 428(N 857 1859 733 254 2082 1745 340 1726 1897 679 1022 2134 1861 45 77 813 778 1455 2024 1510 449 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 429(N 2171 2655 1458 1748 3334 2364 2267 3943 2365 3936 3437 4205 2932 2205 2608 2224 2439 3392 3253 2909 450 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 430(P) 1140 1553 2272 2156 1935 1979 1912 1526 1824 1503 1339 1897 3724 1871 1917 1397 1391 1428 2485 2072 451 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 431(H 464 1881 1234 255 2190 1407 2368 1921 1008 1884 981 50 1513 375 482 349 1263 1498 2063 1403 452 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 432(N 761 2121 1175 150 2581 1441 443 2337 205 2327 1467 2649 1731 36 757 420 1163 1879 2535 1825 453 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 433(F) 1857 1592 3755 3460 3638 3255 1454 16 3144 214 206 2800 3287 2604 2951 2505 1863 1480 853 99 454 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 434(R 2957 3022 3318 2735 3796 2998 1968 3912 846 3631 3157 2611 3280 1724 4056 3026 2913 3650 3096 3185 455 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 435(L) 886 722 2913 2339 608 1022 1423 1356 2029 1682 294 2025 2497 1718 1972 1579 842 1130 1337 1008 456 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 436(F) 3342 2776 4026 4232 4354 3545 1431 2315 4038 1801 1900 3299 3780 3350 3645 3490 3420 2566 739 349 457 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 437(G 468 1108 1987 2183 3307 3093 2261 3159 2489 3374 2507 1593 2035 2163 2570 1410 895 2119 3471 3217 458 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 438(H 30 439 11 363 394 739 699 414 463 757 501 286 577 639 165 23 111 312 697 25 459 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 439(P) 2931 2878 3420 3706 4181 2925 3468 4621 3859 4490 4165 3491 4225 3781 3695 3182 3279 4087 3594 4064 460 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 440(D 2784 3432 4016 1200 4140 2466 2197 4505 2621 4365 3956 1551 3014 2039 3232 2593 2938 4046 3710 3552 461 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 441(E) 2641 3308 896 3732 3966 2458 2043 4105 2128 4016 3555 1531 2959 1842 2560 2479 2750 3722 3563 3385 462 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 442(T) 439 840 1804 1403 1361 1523 1133 816 1185 681 536 1214 1931 1069 1378 642 2718 603 1788 1389 463 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 443(A 1657 1594 404 1150 1818 1465 154 1471 969 1566 1309 173 1572 248 202 422 412 1160 1861 1272 464 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 444(S) 897 1462 2333 2543 3185 1640 2474 3294 2686 3497 2780 1973 2360 2483 2703 3465 1316 2413 3310 3025 465 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 445(N 2171 2655 1458 1748 3334 2364 2267 3943 2365 3936 3437 4205 2932 2205 2608 2224 2439 3392 3253 2909 466 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 446(R 1905 2715 2334 1200 3487 2506 485 2858 1904 2546 1846 1167 2464 1439 3148 1780 1627 2602 2423 2263 467 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 447(L) 2639 2158 4642 4194 1586 4340 2490 51 3890 2827 573 3860 3832 2979 3524 3629 2524 732 1565 1006 468 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 448(Q 549 1725 324 98 1793 1481 198 1668 163 1723 892 839 1618 2328 290 843 499 1331 1882 1794 469 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 449(A 2109 2374 1307 1077 2738 326 514 2505 344 2485 1647 151 1804 118 934 766 932 2058 2698 1950 470 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 450(A 1280 376 2219 1624 1011 1905 779 1073 1336 235 411 1368 1975 1065 1339 339 580 955 851 492 471 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 451(F) 2619 2089 3969 3846 3011 3673 612 1304 3453 1530 788 2671 3552 2584 3066 2851 2522 1556 92 2879 472 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 452(E) 400 2342 1163 2306 2664 1461 401 2438 168 2392 1527 102 1735 14 743 947 818 1984 2583 1837 473 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 453(H 424 834 1231 741 1039 183 2171 588 582 902 199 797 1793 479 878 660 1442 1664 1404 972 474 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 454(T) 825 1543 1050 931 2411 1607 1105 2069 661 2229 1555 978 2065 1333 881 930 3234 1676 2544 2072 475 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 455(N 948 2217 192 59 2753 871 516 2480 1579 2431 1604 2747 1874 115 403 831 947 2039 2574 1954 476 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 456(R 2118 2789 2710 1441 3626 2635 569 2972 2088 2636 1971 1337 2591 153 3343 2003 1811 2743 2476 2375 477 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 276 7108 2582 894 1115 701 1378 * * 457(Q 2077 2533 1425 1467 2808 2285 1598 3134 1059 2958 2537 1616 2694 4251 1184 2087 2168 2871 2703 2369 478 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 19 6850 7893 894 1115 1215 813 * * 458(W 3599 2878 3805 3989 1220 3156 2211 3491 3618 3057 3027 3568 3532 3525 3338 3829 3720 3529 6135 830 479 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 19 6850 7893 894 1115 1215 813 * * 459(M 718 1070 1476 726 732 1903 713 152 681 774 3189 882 1991 541 1003 968 669 221 1408 983 480 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 19 6850 7893 894 1115 1215 813 * * 460(E) 598 1592 123 1874 1828 1337 104 1490 228 43 755 967 1490 296 254 355 375 1164 1876 1258 481 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 19 6850 7893 894 1115 1215 813 * * 461(P) 556 1744 1230 129 2482 1292 454 2234 252 2255 1403 147 1758 66 790 781 633 1727 2476 1801 482 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 1043 6850 983 894 1115 1215 813 * * 462(Y 1345 1235 1689 1605 1266 1964 117 925 1303 775 592 1236 2169 1119 1309 1450 1367 939 590 3938 483 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 38 5845 6888 894 1115 560 1636 * * 463(I) 1578 1151 4046 3597 1151 3733 3046 3001 3402 771 18 3355 3517 3085 3372 2981 1545 1808 2540 2237 484 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 19 6850 7893 894 1115 453 1893 * * 464(H 620 1972 273 140 2293 1493 2910 2020 1123 1988 1114 1414 1632 244 127 1037 563 1615 2160 1523 485 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 465(E) 290 1861 861 1446 981 1390 101 1871 273 1861 968 953 303 339 239 355 411 1463 2069 1398 486 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 466(P) 546 1977 862 1025 2262 1409 155 2002 204 546 1086 48 1748 1636 313 420 495 1582 2175 1489 487 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 467(W 477 661 1479 920 458 1795 520 227 729 529 143 1513 1868 571 958 304 420 1257 2766 1177 488 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 468(D 2784 3432 4016 1200 4140 2466 2197 4505 2621 4365 3956 1551 3014 2039 3232 2593 2938 4046 3710 3552 489 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 469(Q 1336 2814 130 2632 3112 1670 702 2921 403 2830 2067 313 2037 2655 853 1120 1328 2494 2954 2238 490 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 470(W 1164 1540 661 1080 2483 2257 1994 1245 947 1304 742 745 2310 770 1263 1253 1097 1115 2555 1863 491 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 471(M 459 840 1110 504 907 1674 501 397 456 727 3234 715 1788 352 771 770 419 680 1288 853 492 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 472(H 1702 1846 317 119 2155 1480 1717 1860 1008 1870 1009 1392 1616 236 157 482 520 1482 2080 1459 493 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 473(P) 1735 849 2213 1898 1800 1446 1575 902 1724 1568 933 1474 2335 1533 1863 706 666 1535 2237 1882 494 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 474(K 373 1864 703 990 2179 1360 29 1928 1276 1877 955 4 1460 1260 124 542 486 1483 2049 1367 495 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 475(G 2594 2690 3304 3623 4328 3747 3462 4761 3953 4671 4212 3320 3352 3748 3779 2839 2981 4004 3668 4222 496 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 476(R 1165 2238 1063 503 2794 820 418 2401 526 2281 1492 670 2047 2040 2681 1075 1064 2048 2328 1916 497 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 477(V 823 660 2789 2196 638 2422 1317 1842 1836 191 1866 1913 2436 1596 1039 1512 768 2028 1296 949 498 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 478(V 1748 1297 4301 3885 1515 3999 3451 2579 3726 585 321 3638 3782 3470 3730 3273 1722 2581 2943 2589 499 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 479(D 1067 2475 2632 1644 2683 1544 584 1013 469 2422 1654 180 1882 209 1071 894 1059 1933 2725 1977 500 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 340 7108 2304 894 1115 701 1378 * * 480(S) 717 1527 107 1500 2213 1229 377 1911 112 1988 1149 164 1554 7 597 1919 503 1461 2242 1620 501 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 20 6788 7830 894 1115 421 1984 * * 481(Q 790 1076 1563 1026 923 2037 834 178 669 369 1897 1109 2120 2919 906 1087 746 1186 1510 1104 502 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 482(L) 2871 2457 4231 4103 1033 3803 3165 541 3734 3130 31 3935 3797 3286 3484 3713 2869 1136 2394 2220 503 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 483(S) 897 1462 2333 2543 3185 1640 2474 3294 2686 3497 2780 1973 2360 2483 2703 3465 1316 2413 3310 3025 504 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 484(E) 2641 3308 896 3732 3966 2458 2043 4105 2128 4016 3555 1531 2959 1842 2560 2479 2750 3722 3563 3385 505 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 485(H 3205 3079 2723 2890 2110 3046 5295 4135 2617 3813 3561 2886 3482 2833 2620 3291 3356 3895 2397 1681 506 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 486(Q 1064 1426 1517 1022 242 2170 535 884 583 715 396 1085 2235 3051 804 1225 994 864 835 1693 507 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 487(C 1096 3020 1354 606 1109 1571 566 613 497 939 1724 707 1762 380 838 622 428 433 1460 1015 508 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 488(E) 1401 2897 169 3007 3185 1677 753 3009 498 2916 2170 319 2069 1880 966 1173 1402 2580 3040 2307 509 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 489(H 30 439 11 363 394 739 699 414 463 757 501 286 577 639 165 23 111 312 697 25 510 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 490(G 2594 2690 3304 3623 4328 3747 3462 4761 3953 4671 4212 3320 3352 3748 3779 2839 2981 4004 3668 4222 511 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 491(W 2172 1773 3771 3493 3388 3396 717 939 3099 528 2179 2555 3307 2387 2796 2550 2089 1165 3733 1010 512 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 492(L) 2620 2150 4778 4218 461 4493 3219 96 3870 2765 2758 4143 3872 3009 3534 3777 2498 607 2129 2177 513 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 493(E) 1401 2897 169 3007 3185 1677 753 3009 498 2916 2170 319 2069 1880 966 1173 1402 2580 3040 2307 514 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 494(G 2172 1046 2384 2525 3259 2719 2377 2982 2644 3267 2417 1723 2035 2315 2652 666 864 2010 3440 3255 515 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 495(Y 3482 2868 3701 3919 238 3552 1112 3000 3638 2516 2526 3027 3772 3101 3341 3418 3527 3071 441 4711 516 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 496(T) 971 915 2809 2296 857 2442 1626 433 1998 1151 92 2043 2569 1769 2033 1592 2524 1200 1666 1332 517 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 497(L) 2871 2457 4231 4103 1033 3803 3165 541 3734 3130 31 3935 3797 3286 3484 3713 2869 1136 2394 2220 518 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 498(T) 1213 1674 2755 2906 3163 1922 2659 2698 2788 3105 2612 2311 2600 2708 2753 1463 3819 2197 3286 3156 519 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 499(G 2594 2690 3304 3623 4328 3747 3462 4761 3953 4671 4212 3320 3352 3748 3779 2839 2981 4004 3668 4222 520 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 500(R 2957 3022 3318 2735 3796 2998 1968 3912 846 3631 3157 2611 3280 1724 4056 3026 2913 3650 3096 3185 521 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 501(H 999 1647 1206 1105 1780 1755 4476 2154 805 2276 1630 1129 2202 979 999 1106 1428 1786 2106 1395 522 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 502(G 2594 2690 3304 3623 4328 3747 3462 4761 3953 4671 4212 3320 3352 3748 3779 2839 2981 4004 3668 4222 523 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 503(I) 1567 1223 3950 3399 2055 3419 2370 2527 3104 944 309 3048 3245 2620 2927 2582 1507 1309 1899 1632 524 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 504(W 3425 2618 4155 4330 3859 3949 484 2303 3894 1728 1759 2779 3865 2847 3389 3247 3323 2490 3919 1426 525 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 505(A 2014 983 1788 1434 2527 1231 1430 2209 1330 2413 1572 1164 1491 1179 1621 1494 990 1548 2719 2324 526 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 506(S) 352 985 2207 2221 3013 1228 2099 2715 2192 3001 2167 1553 1952 1978 2279 3131 1106 1840 3241 2944 527 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 507(Y 3482 2868 3701 3919 238 3552 1112 3000 3638 2516 2526 3027 3772 3101 3341 3418 3527 3071 441 4711 528 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 508(E) 2641 3308 896 3732 3966 2458 2043 4105 2128 4016 3555 1531 2959 1842 2560 2479 2750 3722 3563 3385 529 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 509(S) 1981 864 2260 1969 2205 1291 1697 1605 1808 2070 1333 1448 1910 1601 1958 2200 636 877 2550 2211 530 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 510(F) 3342 2776 4026 4232 4354 3545 1431 2315 4038 1801 1900 3299 3780 3350 3645 3490 3420 2566 739 349 531 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 511(V 929 722 3094 2519 671 72 1536 1450 2197 1669 235 2159 2584 1873 2107 1688 882 1742 1406 1072 532 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 512(R 1152 2148 1103 556 2568 841 2681 2297 448 2220 1446 714 2063 63 2686 1086 1067 1962 2250 1802 533 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 513(V 1261 1032 2990 401 1324 3032 2128 1904 2354 591 293 2430 3023 2199 2472 2185 1232 2780 2221 1813 534 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 514(V 1755 1298 4332 3970 1769 4057 3755 2730 3843 618 544 3732 3880 3701 3920 3374 1748 2759 3277 2831 535 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 515(D 1718 3250 3403 27 3066 1736 2667 3486 1211 3381 2732 400 2240 752 1917 1439 1784 3023 3204 2299 536 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 516(S) 635 1657 1163 252 2614 1382 780 2338 601 2421 1582 466 1801 415 1101 2397 1307 1804 2665 2044 537 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 517(M 2406 2296 3638 3594 1525 3105 2824 1047 3121 596 5043 3293 3425 3046 2996 2911 2552 1398 2513 2207 538 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 518(M 2437 1975 4738 4148 487 4369 3139 1386 3872 2524 2731 4019 3789 2984 3525 3603 2318 374 2113 2174 539 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 519(T) 504 959 1413 1049 1473 1580 994 629 943 1256 601 1702 1931 837 1224 750 1993 1780 1865 1432 540 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 520(Q 2562 2904 1886 1971 3251 2661 2079 3690 1565 3469 3081 2107 3091 4371 1665 2585 2674 3411 3077 2821 541 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 521(H 2992 2589 3064 3043 863 3512 4371 2521 2584 2051 1961 2327 3545 2255 2492 2815 2919 2563 158 3265 542 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 522(F) 1777 758 2605 2069 2426 2201 1138 17 1776 51 1943 1762 2314 1474 1762 1320 786 23 1063 575 543 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 523(K 2620 2961 2461 2046 3743 2791 1570 3603 3784 3387 2839 2048 3039 1260 465 2604 2536 3331 3001 2988 544 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 524(W 3358 2599 4116 4259 2442 3892 536 2247 3821 1683 1712 2792 3835 2843 3352 3227 3271 2441 5485 1363 545 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 525(L) 841 1175 3469 2926 668 3037 2058 1328 2615 2287 379 2642 2978 2226 2526 2188 1355 301 1760 1526 546 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 526(R 837 2176 602 1392 2587 1688 216 2257 1534 2141 1281 1107 1779 227 2109 709 741 1854 2229 1683 547 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 527(M 888 1677 357 850 1922 1428 1639 1611 1435 1654 1807 103 1521 338 129 358 369 1259 1902 1285 548 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 528(A 2576 902 2349 2158 2756 1202 1937 2410 2031 2688 1852 1502 1892 1801 2162 1022 1811 1644 3004 2709 549 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 529(K 583 1746 490 34 1989 1543 1732 1666 1763 1714 880 1553 1641 238 31 516 513 875 1945 1367 550 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 1300 7108 769 894 1115 701 1378 * * 530(R 915 1387 1141 641 1786 1421 246 1638 712 1598 1052 657 1663 43 3215 962 890 1418 1542 1280 551 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 38 5845 6888 894 1115 238 2716 * * 531(E) 1710 3547 2209 3007 3749 1634 990 3679 1227 3559 2898 238 2166 662 2065 1376 1779 3163 3720 2744 552 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 532(I) 599 507 2385 1791 457 2112 969 1604 1499 1009 366 1553 2152 765 1506 1178 1103 1145 1019 667 553 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 533(P) 787 1572 724 108 1851 1418 201 1534 126 1626 789 169 1640 198 358 575 413 1199 1916 922 554 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 534(W 4114 3274 4214 4453 1722 3496 2681 4109 4149 3619 3615 4044 3885 4032 3816 4355 4248 4133 6191 1329 555 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 535(R 1940 2623 2163 1317 2718 2498 2633 2821 557 2576 1920 1294 2549 305 3406 1859 1734 2575 2279 1885 556 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 536(H 1345 2440 1207 576 2915 2072 3063 2527 2588 2343 1569 727 2129 1381 532 1222 1187 2200 2324 1939 557 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 537(D 695 2182 1890 241 2491 1456 249 2246 1496 2187 1300 85 1264 1306 385 545 648 1804 2357 1656 558 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 538(Y 1138 876 3345 2810 645 2852 1674 1928 2488 317 10 2411 2835 2166 2393 1984 1094 2117 1423 2681 559 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 539(P) 1031 974 2031 1918 2925 1207 1878 2641 1913 2865 2004 1407 2879 1695 2096 1838 694 1790 3123 2802 560 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 540(H 30 439 11 363 394 739 699 414 463 757 501 286 577 639 165 23 111 312 697 25 561 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 541(S) 1737 944 2392 2415 3092 1199 2214 2801 2395 3086 2226 1614 1940 2116 2445 2898 735 1863 3322 3065 562 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 542(M 1968 1576 4331 3761 611 3859 2765 1741 3480 2227 2250 3502 3515 2812 3229 3049 1885 1013 2038 1963 563 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 543(N 2171 2655 1458 1748 3334 2364 2267 3943 2365 3936 3437 4205 2932 2205 2608 2224 2439 3392 3253 2909 564 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 544(I) 1257 1007 3424 2868 1397 2811 1254 2188 2500 1333 382 2341 2749 1996 2283 1922 1186 115 815 2061 565 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 545(I) 1831 1391 4327 3871 1200 3991 3265 2941 3675 1302 22 3631 3727 3275 3598 3245 1791 1578 2650 2410 566 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 546(E) 999 1712 787 1203 1974 1412 152 1661 194 1721 863 101 1546 266 305 466 419 822 1983 1349 567 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 547(T) 590 1479 974 206 2293 1346 555 1995 272 2077 1218 407 1671 167 745 1655 1771 1514 2333 1741 568 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 548(S) 897 1462 2333 2543 3185 1640 2474 3294 2686 3497 2780 1973 2360 2483 2703 3465 1316 2413 3310 3025 569 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 549(T) 866 1511 1211 1054 1813 1680 2857 1969 793 2130 1455 1074 2117 914 1013 989 3041 1609 2119 1469 570 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 550(W 587 608 2405 1898 479 1928 993 42 1552 528 62 1554 2129 1332 1550 365 609 2511 3028 502 571 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 551(W 947 1346 3135 2759 3172 2759 620 882 2412 912 526 2088 2831 1938 2273 1901 1498 870 3648 929 572 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 552(Q 1772 2604 1870 1079 3217 2374 569 2777 660 2529 1835 1110 2420 3075 2723 1671 1571 2504 2437 2192 573 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 553(Q 2562 2904 1886 1971 3251 2661 2079 3690 1565 3469 3081 2107 3091 4371 1665 2585 2674 3411 3077 2821 574 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 554(D 1791 3663 3414 1659 3853 1647 1043 3807 1343 3679 3054 251 2201 725 2227 1438 1872 3282 3836 2831 575 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 555(H 3205 3079 2723 2890 2110 3046 5295 4135 2617 3813 3561 2886 3482 2833 2620 3291 3356 3895 2397 1681 576 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 556(N 2171 2655 1458 1748 3334 2364 2267 3943 2365 3936 3437 4205 2932 2205 2608 2224 2439 3392 3253 2909 577 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 557(G 2594 2690 3304 3623 4328 3747 3462 4761 3953 4671 4212 3320 3352 3748 3779 2839 2981 4004 3668 4222 578 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 558(Y 3620 2706 4179 4428 3303 4051 393 2536 4005 1939 1984 2749 3933 2855 3452 3299 3498 2688 350 3843 579 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 559(S) 344 978 2187 2155 2953 1227 2035 2643 2108 2926 2093 1523 1940 1902 2216 2907 1859 1800 3181 2875 580 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 560(H 3205 3079 2723 2890 2110 3046 5295 4135 2617 3813 3561 2886 3482 2833 2620 3291 3356 3895 2397 1681 581 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 561(Q 2562 2904 1886 1971 3251 2661 2079 3690 1565 3469 3081 2107 3091 4371 1665 2585 2674 3411 3077 2821 582 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 562(D 1724 3448 3411 103 3726 1642 1058 3734 1342 3628 2991 1884 2196 745 2191 1408 1822 3197 3743 2768 583 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 563(P) 2931 2878 3420 3706 4181 2925 3468 4621 3859 4490 4165 3491 4225 3781 3695 3182 3279 4087 3594 4064 584 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 564(G 468 1108 1987 2183 3307 3093 2261 3159 2489 3374 2507 1593 2035 2163 2570 1410 895 2119 3471 3217 585 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 565(V 371 671 3209 2618 1562 2580 1515 1368 2275 648 269 2195 2574 1917 2132 1689 852 2127 1337 1001 586 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 566(T) 1238 1032 3270 2717 705 2883 1871 1700 2401 1666 301 2442 2848 2072 2345 2015 1957 452 1675 1395 587 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 567(T) 307 965 2051 1918 2899 1272 1866 2617 1903 2838 1975 1405 1889 1683 2089 1577 2739 1773 3100 2780 588 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 568(H 837 578 1703 1139 579 1822 2864 45 902 418 1836 1086 1915 740 1071 867 423 1342 1041 648 589 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 569(L) 2092 1719 4323 3846 712 3925 2989 451 3531 2641 428 3624 3640 2952 3339 3191 2044 1542 2221 2096 590 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 570(L) 1056 648 2733 2160 1291 2277 874 5 1827 1649 341 1760 2303 1464 1729 1359 733 2 647 1605 591 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 571(E) 994 2497 67 2352 2818 1564 437 2572 1391 2487 1645 1818 1831 24 503 812 956 2133 2633 1933 592 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 572(K 1592 2196 1883 1179 2203 2360 733 1718 3129 666 1212 1218 2435 383 258 1621 1460 1669 2199 1837 593 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 971 7108 1051 894 1115 701 1378 * * 573(C 39 2488 473 1144 1043 1085 142 526 67 851 125 185 1325 126 296 170 1058 295 1316 837 594 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 360 6167 2274 894 1115 1874 459 * * 574(F) 1315 1110 2234 2129 3522 2099 247 203 1900 58 60 1632 2283 1510 1804 1624 1357 396 303 1334 595 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 38 5845 6888 894 1115 2039 402 * * 575(H 354 1360 326 436 1031 1026 2689 1410 436 1450 724 1667 1303 383 89 286 351 1096 1280 501 596 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 38 5845 6888 894 1115 238 2716 * * 576(K 1086 2304 1032 362 2797 1910 283 2399 2361 2235 1413 1695 1959 1479 1362 963 952 2031 2264 1826 597 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 577(P) 1766 2832 2091 503 3634 1897 1480 3629 1677 3608 2962 830 3445 1206 2322 1624 1926 3087 3516 2951 598 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 578(H 480 1902 1434 926 2176 1399 2271 1932 300 1903 1004 39 1521 347 782 362 423 1511 2083 1389 599 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 579(Y 839 383 2642 2032 1206 2056 863 223 1678 569 461 1617 2100 1333 1555 1135 472 1465 747 1987 600 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 580(I) 865 836 3163 2646 1108 2692 1893 2677 2382 556 157 2324 2771 2135 2380 1845 504 1519 1868 1491 601 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 581(R 1066 2251 943 347 2740 308 313 2349 1342 2216 1401 854 1954 119 2761 953 949 1986 2272 1823 602 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 582(E) 370 640 961 1875 770 1558 279 900 284 670 84 547 1598 160 606 577 301 965 907 668 603 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 583(Y 3618 2707 4172 4418 2748 4047 395 2537 3997 1940 1986 2748 3932 2853 3448 3297 3496 2688 348 4199 604 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 584(F) 2597 2122 4519 4116 2769 4202 2114 140 3797 2428 469 3639 3772 2912 3436 3471 2488 783 1265 532 605 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 585(P) 1615 1136 2222 2310 3082 1373 2227 2731 2358 3036 2272 1689 3445 2139 2424 761 938 1937 3279 3042 606 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 586(A 2397 935 2082 1846 2575 1236 1722 2190 1737 2475 1666 1377 1554 1552 1929 527 1772 1532 2826 2490 607 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 587(D 2784 3432 4016 1200 4140 2466 2197 4505 2621 4365 3956 1551 3014 2039 3232 2593 2938 4046 3710 3552 608 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 588(A 3098 930 2580 2583 2894 1243 2249 2309 2448 2781 2022 1697 1976 2194 2449 591 1113 1602 3196 2952 609 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 589(N 2171 2655 1458 1748 3334 2364 2267 3943 2365 3936 3437 4205 2932 2205 2608 2224 2439 3392 3253 2909 610 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 590(M 387 854 1643 1199 1437 1461 1020 962 1012 1269 2610 1075 1857 895 1259 1806 1865 709 1810 1406 611 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 591(H 30 439 11 363 394 739 699 414 463 757 501 286 577 639 165 23 111 312 697 25 612 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 592(L) 2625 2145 4613 4174 1968 4304 2386 72 3866 2741 551 3801 3814 2959 3500 3585 2511 745 1483 874 613 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 593(L) 2193 1789 4457 3877 523 4057 2894 279 3584 2588 2231 3689 3624 2841 3310 3261 2096 1093 2048 2035 614 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 594(A 2970 925 2555 2528 2849 1243 2202 2276 2388 2737 1975 1675 1968 2140 2406 585 1607 1581 3151 2898 615 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 595(V 1334 1034 3629 3155 1390 3214 2516 2107 2918 591 331 2852 3211 2701 2928 2407 1824 2364 2380 1985 616 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 596(M 998 406 1988 1399 1077 46 692 73 1142 733 1731 1227 1918 903 1209 286 359 167 855 487 617 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 597(H 1095 2732 1291 2125 2985 1512 3032 2799 427 2721 1890 1256 1861 159 1055 875 1088 2329 2902 2089 618 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 598(H 971 1917 1026 424 2119 1866 2995 1811 2402 1852 1093 602 1947 34 266 932 871 294 1999 1507 619 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 599(C 1836 3931 2905 2499 1252 1823 1705 1152 2189 888 389 1893 2246 1917 2150 1063 756 157 1855 1509 620 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 600(F) 528 777 2899 2354 2441 2415 823 127 2008 1363 215 1876 2433 1604 1876 1506 889 142 490 1736 621 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 601(K 1239 2390 1266 522 2934 2031 324 2495 2422 2300 1501 979 2062 1504 1964 1116 1081 2146 2296 1915 622 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 602(S) 699 1151 903 440 1699 1408 579 1323 350 1526 1574 974 1694 269 750 2107 464 1002 1909 1408 623 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 603(Q 873 2060 736 198 2478 1732 282 2118 1944 2060 1235 426 1836 1968 230 780 1853 1758 2189 1681 624 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 604(N 1080 2639 214 1668 2924 1545 527 2711 282 2635 1807 2572 1867 2104 814 876 1062 2261 2801 2043 625 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 605(K 457 1622 593 29 1532 1520 184 1170 1745 1320 528 275 1605 1259 206 467 609 916 1665 1434 626 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 606(I) 1995 1561 4353 3926 1045 4065 3261 3346 3652 924 94 3716 3783 3240 3554 3362 1957 705 2551 2295 627 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 607(N 2171 2655 1458 1748 3334 2364 2267 3943 2365 3936 3437 4205 2932 2205 2608 2224 2439 3392 3253 2909 628 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 608(Y 1025 487 2483 1892 336 2087 935 224 1576 1270 403 1587 2140 1281 1535 1165 537 999 908 2074 629 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 609(M 797 587 3073 2468 1091 2423 1329 2169 2112 34 2364 2036 2429 1745 1959 1522 645 1209 1165 836 630 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 610(V 1573 1179 4043 3576 867 3627 2822 2175 3353 312 198 3256 3486 3047 3296 2841 1540 2804 2430 2026 631 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 611(A 2440 912 2339 2201 2840 1287 1997 2505 2116 2780 1935 1522 1901 1868 2234 517 1851 1699 3081 2799 632 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 612(S) 437 1083 2006 2144 3262 1583 2197 3097 2380 3307 2434 1562 2008 2077 2485 2965 857 2074 3426 3160 633 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 613(K 2620 2961 2461 2046 3743 2791 1570 3603 3784 3387 2839 2048 3039 1260 465 2604 2536 3331 3001 2988 634 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 614(H 1501 2478 747 589 1969 2033 3845 2617 5 2483 1816 796 2272 2881 187 1387 1442 2314 2088 1345 635 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 615(P) 2931 2878 3420 3706 4181 2925 3468 4621 3859 4490 4165 3491 4225 3781 3695 3182 3279 4087 3594 4064 636 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 616(R 1242 1347 1254 731 2201 1510 719 1792 153 1944 1159 782 1844 375 2580 656 1294 1393 2232 1788 637 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 617(P) 1448 1164 798 305 1387 1533 406 980 151 81 453 488 1572 1113 529 556 448 759 1638 1144 638 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 618(Q 557 1175 1028 563 1488 1618 611 820 317 1222 557 696 1835 2724 638 698 1378 1031 1782 1308 639 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 619(W 3603 2685 4198 4439 2092 4053 373 2561 4004 1981 2004 2736 3927 2842 3443 3284 3472 2688 5224 2885 640 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 620(F) 1507 1226 3587 3073 2681 3022 1217 950 2707 1659 313 2490 2927 2137 2464 2143 1430 380 687 1671 641 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 621(S) 344 978 2188 2158 2955 1227 2038 2646 2111 2929 2096 1524 1940 1904 2218 2919 1829 1801 3183 2878 642 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 622(I) 1092 874 3179 2611 721 2765 1721 1891 2310 1192 241 2311 1836 1986 2249 1880 1042 1298 1570 1257 643 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 623(D 891 2456 2161 1856 2735 1474 410 2519 186 2456 1593 91 1753 1291 767 707 782 2062 2639 1876 644 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 624(E) 2641 3308 896 3732 3966 2458 2043 4105 2128 4016 3555 1531 2959 1842 2560 2479 2750 3722 3563 3385 645 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 625(A 3438 1472 2846 3040 3287 1726 2735 2840 3028 3257 2662 2236 2447 2798 2944 1216 1387 2183 3405 3320 646 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 626(K 719 2060 577 1623 2444 1622 164 2125 1699 2035 1164 266 1703 282 1624 601 634 1722 2150 1583 647 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 627(Q 1243 1432 728 405 2358 1377 690 2055 351 2150 1310 555 1744 2880 751 1081 618 1561 2401 1849 648 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 628(H 780 1536 555 1361 1393 1709 2249 1189 144 1529 640 494 1865 169 489 816 724 1022 1690 1099 649 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 629(C 456 2634 3043 2479 719 2386 1463 624 2148 1022 146 2055 2467 1831 2047 1515 777 2135 1370 1020 650 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 630(D 59 1947 1795 1154 2254 1392 111 1999 273 1959 1053 30 1526 335 1177 370 709 1564 2142 1456 651 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 631(Q 1161 1938 287 164 2274 1471 154 1990 1104 1956 1072 1604 1598 1906 101 461 513 1579 2132 1496 652 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 632(G 2594 2690 3304 3623 4328 3747 3462 4761 3953 4671 4212 3320 3352 3748 3779 2839 2981 4004 3668 4222 653 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 633(L) 2002 1175 2785 2475 1055 2185 1898 221 2172 2054 104 2097 2542 1964 2186 1458 1162 325 1982 1669 654 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 634(K 943 1709 403 133 2096 781 142 1801 994 1811 930 144 1543 292 938 844 405 1399 2023 1402 655 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 635(I) 525 452 2111 23 445 1963 805 1776 1266 283 345 1346 2019 1026 1323 1017 436 1518 942 1179 656 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 636(W 1626 1321 3570 3141 2053 3057 937 2580 2760 226 54 2426 2999 2185 2514 2189 1555 448 3586 581 657 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 637(D 1498 3029 3115 17 3354 1691 852 3199 1912 3109 2371 341 2120 495 1230 1252 1519 2742 3208 2450 658 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 638(W 3358 2599 4116 4259 2442 3892 536 2247 3821 1683 1712 2792 3835 2843 3352 3227 3271 2441 5485 1363 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1115 701 1378 * * 703(D 1718 3250 3403 27 3066 1736 2667 3486 1211 3381 2732 400 2240 752 1917 1439 1784 3023 3204 2299 725 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 704(E) 1165 2746 1435 2588 3046 1525 639 2845 567 2796 1988 154 1909 258 1213 943 1174 2380 2996 2179 726 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 705(E) 921 2774 1237 2767 3077 1530 662 2877 603 2830 2029 161 1924 284 1256 967 1205 2412 3032 2209 727 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 706(F) 3342 2776 4026 4232 4354 3545 1431 2315 4038 1801 1900 3299 3780 3350 3645 3490 3420 2566 739 349 728 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 707(D 1198 2653 2928 116 3164 1523 794 2975 806 2952 2173 1307 1973 435 1474 1014 1273 2473 3161 2340 729 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 708(N 497 2045 1492 265 2357 1412 1608 2108 128 2074 1183 1928 1592 229 403 517 553 1673 2265 1565 730 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 709(Y 1550 1282 3519 3004 2 3093 1385 1232 2635 1579 299 2537 2990 2153 2464 2220 1477 195 891 3290 731 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 710(F) 3342 2776 4026 4232 4354 3545 1431 2315 4038 1801 1900 3299 3780 3350 3645 3490 3420 2566 739 349 732 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 711(T) 1213 1674 2755 2906 3163 1922 2659 2698 2788 3105 2612 2311 2600 2708 2753 1463 3819 2197 3286 3156 733 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 712(K 921 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701 1378 * * 800(I) 1200 1080 2894 2252 618 2712 1476 2020 1547 1701 361 2078 2664 1554 1445 1820 1129 84 1503 1217 822 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 801(Q 1195 1999 1275 287 2307 1399 132 2058 888 2010 1104 1022 1544 1594 271 396 475 1616 2187 1494 823 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 802(E) 464 1834 313 1601 2133 1426 72 1849 1475 301 936 76 1522 1237 64 359 488 1446 2019 1377 824 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 803(H 1285 1168 1100 753 1895 1421 3671 1567 616 1783 1019 796 1815 585 942 599 1325 1196 2122 1628 825 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 804(H 457 1111 796 240 1415 1595 1499 796 56 1014 275 1464 1677 16 1113 558 395 539 1363 816 826 149 500 233 43 381 399 106 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472 91 1024 1483 125 1548 1811 1610 769 180 147 288 950 516 413 1222 1868 1282 845 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7108 8150 894 1115 701 1378 * * 824(G 736 2137 965 180 2586 1581 401 2350 145 2321 1448 1563 712 15 699 528 738 1882 2516 1799 846 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 276 7108 2582 894 1115 701 1378 * * 825(L) 1714 1375 3901 3421 618 3530 2616 734 3109 2342 497 3185 3329 2647 2982 2766 1674 1950 2019 1825 847 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 19 6850 7893 894 1115 1215 813 * * 826(K 1321 2321 787 481 2892 1879 447 2552 3186 2406 1676 717 2074 54 419 1212 1230 2209 2369 1979 848 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 1043 6850 983 894 1115 1215 813 * * 827(V 11 281 1388 1014 772 1145 782 672 779 271 145 834 1527 694 967 363 1648 1687 1388 977 849 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 38 5845 6888 894 1115 2039 402 * * 828(D 878 1818 3239 428 2222 972 438 2180 575 2276 1711 95 1458 189 1148 733 979 1822 2152 1652 850 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 38 5845 6888 894 1115 2039 402 * * 829(K 453 1667 328 1484 2007 1088 173 1659 1862 1641 861 236 1304 570 510 325 390 1317 1793 1239 851 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 38 5845 6888 894 1115 2039 402 * * 830(K 207 1058 256 150 1533 1073 83 1043 1817 1222 493 14 1295 442 505 228 1229 760 1520 1048 852 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 38 5845 6888 894 1115 2039 402 * * 831(G 414 272 462 426 1782 1704 635 1518 577 1737 936 257 1039 416 850 1592 98 853 1974 1547 853 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 38 5845 6888 894 1115 2039 402 * * 832(M 1337 339 1188 761 497 1179 525 361 511 26 2356 679 1477 431 716 350 116 415 1109 709 854 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 38 5845 6888 894 1115 2039 402 * * 833(V 753 590 2446 1973 226 2358 1433 1299 1657 1429 779 1837 2356 1469 1726 1538 740 2084 1336 966 855 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 38 5845 6888 894 1115 2039 402 * * 834(T) 413 229 707 500 1469 500 543 1022 421 1349 599 312 1063 330 674 1483 1911 534 1729 1303 856 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 38 5845 6888 894 1115 2039 402 * * 835(A 2597 310 1033 1004 1499 618 950 768 944 1290 768 669 1235 852 1081 19 107 412 1828 1482 857 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 38 5845 6888 894 1115 2039 402 * * 836(T) 93 414 954 868 1353 741 817 616 701 1116 632 643 1304 696 854 136 2822 336 1700 1317 858 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 38 5845 6888 894 1115 2039 402 * * 837(A 2597 310 1033 1004 1499 618 950 768 944 1290 768 669 1235 852 1081 19 107 412 1828 1482 859 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 38 5845 6888 894 1115 2039 402 * * 838(A 2597 310 1033 1004 1499 618 950 768 944 1290 768 669 1235 852 1081 19 107 412 1828 1482 860 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 38 5845 6888 894 1115 2039 402 * * 839(T) 93 414 954 868 1353 741 817 616 701 1116 632 643 1304 696 854 136 2822 336 1700 1317 861 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 38 5845 6888 894 1115 2039 402 * * 840(A 2597 310 1033 1004 1499 618 950 768 944 1290 768 669 1235 852 1081 19 107 412 1828 1482 862 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 38 5845 6888 894 1115 2039 402 * * 841(G 461 838 976 1119 2120 3148 1219 2101 1326 2273 1700 986 1505 1212 1430 653 781 1558 1923 1928 863 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 38 5845 6888 894 1115 2039 402 * * 842(D 878 1818 3239 428 2222 972 438 2180 575 2276 1711 95 1458 189 1148 733 979 1822 2152 1652 864 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 38 5845 6888 894 1115 2039 402 * * 843(N 431 1158 164 47 1545 910 407 1680 257 1848 1242 3201 1391 207 570 456 599 1296 1696 1106 865 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 38 5845 6888 894 1115 2039 402 * * 844(E) 771 1700 518 2837 2056 1033 298 1811 123 1925 1338 65 1445 9 532 658 819 1519 1994 1511 866 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 38 5845 6888 894 1115 238 2716 * * 845( ) 30 439 11 363 394 739 699 414 463 757 501 286 577 639 165 23 111 312 697 25 867 * * * * * * * * * * * * * * * * * * * * * * * * * * * * 0
(329) TABLE-US-00016 TABLE 7 HMMER2.0 [2.3.2] NAME XFP N ACC PF09364.1 DESC XFP N-terminal domain LENG 396 ALPH Amino RF no CS no MAP yes COM hmmbuild -f -F HMM fs.ann SEED.ann COM hmmcalibrateseed 0 HMM fs.ann NSEQ 6 DATE Thu May 3 17:57:17 2007 CKSUM 7893 GA 15.1 15.1 TC 15.1 15.1 NC 14.6 14.6 XT 8455 4 1000 1000 8455 4 8455 4 NULT 4 8455 NULE 595 1558 85 338 294 453 1158 197 249 902 1085 142 21 313 45 531 201 384 1998 644 EVD 10.948357 0.672333 A C D E F G H I K HMM m->m m->i m->d i->m i->i d->m d->d b->m m->e L M N P Q R S T V W Y 193 * 3000 1(K) 1968 2951 2155 1320 3390 2747 966 3007 2330 2844 2086 1422 2165 549 1590 1872 1791 2712 2821 1937 1 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 1193 9626 2(I) 1769 1476 4090 3550 1526 3579 2618 2387 3249 1441 544 1396 3532 2933 3155 2723 1726 1550 2406 2063 2 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9624 3(S) 1378 2318 1402 1145 3726 2032 1821 3503 1805 3583 2748 1309 2587 1497 2321 2452 2280 2794 3796 3178 3 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9622 4(D) 2415 4334 2560 2263 4523 2295 1677 4467 1966 4327 3682 889 2279 1352 2856 2064 2494 3929 4515 3478 4 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9620 5(E) 818 2294 768 2387 2689 1890 629 2403 181 2389 1504 596 2035 193 1173 879 979 1984 2580 1944 5 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9618 6(Y) 868 2568 747 2575 2297 2202 1201 2582 1098 2680 1956 1001 2523 938 1573 1484 1571 2270 2610 2844 6 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9617 7(L) 2772 2280 5308 4845 1620 5006 4218 1675 4673 2505 404 4676 4560 4047 4494 4304 2713 1658 3269 3192 7 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9615 8(H) 899 2380 623 1169 2703 1834 2073 2447 1918 2389 1478 1301 1952 64 563 794 860 2005 2553 1878 8 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9613 9(K) 1161 2683 1251 212 2990 727 723 2756 1972 2690 1794 1663 2115 1586 911 1006 1116 2301 2857 2139 9 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9611 10(M) 2709 2269 5162 4623 1349 4724 3702 1512 4374 1792 3396 4390 4320 3675 4116 3951 2630 1553 2866 2812 10 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9609 11(H) 2820 4339 3411 925 4078 2615 3698 4807 2416 4617 4088 1308 3181 1809 3221 2491 2928 4293 4210 3302 11 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9607 12(K) 1531 2609 1521 881 3185 677 855 2809 2197 2705 1875 1069 2428 423 1698 1376 1380 2418 2783 2320 12 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9605 13(Y) 4639 3574 5042 5388 3053 4919 1116 3531 4943 2851 2938 3536 4775 3673 4311 4166 4491 3684 3607 3791 13 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9604 14(W) 3826 3700 3776 3690 1600 3954 2357 4085 2609 3642 3464 3450 4249 2161 2584 3858 3853 4045 5840 1155 14 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9602 15(R) 4488 4181 4789 4318 5193 4148 3436 5568 2393 5152 4748 4156 4479 3286 4202 4614 4467 5273 4267 4649 15 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9600 16(A) 3208 1704 3999 4070 3276 2242 3400 1516 3824 2767 2340 2844 2960 3485 3660 1604 1654 1301 3916 3651 16 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9598 17(C) 3231 3331 4290 4524 4051 2061 3603 3558 4177 4038 3241 2842 2850 3725 3864 1443 1636 2656 4310 4227 17 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9596 18(N) 2093 3624 1842 565 4191 2225 1624 4071 1786 4008 3287 3436 2749 1290 2544 1859 1312 3511 4199 3291 18 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9594 19(Y) 4605 3570 5021 5357 2115 4893 1140 3492 4921 2816 2906 3546 4765 3679 4305 4160 4468 3656 389 4545 19 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9592 20(L) 3485 2916 5860 5269 1134 5582 4270 1416 5040 2720 2646 5288 4721 3890 4562 4907 3336 1219 2912 3094 20 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9591 21(A) 2093 1633 3496 3406 3943 1228 3043 3705 3256 3935 3021 2418 2652 2942 3281 2007 1343 2649 4148 3944 21 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9589 22(I) 1858 1782 4736 4347 2337 4240 3903 2415 4179 1341 1167 4002 4207 4002 4206 3513 2172 2258 3603 3167 22 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9587 23(G) 1257 1874 3207 3523 4341 3341 3408 4248 3853 4463 3557 2605 2866 3402 3760 1410 1718 3031 4475 4358 23 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9585 24(M) 2521 2386 3846 3279 1155 3984 2657 686 2594 1335 3429 3250 3784 3078 2603 3140 2428 1237 2461 2296 24 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9583 25(I) 2891 2390 5370 4957 1628 5096 4334 3554 4751 1008 437 4806 4650 4141 4574 4462 2842 63 3311 3180 25 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9581 26(Y) 4614 3571 5028 5366 3123 4901 1136 3497 4929 2819 2909 3545 4768 3679 4308 4163 4474 3661 384 4150 26 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9579 27(L) 4119 3542 5389 5357 2027 4767 4358 1609 5118 3293 979 5230 4771 4474 4724 5069 4106 2331 3412 3400 27 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9577 28(R) 1752 2663 1994 1206 3071 2612 927 2620 1567 1194 1815 1326 2642 523 2318 1048 1606 2354 2690 2328 28 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9576 29(D) 2163 3991 2853 1466 4198 2225 1494 4086 1627 3968 3237 1901 2710 1142 2410 971 2206 3573 4159 3203 29 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9574 30(N) 2100 2812 1725 2071 4541 1400 2845 4689 3101 4717 3995 3948 3237 2675 3479 2189 2459 3750 4448 4114 30 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9572 31(P) 2845 2996 4029 4118 2821 3470 3585 2547 3827 1069 2145 3734 3854 3778 3737 3186 3115 2756 3564 3278 31 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9570 32(L) 3571 3027 4990 4839 362 4751 2053 1386 4381 2878 762 3969 4502 3586 4013 4093 3476 1986 1288 2490 32 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9568 33(M) 2211 1924 4481 3865 1786 3872 2670 469 3517 2158 2560 3505 3670 2910 3263 3004 1361 903 2139 1963 33 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 1891 7944 464 894 1115 701 1378 9818 9566 34(S) 87 526 885 918 1797 699 997 1679 970 1948 1267 663 1335 874 1147 2826 252 1082 2036 1577 34 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 34 6087 7129 894 1115 2832 218 9818 9564 35(V) 734 692 2391 2193 909 2005 1781 1094 1922 136 10 1902 2325 1852 1974 1413 858 2960 1857 1405 35 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 34 6087 7129 894 1115 2832 218 9818 9562 36(T) 84 586 1220 1166 1618 901 1090 915 1004 1405 917 886 1488 990 1134 317 3114 605 1941 1587 36 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 34 6087 7129 894 1115 133 3510 9818 9560 37(R) 2583 3338 2968 1938 4226 2024 1216 3597 1648 3282 2594 1884 3156 798 2920 2479 2342 3324 3144 3007 37 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9558 38(E) 1678 2848 628 2969 3815 1219 1598 3602 1552 3614 2817 1072 2634 1251 2123 1592 1327 3012 3808 3091 38 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9557 39(P) 3058 3915 1871 1726 4973 2986 2783 5207 3189 5075 4546 2051 3849 2577 3857 2918 3276 4561 4629 4315 39 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9555 40(L) 3389 2846 5732 5157 1795 5405 3964 1554 4913 2673 66 5067 4642 3819 4457 4697 3249 1211 2753 2720 40 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9553 41(K) 1121 2502 1004 1296 2888 2042 588 2583 2027 2485 1598 681 2119 1586 1299 1000 1054 2164 2595 2010 41 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9551 42(P) 784 2052 818 1262 2296 1869 546 1970 134 107 1190 567 1773 132 1208 813 814 1637 2305 1705 42 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9549 43(E) 2121 3903 1652 3001 4126 2231 1448 3991 1462 3872 3127 825 2693 1089 2171 1828 2151 3493 4045 3137 43 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9547 44(H) 1900 3602 2493 392 3825 2165 3314 3680 1250 3583 2790 1715 2572 1675 1911 1645 1910 3197 3754 2893 44 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9545 45(I) 2547 2059 5152 4751 2029 4889 4388 2502 4626 1616 797 4548 4567 4264 4595 4202 2516 2260 3637 3383 45 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9543 46(K) 4125 4091 4017 3615 5101 3960 2958 5182 3972 4825 4345 3552 4249 2709 1809 4154 4044 4895 4132 4389 46 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9541 47(H) 826 2035 963 419 2397 1898 3051 2073 206 2136 1290 675 932 235 1162 866 1249 1718 2394 1810 47 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9539 48(R) 2700 3449 3065 1955 4296 3222 1177 3628 1592 3280 2601 1619 3187 756 3400 2568 2401 3384 3125 3003 48 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9537 49(L) 2172 1972 4362 3884 1289 3737 2942 312 3536 2565 217 3515 1361 3084 3377 2952 2169 1219 2509 2319 49 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9535 50(V) 2641 2155 5210 4776 1803 4918 4261 1765 4623 2179 584 4583 4545 4128 4515 4219 2597 2215 3420 3258 50 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9533 51(G) 4088 3924 4774 5139 5615 3825 4753 6303 5453 6014 5662 4812 4539 5232 5106 4370 4472 5527 4696 5561 51 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9531 52(H) 4836 4275 4356 4629 3663 4242 5422 5932 4492 5436 5298 4579 4696 4620 4352 5005 5030 5648 3750 3259 52 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9529 53(W) 5598 4432 5385 5722 3380 4524 4140 5941 5643 5318 5354 5427 4938 5474 5178 5909 5751 5890 6275 2993 53 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9527 54(G) 4088 3924 4774 5139 5615 3825 4753 6303 5453 6014 5662 4812 4539 5232 5106 4370 4472 5527 4696 5561 54 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9526 55(T) 1093 1706 3487 3693 4074 1966 3302 3842 3638 4134 3253 2558 2748 3296 3520 1110 3646 2760 4304 4125 55 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9524 56(C) 1052 2901 3250 2679 1060 2526 1634 1315 2341 866 266 2229 2670 2018 2238 1521 1154 1349 1596 1246 56 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9522 57(P) 1362 1972 3107 3390 4250 2166 3327 4213 3641 4411 3553 2613 3840 3308 3600 1256 1811 3082 4372 4198 57 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9520 58(G) 4088 3924 4774 5139 5615 3825 4753 6303 5453 6014 5662 4812 4539 5232 5106 4370 4472 5527 4696 5561 58 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9518 59(Q) 2629 2340 4433 3873 1140 4292 3047 1389 3342 2319 20 3746 4000 2559 3238 3469 2536 946 2531 2422 59 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9516 60(S) 1129 1869 2034 2027 3771 1956 2322 3546 2277 3697 2821 2528 2607 2062 2583 2544 1447 2665 3893 3470 60 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9514 61(F) 3253 2755 5063 4895 3979 4679 2230 1673 4579 697 843 4029 4483 3735 4216 4008 3205 1123 1463 453 61 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9512 62(I) 2667 2178 5235 4799 1779 4947 4278 2326 4646 2166 559 4613 4558 4130 4529 4251 2621 1716 3410 3264 62 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9510 63(Y) 4030 3324 4830 4953 82 4664 1400 2548 4444 776 2024 3603 4588 3589 4024 4010 3953 2880 668 4492 63 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9508 64(A) 2914 1624 3741 3790 3997 1911 3253 3736 3609 4009 3103 2541 2685 3231 3506 1407 1356 2665 4233 4079 64 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9506 65(H) 4836 4275 4356 4629 3663 4242 5422 5932 4492 5436 5298 4579 4696 4620 4352 5005 5030 5648 3750 3259 65 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9504 66(C) 868 2955 3843 3261 1152 3113 2129 1511 2905 1928 237 2799 3137 2521 2739 2241 1449 4 1921 1610 66 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9502 67(N) 3642 3838 3004 3356 4743 3612 3773 5643 4099 5460 5060 4378 4185 3865 4225 3744 3950 5002 4391 4381 67 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9500 68(R) 4488 4181 4789 4318 5193 4148 3436 5568 2393 5152 4748 4156 4479 3286 4202 4614 4467 5273 4267 4649 68 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9498 69(I) 687 1693 4550 4054 1797 4053 3239 2451 3802 1595 714 3702 3939 3479 3713 3246 2027 1554 2897 2565 69 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9496 70(I) 3202 2787 5004 4996 2836 4384 4440 3937 4838 1815 1871 4704 4600 4683 4704 4353 3280 603 3938 3661 70 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9494 71(N) 777 2760 1484 853 3260 2383 822 2877 1407 2738 1911 2481 2455 385 2206 1436 1435 2500 2791 2332 71 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9492 72(K) 1654 2912 1393 667 3173 2287 1055 2863 3038 286 2070 954 2512 658 811 1519 1598 2533 3029 2434 72 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9490 73(Y) 3965 3303 4553 4498 2089 4562 2685 3175 3880 2696 2630 3283 4469 3249 1301 3756 3844 3248 415 3893 73 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9488 74(D) 2647 4396 3720 698 4634 2448 1896 4723 2212 4559 3990 1090 3022 1848 3057 2298 2751 4181 4595 3647 74 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9486 75(Q) 786 1836 2085 1707 1785 2491 1572 1296 1287 1558 860 1741 2731 3104 1481 1664 1471 1312 2370 1975 75 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9484 76(N) 2189 3961 2351 476 4203 2257 1501 4079 1323 3955 3231 3154 2736 1150 2219 1890 2229 3577 4120 3208 76 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9482 77(M) 1639 1552 3944 3485 1584 3112 2598 102 3126 813 4345 2996 3316 2834 3037 2326 1195 1340 2475 2167 77 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9480 78(I) 2541 2130 4989 4447 1768 4503 3473 2245 4181 1829 210 4162 4181 3554 3947 3708 2469 1402 2759 2626 78 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9478 79(Y) 4613 3570 5027 5365 3096 4900 1135 3496 4928 2818 2908 3544 4767 3678 4307 4162 4473 3660 383 4168 79 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9476 80(M) 2401 1938 4995 4588 2135 4683 4177 2411 4441 955 3356 4340 4447 4169 4437 3971 2375 2371 3610 3272 80 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9474 81(C) 924 2595 3097 2552 1083 823 1551 560 2221 963 2359 2072 2516 1903 2136 739 963 2007 1573 1229 81 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9472 82(G) 4088 3924 4774 5139 5615 3825 4753 6303 5453 6014 5662 4812 4539 5232 5106 4370 4472 5527 4696 5561 82 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9470 83(P) 4497 4117 4899 5251 5560 4139 4798 6328 5454 5976 5741 5026 4302 5328 5104 4798 4844 5739 4665 5484 83 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9468 84(G) 4088 3924 4774 5139 5615 3825 4753 6303 5453 6014 5662 4812 4539 5232 5106 4370 4472 5527 4696 5561 84 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9466 85(H) 4836 4275 4356 4629 3663 4242 5422 5932 4492 5436 5298 4579 4696 4620 4352 5005 5030 5648 3750 3259 85 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9464 86(G) 4088 3924 4774 5139 5615 3825 4753 6303 5453 6014 5662 4812 4539 5232 5106 4370 4472 5527 4696 5561 86 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9461 87(G) 2125 1842 3631 3927 4350 3048 3538 4169 4033 4428 3528 2733 2876 3575 3846 1489 1707 2984 4482 4443 87 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9459 88(P) 2640 3260 2127 2297 4320 2987 2661 4510 2216 4379 3792 2411 3576 2864 2373 2683 2863 3935 4069 3806 88 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9457 89(A) 2982 1594 3754 3796 3529 1052 3162 2818 3604 3423 2664 2573 2719 3219 3468 1317 1463 1086 3887 3678 89 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9455 90(M) 1348 1165 3573 2973 1525 706 1766 1561 2606 377 3758 2527 2921 2210 2439 2009 1294 310 1556 1184 90 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9453 91(V) 1974 1692 4373 3885 1655 3760 3023 236 3586 1658 599 3493 3751 3250 3485 2959 1156 2628 2723 2419 91 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9451 92(S) 1951 1569 3422 3255 3372 1033 2819 2999 3074 3315 2500 2366 2636 2776 3104 2094 1382 1224 3674 3414 92 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9449 93(N) 1058 2388 785 572 3066 1969 1036 2804 709 2811 1947 2272 2144 1706 1182 1128 1239 2319 3013 2367 93 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9447 94(C) 968 2761 3158 2743 1848 2096 2020 1352 2456 835 1054 2166 2572 2189 2458 2381 1213 1124 2305 1969 94 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9445 95(Y) 4608 3570 5023 5360 2380 4895 1138 3494 4924 2817 2907 3545 4766 3679 4306 4161 4470 3658 386 4475 95 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9443 96(L) 3428 2867 5794 5267 1216 5534 4333 1535 5028 2951 6 5264 4750 3972 4604 4907 3306 978 2993 3105 96 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9441 97(D) 2629 4604 3026 2782 4770 2374 1828 4782 2235 4614 4052 973 2960 1523 3217 2241 2730 4225 4757 3689 97 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9439 98(G) 2862 3768 1871 1615 5004 3370 2705 5181 3146 5060 4493 1931 3470 2488 3861 2740 3106 4452 4685 4318 98 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9437 99(T) 1078 1695 3486 3659 4077 1954 3276 3853 3599 4130 3239 2539 2733 3255 3500 2681 2838 2756 4301 4124 99 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9435 100(Y 3992 3300 4836 4941 66 4664 1423 2456 4436 992 1925 3615 4579 3584 4019 4006 3913 2810 690 4437 100 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9433 101(S) 1032 1663 3422 3512 4087 1390 3188 3870 3492 4107 3184 2468 2688 3124 3454 2496 2388 2740 4293 4121 101 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9430 102(E 1823 3131 875 2933 3614 2362 1174 3322 553 3205 2407 1048 1540 771 1382 1665 1779 2906 3277 2712 102 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9428 103(F) 863 914 3349 2737 2502 2631 1388 298 2350 698 39 2234 2667 1962 2166 1719 1033 1141 1185 1869 103 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9426 104(Y 4352 3468 4770 5008 2112 4760 1124 3396 4660 2798 2831 1505 4661 3557 4156 4005 4236 3515 387 4271 104 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9424 105(P) 1085 2001 3515 3791 4319 2231 3514 4118 3884 4378 3566 2802 3892 3541 3758 1662 1872 3068 4416 4363 105 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9422 106(N 1628 3254 1441 2076 3527 2085 1091 3331 913 3246 2403 2397 2414 688 1516 1414 1613 2859 3417 2622 106 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9420 107(I) 1569 2626 3973 3416 1377 3324 2311 2751 3082 941 528 2965 3319 2753 2944 2457 1527 1473 2109 2078 107 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9418 108(T 908 1834 1592 1168 2951 1885 1379 2651 1086 2760 1898 1245 2295 1915 1496 1739 2152 2098 3019 2494 108 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9416 109(M 879 2216 787 1054 1375 1857 485 2215 1265 2203 1871 512 1943 1655 1154 781 811 1824 2400 1773 109 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9414 110(D 2264 3415 3677 1019 4473 2407 2124 4366 2457 4415 3784 1339 3017 1850 3214 2128 1551 3724 4492 3732 110 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9412 111(E 1628 2756 1092 2433 3004 2358 986 2615 1561 1281 1850 1034 2517 587 533 1527 1543 2348 2851 2339 111 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9409 112(E 800 2932 1355 2151 3219 1985 890 3005 643 2935 2058 1450 2244 1611 1220 1179 1330 2541 3109 2350 112 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9407 113(G 4088 3924 4774 5139 5615 3825 4753 6303 5453 6014 5662 4812 4539 5232 5106 4370 4472 5527 4696 5561 113 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 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117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9386 123(F) 4150 3318 4992 5163 3777 4801 1232 2695 4739 954 2105 3556 4637 3592 4183 4031 4023 3011 484 2534 123 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9384 124(P) 4497 4117 4899 5251 5560 4139 4798 6328 5454 5976 5741 5026 4302 5328 5104 4798 4844 5739 4665 5484 124 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9382 125(G 4088 3924 4774 5139 5615 3825 4753 6303 5453 6014 5662 4812 4539 5232 5106 4370 4472 5527 4696 5561 125 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9379 126(G 4088 3924 4774 5139 5615 3825 4753 6303 5453 6014 5662 4812 4539 5232 5106 4370 4472 5527 4696 5561 126 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9377 127(I) 2878 2414 5081 4778 2015 4697 2956 3488 4533 580 695 4245 4462 3882 4297 4026 2840 329 2190 1350 127 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9375 128(P) 1112 1752 3015 3242 4210 2077 3186 4033 3529 4251 3331 2415 3138 3114 3525 1101 1561 2858 4391 4208 128 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9373 129(S) 1944 2458 3696 4011 4410 2644 3753 4675 4191 4807 4070 3198 3384 3886 4051 3656 2393 3615 4374 4307 129 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9371 130(H 4836 4275 4356 4629 3663 4242 5422 5932 4492 5436 5298 4579 4696 4620 4352 5005 5030 5648 3750 3259 130 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9369 131(M 726 2516 3320 2699 1540 2560 1464 135 2311 587 3158 2207 2612 1942 2131 1653 952 1415 1331 989 131 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9366 132(A 2437 1736 2547 2496 3668 1942 2557 3365 2590 3601 2745 1872 2625 2364 2796 1269 2286 2528 3861 3523 132 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9364 133(P) 2199 1920 3558 3810 4266 2163 3478 4045 3859 4314 3481 2754 3455 3500 3727 1576 1785 2985 4394 4324 133 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9362 134(E 2302 4193 1609 3106 4369 2263 1582 4283 1777 4150 3462 851 2781 1870 2594 1969 2359 3762 4338 3341 134 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9360 135(T 1509 1748 4032 3964 2803 2637 3374 581 3680 2093 1801 3053 3237 3460 3596 1989 3160 2274 3655 3308 135 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9358 136(P) 4497 4117 4899 5251 5560 4139 4798 6328 5454 5976 5741 5026 4302 5328 5104 4798 4844 5739 4665 5484 136 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9355 137(G 4088 3924 4774 5139 5615 3825 4753 6303 5453 6014 5662 4812 4539 5232 5106 4370 4472 5527 4696 5561 137 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9353 138(S) 1402 1678 3661 3893 4185 1940 3408 3979 3851 4254 3335 2598 2733 3421 3678 3267 1525 2799 4407 4277 138 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9351 139(I) 2917 2413 5393 4971 1597 5119 4331 3518 4766 1140 403 4828 4654 4126 4574 4483 2863 105 3287 3176 139 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9349 140(H 2691 3511 1351 1619 2597 2858 4977 4322 2002 4099 3606 1623 3363 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5389 5357 2027 4767 4358 1609 5118 3293 979 5230 4771 4474 4724 5069 4106 2331 3412 3400 149 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9326 150(S) 1113 1682 3645 3884 4190 1942 3410 3986 3853 4261 3343 2598 2736 3423 3680 3340 1530 2804 4411 4280 150 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9324 151(H 2323 2561 3097 3086 1647 3098 4888 2143 2537 2630 2333 2801 3547 2728 2559 2581 2516 1445 2255 1333 151 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9322 152(A 2928 1667 3720 3939 4168 1934 3412 3952 3863 4230 3311 2605 2728 3429 3681 2146 1516 2782 4394 4269 152 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7944 8986 894 1115 701 1378 9818 9320 153(Y 3656 3073 4655 4705 2072 4415 1160 2834 4311 2480 2370 3332 4386 3385 3897 3630 1330 2897 449 4200 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500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8807 344(K 2113 3150 2249 1388 3797 2829 992 3281 2710 3026 2265 1725 2831 1860 1625 1985 1911 2964 2962 2675 346 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8804 345(P) 2272 3432 4625 4892 4644 2928 4139 4609 4669 4819 4180 3725 3909 4404 4368 2543 2718 3748 4454 4686 347 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8801 346(E 2460 4036 460 3371 4348 2457 1702 4275 1584 4118 3462 1104 2946 2051 2090 2167 2503 3804 4163 3385 348 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8798 347(E 2181 3963 1851 3002 4213 2236 1529 4092 1674 3990 3269 843 2732 1182 2455 1882 1292 3581 4185 3235 349 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8794 348(L) 3686 3084 5725 5318 2066 5451 3323 791 5050 2922 90 4961 4743 3882 4541 4834 3541 1590 2260 1671 350 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8791 349(F) 3772 3147 5332 5198 4004 5042 2246 1256 4859 1259 621 4264 4667 3786 4369 4411 3663 1931 1412 419 351 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8788 350(D 2611 4416 3402 640 4705 2395 1885 4789 2297 4633 4071 2681 2988 1589 3255 2256 2733 4207 4711 3684 352 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8785 351(E 2436 2868 924 2744 4080 2309 2015 3815 2053 3929 3209 1413 2875 1718 2576 1853 2106 3206 4090 3471 353 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8781 352(H 692 2743 2175 223 3038 1936 2515 2810 480 2751 1863 1409 2154 1594 1031 1057 1179 2357 2927 2196 354 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8778 353(G 2929 3168 3518 3782 1975 3489 2803 4394 4019 4184 3856 3400 3940 3733 3967 3094 3258 3950 2519 2220 355 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8775 354(S) 743 1505 1390 843 1747 1981 884 1345 674 588 830 999 2148 615 1093 1645 1025 1123 2016 1540 356 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8771 355(P) 2511 2204 4688 4170 1213 4215 3207 1516 3824 2221 98 3895 2348 3271 3624 3424 2456 677 2586 2462 357 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8768 356(K 1302 1414 2419 1840 1577 2693 1530 1325 2435 1189 660 1855 2761 1446 1616 1749 1313 1240 2101 1697 358 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8765 357(E 687 2542 623 1957 2905 1944 710 2641 1433 2592 1707 604 1914 274 748 998 1082 2205 2761 2094 359 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8762 358(E 2040 3771 1647 2163 4056 1964 1435 3921 1508 3821 3060 809 2658 1075 2248 864 2081 3410 4011 3098 360 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8758 359(I) 2526 2113 4983 4447 1598 4503 3494 2404 4186 1749 242 4162 4188 3576 3962 3711 2456 1420 2790 2649 361 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8755 360(R 1519 2765 1433 1043 3230 2362 786 2853 1407 2712 1878 1010 2424 346 2271 1399 2090 2477 2766 2295 362 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8752 361(A 2266 3817 1712 2253 4133 2221 1507 3996 1626 3910 3174 845 2706 1158 2386 1834 2166 3485 4107 3184 363 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8748 362(F) 1042 885 3196 2582 2439 2571 2436 1238 2213 706 10 2148 2609 1856 2072 1653 1024 245 1243 832 364 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8745 363(M 2115 1084 3069 2501 1009 2553 1611 411 2186 950 2374 2167 2680 1891 2150 1666 1057 397 1620 1292 365 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8741 364(P) 4497 4117 4899 5251 5560 4139 4798 6328 5454 5976 5741 5026 4302 5328 5104 4798 4844 5739 4665 5484 366 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8738 365(K 1362 2752 891 1110 3142 2149 769 2837 2560 2727 1865 782 1373 1618 441 1221 1281 2420 2829 2241 367 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8735 366(G 1022 1909 2732 2555 3858 2953 2403 3572 1903 3702 2863 2142 2736 2152 1711 1431 1592 2728 3841 3572 368 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8731 367(E 1926 3649 1536 2753 3890 2165 1327 3735 1325 3640 2851 1567 2586 953 2024 1665 1092 3244 3826 2947 369 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8728 368(K 2153 2373 3052 2232 1767 3262 1649 1215 3030 859 2514 2219 3217 1370 831 2425 2024 1487 2493 2241 370 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8725 369(R 4488 4181 4789 4318 5193 4148 3436 5568 2393 5152 4748 4156 4479 3286 4202 4614 4467 5273 4267 4649 371 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8721 370(M 3269 2747 5652 5079 1177 5325 4102 1525 4829 1255 4460 5012 4616 3829 4428 4616 3145 974 2895 3004 372 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8718 371(G 2030 1651 3665 3855 4195 2704 3377 3993 3822 4242 3302 2568 2705 3371 3670 1117 1491 2786 4410 4294 373 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8714 372(A 2364 1861 1953 1763 3466 1952 1976 3199 1760 3331 2471 1666 2522 1987 2080 1936 1374 2468 3555 3102 374 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8711 373(N 1618 2562 1583 1250 3475 2356 1296 3134 474 3082 2296 3378 2657 908 1726 1605 1129 2676 3159 2725 375 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8707 374(P) 1913 2274 2279 2086 1517 2783 2782 2081 1589 560 1612 2058 3317 1785 1716 2097 1976 2000 2063 1260 376 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8704 375(H 929 2010 791 1272 2044 1913 2963 1853 298 1959 1145 1363 2029 239 766 888 872 799 2186 1512 377 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8700 376(A 2877 1669 3800 3976 4022 1962 3382 3596 3795 3999 3156 2631 2746 3409 3616 1328 2512 2627 4284 4151 378 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8697 377(N 3122 3352 2328 2577 596 3446 1619 3701 2981 3309 3133 3888 3831 2693 3130 3026 3247 3561 1270 2585 379 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8693 378(G 2018 2692 1922 2271 4544 3363 2969 4678 3249 4731 3990 1984 3232 2825 3564 2140 2400 3690 4460 4184 380 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8690 379(G 1257 1874 3207 3523 4341 3341 3408 4248 3853 4463 3557 2605 2866 3402 3760 1410 1718 3031 4475 4358 381 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8687 380(Y 1236 1100 3220 2594 757 2737 1473 388 2061 1865 99 2233 2760 1870 1132 1823 1172 1244 1376 2076 382 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8683 381(L) 2379 2141 1254 3407 1292 3975 2923 1518 3405 2543 203 3359 3842 2943 3392 3194 2319 538 2579 2378 383 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8679 382(R 1224 1996 1644 1058 2408 2207 1005 2022 378 315 1393 1189 1416 662 2668 937 1215 1747 2459 2006 384 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 288 7945 2519 894 1115 701 1378 9818 8676 383(R 1347 2594 872 1389 3097 2078 754 2768 37 2671 1846 784 2259 333 2876 1137 1287 2364 2765 2230 385 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7671 8713 894 1115 1555 600 9818 8672 384(N 1422 3052 1734 1461 3326 980 901 3136 759 3056 2213 1864 2210 500 1385 1028 1413 2660 3236 2429 386 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7671 8714 894 1115 345 2232 9818 8669 385(L) 4119 3542 5389 5357 2027 4767 4358 1609 5118 3293 979 5230 4771 4474 4724 5069 4106 2331 3412 3400 387 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8665 386(K 1622 2790 1624 1030 3260 2461 828 2846 2708 2716 1905 1116 2506 393 1496 1511 1487 892 2768 2339 388 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8662 387(M 1888 1623 4198 3600 1092 3572 2482 158 3255 2140 2397 3210 3470 2776 3059 2699 1034 1479 2114 1890 389 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8658 388(P) 4497 4117 4899 5251 5560 4139 4798 6328 5454 5976 5741 5026 4302 5328 5104 4798 4844 5739 4665 5484 390 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8655 389(D 2522 4504 3422 1446 4683 938 1750 4661 2127 4504 3910 909 2894 1438 3092 2147 2619 4102 4695 3603 391 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8651 390( 3715 3061 5054 5000 2784 4779 1574 1778 4602 1668 1132 3738 4524 3553 4110 4005 3593 2304 4571 289 392 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8647 391(H 1137 2529 1169 405 2911 2042 2148 2611 1514 2512 1626 678 2128 1722 2034 1013 1053 2189 2623 2031 393 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8644 392(Q 1523 3117 2166 1365 3403 2057 1004 3194 1438 3110 2253 667 2351 2526 1325 1325 1499 2728 3276 2509 394 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 9818 8640 393(Y 4598 3559 5006 5329 2257 4898 2594 3515 4887 2848 2926 3523 4759 3654 4280 4143 4453 3662 366 4237 395 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8988 894 1115 701 1378 9818 8637 394(A 2380 2381 1061 1282 3846 2045 1941 3614 1958 3704 2890 1385 2665 1636 2457 1509 1729 2895 3915 3319 396 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8988 894 1115 701 1378 9818 8633 395(V 2097 1741 4406 4026 2222 4103 3555 2399 3840 1319 1108 1444 4071 3681 3882 3349 2094 2762 3366 2920 397 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8988 894 1115 701 1378 9818 8629 396(D 928 2371 1886 165 2715 1822 563 2464 1387 2421 1512 478 1080 116 692 750 1043 2018 2599 1913 398 * * * * * * * * * * * * * * * * * * * * * * * * * * * 9818 0
(330) TABLE-US-00017 TABLE 8 HMMER2.0 [2.3.2] NAME XFP ACC PF03894.6 DESC D-xylulose 5-phosphate/D-fructose 6-phosphate phosphoketolase LENG 187 ALPH Amino RF no CS no MAP yes COM hmmbuild -f -F HMM fs.ann SEED.ann COM hmmcalibrate --seed 0 HMM fs.ann NSEQ 6 DATE Sun Apr. 29 15:58:33 2007 CKSUM 8559 GA 15.5 15.5 TC 18.2 15.7 NC 13.6 14.2 XT 8455 4 1000 1000 8455 4 8455 4 NULT 4 8455 NULE 595 1558 85 338 294 453 1158 197 249 902 1085 142 21 313 45 531 201 384 1998 644 EVD 10 339054 0.664450 A C D E F G H I K HMM m -> m m -> i m -> d i -> m i -> i d -> m d -> d b -> m m -> e L M N P Q R S T V W X Y 222 * 2807 1(M) 919 1308 2565 2222 1930 1975 1841 1394 1946 1641 3218 1876 1316 1826 2064 1219 2730 1183 2403 2047 1 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7717 8760 894 1115 701 1378 1222 8539 2(R) 691 1720 837 294 1975 1756 501 673 144 1751 937 1196 1878 121 1881 951 681 1325 2078 1516 2 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7717 8760 894 1115 701 1378 8762 8535 3(V) 885 1621 999 990 1982 1862 915 1514 733 1796 1048 873 1044 622 1147 858 916 2279 2227 1717 3 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7717 8760 894 1115 701 1378 8762 8531 4(L) 2842 2379 5209 4634 994 4802 3633 174 4364 2574 2538 4478 4252 3491 4019 4045 2735 1455 2614 2661 4 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7717 8760 894 1115 701 1378 8762 8527 5(G) 1046 1667 2864 3162 4077 3372 3114 3979 3511 4193 3299 2337 2643 3089 3461 1104 1501 2797 4216 4066 5 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7717 8760 894 1115 701 1378 8762 8524 6(K) 696 1959 661 115 2242 1707 387 1932 1754 1971 1104 400 1809 1727 429 726 657 544 2214 1595 6 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7717 8760 894 1115 701 1378 8762 8520 7(Y) 4363 3343 4802 5127 2939 4674 927 3251 4692 2586 2669 3329 4545 3458 4081 3935 4227 3413 177 4223 7 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7717 8760 894 1115 701 1378 8762 8516 8(C) 1627 2687 3970 3367 1521 3283 2148 37 3007 2149 118 2930 3201 2520 2790 2405 1562 1206 1785 1498 8 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7718 8760 894 1115 701 1378 8762 8512 9(R) 737 2550 1594 497 3149 1980 968 2870 526 2846 2019 774 2281 570 3052 1221 1340 2417 3004 2383 9 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7718 8760 894 1115 701 1378 8762 8508 10(D) 2399 4358 3324 2308 4527 2166 1608 4527 1994 4367 3790 765 2746 1301 2656 2018 2496 3976 4515 3456 10 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7718 8760 894 1115 701 1378 8762 8504 11(V) 2284 1804 4895 4504 1990 4629 4189 2412 4382 1011 761 4283 4358 4111 4394 3938 2259 2686 3549 3219 11 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7718 8760 894 1115 701 1378 8762 8500 12(M) 2278 1867 4746 4224 1303 4284 3343 1656 3973 1240 3304 3939 3999 3423 3784 3494 2217 2073 2691 2538 12 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7718 8760 894 1115 701 1378 8762 8496 13(K) 693 2202 1423 749 2553 2182 674 2107 2480 2170 1388 928 2252 275 1432 1210 1146 1012 2378 1932 13 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7718 8760 894 1115 701 1378 8762 8492 14(N) 1049 1434 1773 1500 1636 2109 1397 1127 1366 1815 824 2287 2427 1298 1590 1307 1405 1004 2112 1674 14 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7718 8760 894 1115 701 1378 8762 8488 15(N) 3232 3514 2565 2905 4358 3267 3364 5192 3627 5059 4632 4348 3843 3410 3792 3319 3533 4568 4087 3976 15 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 867 7718 1171 894 1115 701 1378 8762 8484 16(E) 1220 2441 99 2757 2966 1568 808 2774 675 2772 2036 363 2075 466 1190 1070 1278 2337 2879 2244 16 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 22 6873 7915 894 1115 2169 363 8762 8480 17(D) 1004 1749 2891 305 2104 1632 964 886 981 1762 1266 575 2040 692 1495 1057 1089 1706 2499 1874 17 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 460 6873 1933 894 1115 216 2846 8762 8476 18(M) 823 1578 1383 173 1837 1620 411 1480 107 1624 2041 425 938 38 569 595 1047 1187 1968 1402 18 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7493 8536 894 1115 1355 715 8762 8472 19(T) 897 2121 888 291 2541 1825 413 2206 1465 2164 1310 532 1923 20 1428 1124 1894 1823 2313 1770 19 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7493 8536 894 1115 404 2032 8762 8468 20(N) 867 2024 881 423 2654 1782 693 2361 230 2371 1499 2664 2000 273 1178 870 1294 1909 2569 1974 20 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7718 8760 894 1115 701 1378 8762 8464 21(F) 3465 2876 5057 4893 3938 4762 2035 1021 4547 1305 409 4001 4411 3532 4090 4106 3362 1663 1215 228 21 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7718 8760 894 1115 701 1378 8762 8459 22(R) 4070 3868 4406 3890 4820 3837 3026 5122 1946 4739 4314 3731 4155 2844 4177 4178 4041 4836 3949 4248 22 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7718 8760 894 1115 701 1378 8762 8455 23(I) 2379 1901 4955 4531 1728 4662 4059 2900 4381 1570 512 4321 4336 3965 4313 3959 2342 1650 3306 3087 23 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7718 8760 894 1115 701 1378 8762 8451 24(F) 2676 2354 4538 4394 3992 3956 1878 728 4074 869 959 3497 4006 3383 3780 3277 2708 1235 1189 154 24 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7718 8760 894 1115 701 1378 8762 8447 25(G) 1031 1653 2874 3165 4069 3287 3106 3964 3500 4180 3282 2329 2632 3078 3451 1430 1486 2782 4212 4060 25 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7718 8760 894 1115 701 1378 8762 8443 26(P) 4077 3791 4518 4858 5206 3818 4460 5898 5053 5602 5343 4628 4289 4938 4749 4364 4429 5313 4385 5119 26 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7718 8760 894 1115 701 1378 8762 8439 27(D) 3832 4189 4132 2236 5073 3341 3194 5633 3716 5365 5035 2584 3886 3102 4252 3662 4000 5151 4434 4544 27 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7718 8760 894 1115 701 1378 8762 8435 28(E) 3777 4125 1925 3871 4984 3355 3125 5379 3371 5156 4796 2611 3875 3007 3762 3630 3916 4960 4357 4463 28 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7718 8760 894 1115 701 1378 8762 8430 29(T) 1009 1466 1761 1492 1840 2028 1429 1343 1350 766 1045 1393 2396 1299 1587 1239 2910 1154 2256 1801 29 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7718 8760 894 1115 701 1378 8762 8426 30(M) 1499 1643 917 1243 1962 1786 691 1586 425 1763 2853 693 1975 349 862 779 773 1304 2139 1601 30 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7718 8760 894 1115 701 1378 8762 8422 31(S) 1644 2173 3321 3615 4077 2357 3411 4307 3792 4459 3725 2859 3095 3512 3692 3619 2088 3285 4091 3959 31 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7718 8760 894 1115 701 1378 8762 8418 32(N) 3232 3514 2565 2905 4358 3267 3364 5192 3627 5059 4632 4348 3843 3410 3792 3319 3533 4568 4087 3976 32 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7718 8760 894 1115 701 1378 8762 8414 33(R) 2793 3369 3662 2108 4312 3237 1071 3580 2493 3189 2545 1934 3162 654 3328 2667 2432 3373 2988 2947 33 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7718 8760 894 1115 701 1378 8762 8409 34(L) 3410 2836 5464 5039 1728 5176 3154 570 4762 2953 115 4708 4504 3649 4284 4539 3272 1347 2103 1550 34 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7719 8761 894 1115 701 1378 8762 8406 35(W) 901 2337 1407 65 2541 1747 2100 2368 133 2334 1447 1385 1904 1561 644 770 849 1946 3634 1773 35 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8760 894 1115 701 1378 8762 8401 36(A) 2155 3109 1523 1273 3479 1112 1076 3288 1004 3231 2417 592 2329 693 1651 1350 1580 2804 3429 2615 36 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 145 7718 8760 894 1115 701 1378 8762 8397 37(V) 1023 958 3146 2600 1103 2513 1686 63 2290 801 2200 2204 2668 2000 2237 1650 1119 2408 1689 1335 37 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8760 894 1115 701 1378 8762 8392 38(F) 3852 3057 4741 4884 3335 4537 1011 2489 4461 957 1921 3313 4388 3349 3926 3759 3730 2762 269 3290 38 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8760 894 1115 701 1378 8762 8388 39(E) 2083 3395 722 2943 3878 2383 1214 3575 2285 3395 2666 1041 2696 830 750 1870 2023 3182 3392 2873 39 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8760 894 1115 701 1378 8762 8384 40(V) 1086 1419 2113 1929 1724 796 2671 1274 1766 1761 1123 1760 2533 1690 1904 1385 1234 2825 2172 1669 40 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8760 894 1115 701 1378 8762 8379 41(T) 869 1487 3137 3257 3760 1746 2945 3506 3200 3791 2920 2268 2512 2898 3156 2152 3197 2486 3990 3777 41 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8760 894 1115 701 1378 8762 8375 42(K) 1040 2442 581 1137 2735 1868 548 2490 2386 2427 1554 1311 2012 114 440 908 975 2076 2562 1837 42 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8760 894 1115 701 1378 8762 8371 43(R) 2795 3370 3663 2111 4313 3239 1074 3582 2384 3191 2548 1936 3164 657 3395 2670 2434 3375 2990 2949 43 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 291 7718 2514 894 1115 701 1378 8762 8366 44(Q) 1085 1755 1145 887 1733 985 888 1755 814 1851 1208 1026 2253 3209 1142 1159 1127 1521 1673 859 44 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7443 8484 894 1115 1459 652 8762 8362 45(W) 4735 3754 4703 4993 2377 3915 3273 4870 4782 4319 4338 4620 4317 4639 4390 4998 4881 4870 6237 1984 45 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7443 8486 894 1115 372 2136 8762 8358 46(M) 914 874 871 1922 783 2384 1220 1249 1670 662 3001 1756 2427 1420 1731 1438 1257 98 1343 990 46 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8760 894 1115 701 1378 8762 8353 47(M) 641 2071 521 1084 2362 1649 321 2091 82 2076 2071 1285 1749 1468 420 684 616 1678 2276 1612 47 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8760 894 1115 701 1378 8762 8349 48(Q) 757 2178 529 1705 2487 519 368 2219 1195 2182 1278 340 1806 1749 411 642 696 453 2362 1699 48 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8760 894 1115 701 1378 8762 8344 49(Y) 1288 1144 3267 2757 710 2784 1561 2546 2393 763 315 2345 2866 2105 2314 1913 1335 38 1365 2756 49 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8760 894 1115 701 1378 8762 8340 50(K) 1352 2460 1542 473 2590 2065 876 2234 1932 1737 1566 761 2285 505 815 1266 1304 1990 2658 2071 50 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8760 894 1115 701 1378 8762 8335 51(P) 1299 2774 236 1949 3178 1870 886 2961 678 2912 2058 1447 2218 481 1251 890 1302 2487 3099 2347 51 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 291 7718 2514 894 1115 701 1378 8762 8331 52(P) 1084 2190 341 2185 2933 1688 929 2655 732 2706 1884 590 2271 556 1245 978 1128 2169 2938 2278 52 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7443 8486 894 1115 1459 652 8762 8326 53(N) 1418 2809 2479 106 2949 1799 926 2766 883 578 2068 2542 2188 576 1502 1234 1435 2399 3048 2264 53 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 283 7443 2563 894 1115 1459 652 8762 8322 54(D) 888 2124 2400 66 2357 1565 509 1956 285 2113 1336 255 1834 1955 808 781 871 1155 2428 1753 54 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 19 7180 8222 894 1115 1865 463 8762 8317 55(D) 1805 3680 2903 2554 3868 1689 1066 3813 1341 3684 3041 289 2233 743 2210 1458 1880 3289 3847 2846 55 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 1334 7180 753 894 1115 1865 463 8762 8313 56(G) 515 887 1038 1187 2183 3187 1282 2175 1398 2342 1770 1048 1555 1281 1496 708 838 1625 1974 1993 56 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 41 5887 6930 894 1115 2652 250 8762 8308 57(G) 515 887 1038 1187 2183 3187 1282 2175 1398 2342 1770 1048 1555 1281 1496 708 838 1625 1974 1993 57 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 41 5887 6930 894 1115 151 3329 8762 8304 58(E) 1346 1562 1816 2436 2337 2525 1279 603 1533 1038 631 1602 2663 1361 1800 1658 1319 1113 1614 909 58 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8760 894 1115 701 1378 8762 8299 59(H) 933 961 2242 1707 764 2320 2094 1132 1393 1839 65 1195 2382 1248 1495 1381 874 319 1292 847 59 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8760 894 1115 701 1378 8762 8295 60(M) 1293 1342 2996 2500 1038 2617 1796 510 2162 890 3410 2240 2790 1938 2186 2108 1323 630 1840 1502 60 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8760 894 1115 701 1378 8762 8290 61(P) 2028 1663 1726 1624 3398 998 1915 3145 1828 3283 2409 1487 2266 1625 2185 1038 1207 2349 3511 3060 61 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8760 894 1115 701 1378 8762 8285 62(D) 752 2203 1771 1070 2520 1688 359 2261 1383 2210 1298 324 1796 94 1124 630 691 543 2379 1707 62 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 231 7718 2838 894 1115 701 1378 8762 8281 63(R) 796 2105 516 1238 2496 1498 421 2206 49 2188 1314 382 1834 11 1567 698 1155 1794 2375 1748 63 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7504 8546 894 1115 412 2011 8762 8276 64(G) 2419 2885 3067 2886 4406 3143 2559 4353 1679 4234 3572 2708 3407 2316 2444 2556 2650 3716 3837 3878 64 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8760 894 1115 701 1378 8762 8271 65(R) 1920 2516 2433 1527 2802 2746 961 1879 176 2261 1647 1532 2759 1746 2935 1945 1744 2067 2537 2247 65 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8760 894 1115 701 1378 8762 8267 66(V) 2252 1773 4891 4567 2335 4633 4538 1812 4482 1132 1073 4339 4440 4385 4597 3992 2250 3374 3973 3489 66 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8760 894 1115 701 1378 8762 8262 67(M) 849 923 1670 1529 893 2248 1067 233 1327 600 2147 1485 2308 1135 1498 1282 1243 1140 1394 1015 67 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8760 894 1115 701 1378 8762 8257 68(E) 1776 3015 373 3097 3872 2071 1546 3755 1570 3731 2992 934 2595 1219 2164 1764 1926 3163 3855 3091 68 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8760 894 1115 701 1378 8762 8253 69(M) 1335 1165 3272 2688 1002 2941 1834 1451 2360 376 3121 2427 2930 2286 2331 2034 1279 1529 1766 1445 69 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8760 894 1115 701 1378 8762 8248 70(L) 3777 3243 5089 5032 1735 4514 4040 1299 4754 3261 696 4891 4508 4152 4395 4705 3767 1993 3125 3069 70 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8760 894 1115 701 1378 8762 8243 71(S) 1644 2173 3321 3615 4077 2357 3411 4307 3792 4459 3725 2859 3095 3512 3692 3619 2088 3285 4091 3959 71 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8760 894 1115 701 1378 8762 8238 72(E) 3777 4125 1925 3871 4984 3355 3125 5379 3371 5156 4796 2611 3875 3007 3762 3630 3916 4960 4357 4463 72 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8760 894 1115 701 1378 8762 8234 73(H) 4406 3961 3921 4171 3231 3926 5401 5468 3992 5013 4849 4130 4378 4151 3893 4548 4592 5195 3383 2818 73 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8760 894 1115 701 1378 8762 8229 74(Q) 1172 1561 1803 1343 1477 2292 1190 991 908 1463 565 1415 2454 2378 1119 1403 1904 979 1998 1592 74 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8760 894 1115 701 1378 8762 8224 75(C) 943 4854 3669 3391 1886 2164 2449 922 3000 676 1025 2492 2705 2686 2861 1458 1303 805 2559 2242 75 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8761 894 1115 701 1378 8762 8219 76(E) 2096 3637 282 2979 3947 2215 1377 3824 1173 3688 2987 859 2659 2792 1646 1832 2117 3370 3761 3011 76 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8761 894 1115 701 1378 8762 8214 77(G) 3681 3593 4389 4747 5281 3812 4419 5908 5071 5671 5288 4417 4223 4850 4766 3952 4069 5125 4424 5214 77 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8761 894 1115 701 1378 8762 8209 78(W) 4254 3318 4804 5081 3098 4588 1050 3111 4645 2453 2534 3403 4509 3506 4073 3950 4146 3314 5352 877 78 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8761 894 1115 701 1378 8762 8205 79(L) 3375 2814 5645 5079 928 5372 4042 409 4777 2894 2856 5098 4540 3683 4317 4734 3229 1208 2698 2837 79 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8761 894 1115 701 1378 8762 8200 80(E) 2150 3708 263 3290 4011 2224 1421 3906 1260 3766 3081 868 2687 2032 1753 1875 2180 3447 3837 3070 80 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8761 894 1115 701 1378 8762 8195 81(G) 2022 1618 3291 3557 4069 3053 3239 3867 3682 4133 3245 2462 2643 3254 3542 1260 1474 2727 4214 4140 81 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7718 8761 894 1115 701 1378 8762 8190 82(Y) 4444 3646 4593 4892 509 4268 1873 3993 4663 3407 3460 3884 4503 3998 4255 4336 4491 4072 1177 4829 82 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7719 8761 894 1115 701 1378 8762 8185 83(L) 1931 1973 3870 3595 1374 3144 2842 646 3171 2519 338 3150 3415 2899 3072 2469 2215 950 2583 2364 83 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7719 8761 894 1115 701 1378 8762 8180 84(L) 3777 3243 5089 5032 1735 4514 4040 1299 4754 3261 696 4891 4508 4152 4395 4705 3767 1993 3125 3069 84 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7719 8761 894 1115 701 1378 8762 8175 85(T) 2081 2497 3792 4048 4206 2712 3671 3972 3974 4274 3765 3291 3408 3829 3824 2338 3976 3321 4129 4203 85 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7719 8761 894 1115 701 1378 8762 8170 86(G) 3681 3593 4389 4747 5281 3812 4419 5908 5071 5671 5288 4417 4223 4850 4766 3952 4069 5125 4424 5214 86 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7719 8761 894 1115 701 1378 8762 8165 87(R) 4070 3868 4406 3890 4820 3837 3026 5122 1946 4739 4314 3731 4155 2844 4177 4178 4041 4836 3949 4248 87 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7719 8761 894 1115 701 1378 8762 8160 88(H) 1602 2202 2033 2051 2524 2310 4500 3130 1754 3221 2593 1952 2860 1941 1889 1744 2351 2641 2857 2142 88 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7719 8761 894 1115 701 1378 8762 8155 89(G) 1424 1648 3257 3538 4097 3299 3253 3902 3697 4166 3283 2476 2666 3269 3560 1290 1506 2760 4228 4161 89 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7719 8761 894 1115 701 1378 8762 8150 90(F) 653 1307 3869 3274 2383 3222 2101 2307 2923 979 54 2851 3158 2477 2735 2341 1496 62 1785 1478 90 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7719 8761 894 1115 701 1378 8762 8145 91(F) 4268 3318 4807 5092 4134 4606 1026 3118 4656 2457 2538 3389 4515 3495 4075 3946 4154 3320 3520 904 91 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7719 8761 894 1115 701 1378 8762 8140 92(A) 2436 1450 3080 3062 3722 1707 2801 3478 3005 3713 2813 2174 1760 2697 3044 2014 1242 2447 3930 3709 92 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7719 8761 894 1115 701 1378 8762 8134 93(S) 797 2971 3664 3672 3546 1712 2998 3220 3388 3539 2685 2356 2477 3023 3254 2975 1394 2305 3830 3649 93 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7719 8761 894 1115 701 1378 8762 8139 94(Y) 4444 3646 4593 4892 509 4268 1873 3993 4663 3407 3460 3884 4503 3998 4255 4336 4491 4072 1177 4829 94 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7719 8761 894 1115 701 1378 8762 8124 95(E) 3777 4125 1925 3871 4984 3355 3125 5379 3371 5156 4796 2611 3875 3007 3762 3930 3916 4960 4357 4463 95 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7719 8761 894 1115 701 1378 8762 8119 96(A) 2588 1457 3368 3530 3881 1722 3095 3647 3472 3926 3021 2338 2505 3081 3355 2536 1295 2531 4108 3950 96 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7719 8761 894 1115 701 1378 8762 8114 97(F) 4350 3618 4843 5137 4483 4245 2206 3493 4990 2887 3007 4147 4512 4243 4506 4433 4439 3737 1500 417 97 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7719 8761 894 1115 701 1378 8762 8109 98(M) 706 1201 3821 3231 1007 3165 2103 1510 2884 1469 2600 2801 3121 2481 2711 2285 1405 1421 1846 1550 98 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7719 8761 894 1115 701 1378 8762 8103 99(H) 1732 2580 1961 1265 3039 2507 3869 2740 117 2656 1919 1338 2617 548 2281 1696 1373 2439 2662 2280 99 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7719 8761 894 1115 701 1378 8762 8098 100(I) 2262 1772 4916 4590 2360 4692 4608 2948 4520 1146 1086 4377 4472 4432 4647 4052 2257 2744 4024 3536 100 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7719 8761 894 1115 701 1378 8762 8093 101(V) 2259 1770 4911 4585 2361 4682 4599 2472 4513 1151 1088 4370 4467 4427 4640 4043 2254 3105 4021 3531 101 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7719 8761 894 1115 701 1378 8762 8087 102(D) 1281 2033 3087 908 2698 2028 1475 1596 1454 2381 1758 1118 2459 1191 1938 1365 1158 1286 3025 2452 102 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7719 8761 894 1115 701 1378 8762 8082 103(S) 1644 2173 3321 3615 4077 2357 3411 4307 3792 4459 3725 2859 3095 3512 3692 3619 2088 3285 4091 3959 103 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7719 8761 894 1115 701 1378 8762 8077 104(M) 3457 3250 4578 4682 2595 3839 3833 2264 4294 1708 5248 4331 4225 4184 4072 3938 3655 2653 3435 3236 104 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7719 8761 894 1115 701 1378 8762 8071 105(L) 2433 2031 4839 4270 1637 4331 3217 1476 3985 2288 96 3990 3978 3290 3710 3525 2348 1287 2465 2367 105 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7719 8761 894 1115 701 1378 8762 8066 106(N) 952 1482 1959 1737 2360 1930 1748 1480 1701 2148 1486 2935 2420 1580 1960 1182 2024 1339 2731 2304 106 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7719 8761 894 1115 701 1378 8762 8061 107(Q) 3758 3792 3072 3245 4342 3586 3274 5053 2880 4719 4410 3349 4049 4526 2916 3829 3917 4726 3973 3953 107 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7719 8761 894 1115 701 1378 8762 8055 108(H) 3941 3303 3939 4079 336 4266 4844 3317 3580 2730 2734 3097 4295 3109 3362 3663 3868 3403 373 2757 108 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7719 8761 894 1115 701 1378 8762 8050 109(A) 2381 1319 2661 2353 1521 924 2061 2159 2216 2443 1684 1875 2365 1993 2375 1314 1085 1685 2841 2472 109 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7719 8761 894 1115 701 1378 8762 8044 110(K) 3709 3782 3592 3179 4734 3639 2561 4748 3938 4426 3926 3130 3916 2292 1410 3723 3624 4465 3818 3998 110 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7719 8761 894 1115 701 1378 8762 8039 111(W) 4239 3322 4800 5070 2397 4565 1086 3101 4634 2445 2527 3425 4503 3523 4074 3959 4140 3310 5693 837 111 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7719 8761 894 1115 701 1378 8762 8033 112(L) 3174 2633 5520 4994 1029 5242 4053 1453 4737 2932 174 4959 4514 3738 4340 4596 3057 761 2778 2863 112 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7719 8761 894 1115 701 1378 8762 8028 113(K) 1073 2547 1983 1205 2885 1851 580 2624 2040 2541 1656 477 2021 141 1183 924 1014 2183 2682 2014 113 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7719 8761 894 1115 701 1378 8762 8022 114(M) 1488 784 2685 2089 758 2378 1239 1284 1794 540 2454 1834 2427 1221 1791 1444 817 1168 1289 933 114 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7719 8761 894 1115 701 1378 8762 8017 115(C) 1116 3924 3654 3615 3360 1383 2916 3025 3341 3335 2502 2336 2465 2960 3214 1051 2263 2200 3668 3472 115 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 1842 7719 489 894 1115 701 1378 8762 8011 116(V) 588 579 2160 1946 759 1834 1555 1143 1670 9 123 1684 2156 1608 1741 1222 711 2811 1682 1223 116 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 42 5919 6961 894 1115 152 3320 8762 8006 117(R) 740 1533 1479 915 1874 1953 873 1460 464 601 933 1028 2151 588 2555 747 902 1223 2081 1631 117 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7719 8761 894 1115 701 1378 8762 8000 118(E) 1519 3167 1458 2619 3424 1929 2323 3240 838 3155 2323 534 2279 1614 1462 1295 1511 2766 3331 2516 118 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7719 8761 894 1115 701 1378 8762 7994 119(I) 1108 1022 2831 2246 840 2607 1460 2037 1908 1407 36 2034 2633 1279 1920 1682 1280 126 1499 1169 119 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7719 8761 894 1115 701 1378 8762 7989 120(P) 1226 2486 584 427 3101 1037 903 2834 1189 2796 1949 1466 2619 497 899 1126 1246 2362 2960 2322 120 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7719 8761 894 1115 701 1378 8762 7983 121(W) 5218 4132 5085 5404 2925 4252 3753 5470 5268 4877 4907 5071 4661 5108 4833 5506 5369 5443 6261 2535 121 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7719 8761 894 1115 701 1378 8762 7977 122(R) 2786 3324 3159 2158 3444 3203 2835 3646 19 3307 2690 2048 3245 942 3648 2697 2534 3415 2888 2551 122 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7719 8761 894 1115 701 1378 8762 7972 123(H) 661 2311 894 302 2686 811 2065 2371 1951 2300 1425 565 1991 22 1309 873 899 1971 2431 1856 123 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7719 8761 894 1115 701 1378 8762 7966 124(D) 1783 3362 2774 330 3711 2050 1197 3534 1468 3445 2669 689 2225 827 1652 1548 1805 3056 3598 2798 124 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7719 8761 894 1115 701 1378 8762 7960 125(I) 2784 2321 4408 4214 2256 4011 1193 3098 3823 1290 1179 3131 3919 3037 3465 3182 2704 1420 513 2319 125 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7719 8761 894 1115 701 1378 8762 7954 126(S) 1918 1460 2882 2748 3664 1367 2596 3426 2688 3630 2719 2048 1372 2426 2827 1999 1206 2419 3840 3573 126 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7719 8762 894 1115 701 1378 8762 7948 127(S) 1644 2173 3321 3615 4077 2357 3411 4307 3792 4459 3725 2859 3095 3512 3692 3619 2088 3285 4091 3959 127 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7719 8762 894 1115 701 1378 8762 7943 128(I) 2843 2399 5187 4720 1139 4765 3832 83 4437 2876 38 4542 4330 3660 4145 4094 2781 1324 2793 2769 128 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7719 8762 894 1115 701 1378 8762 7937 129(N) 3232 3514 2565 2905 4358 3267 3364 5192 3627 5059 4632 4348 3843 3410 3792 3319 3533 4568 4087 3976 129 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7719 8762 894 1115 701 1378 8762 7931 130(Y) 1616 1344 3951 3366 912 3313 2164 1486 3018 1578 20 2936 3238 2574 2829 2438 1554 1298 1825 2867 130 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7719 8762 894 1115 701 1378 8762 7925 131(I) 2438 1963 4986 4538 1573 4670 3957 2309 4367 2112 365 4331 4311 3863 4246 3955 2391 1749 3152 2992 131 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7720 8762 894 1115 701 1378 8762 7919 132(M) 694 667 2636 2033 642 2235 1106 145 1725 645 2178 1733 2301 1435 1691 561 1254 1172 1134 779 132 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7720 8762 894 1115 701 1378 8762 7913 133(T) 866 1487 3132 3246 3768 1744 2940 3519 3190 3799 2925 2263 2509 2888 3152 2652 2778 2491 3995 3782 133 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7720 8762 894 1115 701 1378 8762 7907 134(S) 1644 2173 3321 3615 4077 2357 3411 4307 3792 4459 3725 2859 3095 3512 3692 3619 2088 3285 4091 3959 134 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7720 8762 894 1115 701 1378 8762 7901 135(H) 1254 2216 843 755 2687 1970 3314 2636 839 2710 1932 1437 2343 837 1206 1244 2636 2214 2810 2158 135 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7720 8762 894 1115 701 1378 8762 7895 136(W) 2217 1998 4211 3930 1050 3517 2291 446 3456 1314 1150 3322 3697 3202 3309 2906 2265 3288 3612 961 136 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7720 8762 894 1115 701 1378 8762 7889 137(W) 1105 2160 3956 3730 1959 3434 1130 1795 3348 1692 1411 2810 3571 2764 3122 2642 2381 1804 5259 566 137 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7720 8762 894 1115 701 1378 8762 7883 138(R) 2269 3123 2497 1540 3728 2880 2414 3259 347 2971 2265 1547 2868 2539 3022 2145 2019 2993 2838 2604 138 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7720 8762 894 1115 701 1378 8762 7877 139(Q) 3758 3792 3072 3245 4342 3586 3274 5053 2880 4719 4410 3349 4049 4526 2916 3829 3917 4726 3973 3953 139 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7720 8762 894 1115 701 1378 8762 7870 140(D) 2399 4357 3309 2333 4526 2166 1607 4526 1993 4365 3789 765 2745 1300 2954 2017 2495 3974 4514 3455 140 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7720 8762 894 1115 701 1378 8762 7864 141(H) 4406 3961 3921 4171 3231 3926 5401 5468 3992 5013 4849 4130 4378 4151 3893 4548 4592 5195 3383 2818 141 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7720 8762 894 1115 701 1378 8762 7858 142(N) 3232 3514 2565 2905 4358 3267 3364 5192 3627 5059 4632 4348 3843 3410 3792 3319 3533 4568 4087 3976 142 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7720 8762 894 1115 701 1378 8762 7852 143(G) 3681 3593 4389 4747 5281 3812 4419 5908 5071 5671 5288 4417 4223 4850 4766 3952 4069 5125 4424 5214 143 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7720 8762 894 1115 701 1378 8762 7845 144(F) 4352 3337 4799 5121 3888 4670 928 3235 4686 2571 2654 3328 4541 3456 4078 3931 4217 3400 178 3260 144 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7720 8762 894 1115 701 1378 8762 7839 145(T) 866 1487 3132 3246 3768 1744 2940 3519 3190 3799 2625 2263 2509 2888 3152 2652 2778 2491 3995 3782 145 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7720 8762 894 1115 701 1378 8762 7833 146(H) 4406 3961 3921 4171 3231 3926 5401 5468 3992 5013 4849 4130 4378 4151 3893 4548 4592 5195 3383 2818 146 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7720 8762 894 1115 701 1378 8762 7827 147(Q) 3758 3792 3072 3245 4342 3586 3274 5053 2880 4719 4410 3349 4049 4526 2916 3829 3917 4726 3973 3953 147 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7720 8762 894 1115 701 1378 8762 7820 148(D) 2001 3292 3601 626 4238 2111 1759 4246 2092 4199 3539 963 2715 1470 2900 1255 2206 3561 4288 3412 148 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7720 8762 894 1115 701 1378 8762 7814 149(P) 4077 3791 4518 4858 5206 3818 4460 5898 5053 5602 5343 4628 4289 4938 4749 4364 4429 5313 4385 5119 149 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7720 8762 894 1115 701 1378 8762 7807 150(G) 2415 2868 3074 2972 4434 3359 2704 4428 1915 4330 3670 2780 3430 2492 1875 2564 2676 3754 3904 3964 150 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7720 8762 894 1115 701 1378 8762 7801 151(F) 2339 1986 4665 4064 3176 4095 2859 229 3739 1224 2452 3737 3789 3040 3446 3256 2245 1216 2175 2011 151 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7720 8762 894 1115 701 1378 8762 7794 152(I) 2056 1731 4386 3851 1203 3873 2916 2392 3540 2065 105 3529 3714 3075 3381 3054 1098 51 2437 2237 152 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7720 8762 894 1115 701 1378 8762 7788 153(D) 1118 2211 2195 621 3243 803 1223 3009 1065 3036 2177 857 2246 848 1597 985 2152 2413 3246 2588 153 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7720 8762 894 1115 701 1378 8762 7781 154(H) 1003 1199 2356 1954 1388 2206 3132 636 1619 1264 667 1770 2496 1555 1734 1385 1395 2407 1879 1430 154 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7720 8762 894 1115 701 1378 8762 7775 155(M) 695 1534 4131 3535 944 3528 2430 2154 3199 1674 2687 3161 3396 2699 2998 2660 1749 182 2022 1814 155 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7720 8762 894 1115 701 1378 8762 7768 156(L) 682 1592 4101 3606 1242 3407 2684 95 3291 2495 186 3196 3443 2879 3157 2606 1799 1329 2351 2114 156 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7720 8762 894 1115 701 1378 8762 7762 157(N) 1782 3518 1546 1312 3758 1994 1175 3610 1194 3512 2730 2977 2426 804 1905 1148 1804 3115 3698 2808 157 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7720 8762 894 1115 701 1378 8762 7755 158(K) 2459 2938 2880 2055 2893 3094 1378 2442 3407 840 1882 2005 3159 1028 200 2490 2293 2474 2837 2553 158 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7720 8762 894 1115 701 1378 8762 7748 159(K) 1148 2117 1407 950 3066 1999 1021 2720 2980 2713 1884 1076 2995 623 548 886 1179 2234 2851 2379 159 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7720 8762 894 1115 701 1378 8762 7741 160(P) 1034 1295 2486 2150 1616 1809 1863 1877 2017 2165 1435 1781 2897 1813 2195 966 1055 1493 2567 2152 160 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7720 8762 894 1115 701 1378 8762 7735 161(D) 2104 3998 2881 1352 4180 1216 1393 4095 1610 3965 3275 1701 2586 1055 2452 1771 2164 3570 4160 3154 161 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 1843 7720 490 894 1115 701 1378 8762 7728 162(N) 498 1221 110 15 1620 964 477 1773 335 1934 1330 3292 1450 280 647 522 670 1383 1763 1178 162 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 43 5920 6962 894 1115 2643 252 8762 7721 163(D) 961 1901 3312 369 2315 1032 511 2292 663 2377 1817 34 1523 264 1242 811 1065 1927 2232 1739 163 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 43 5920 6962 894 1115 2643 252 8762 7714 164(H) 997 1393 635 583 386 1411 4407 1664 259 1594 1144 676 1756 466 415 1034 1055 1464 790 42 164 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 43 5920 6962 894 1115 152 3320 8762 7707 165(V) 2121 1684 4679 4254 1449 4310 3633 2372 4076 839 756 3954 4126 3800 4049 3566 2094 2791 3155 2717 165 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7720 8763 894 1115 701 1378 8762 7700 166(V) 1172 1063 3285 2755 1280 2703 1906 1464 2465 907 407 2383 2845 2193 2434 949 1115 2471 1913 1547 166 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7721 8763 894 1115 701 1378 8762 7693 167(R) 2428 3127 2310 1828 3921 2880 1372 3657 155 3392 2732 1602 3079 997 3685 2367 2329 3316 3168 2942 167 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7721 8763 894 1115 701 1378 8762 7687 168(I) 673 1102 3159 2611 1247 2870 1866 2195 2325 820 354 2364 2914 1287 2345 1977 1221 2181 1906 1534 168 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7721 8763 894 1115 701 1378 8762 7679 169(Y) 4444 3646 4593 4892 509 4268 1873 3993 4663 3407 3460 3884 4503 3998 4255 4336 4491 4072 1177 4829 169 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7721 8763 894 1115 701 1378 8762 7672 170(F) 3400 2827 5361 4990 2846 5067 2816 651 4697 2592 25 4506 4471 3608 4224 4409 3268 1403 1851 1138 170 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7721 8763 894 1115 701 1378 8762 7665 171(P) 1191 1755 3163 3402 3998 1985 3182 3756 3492 4031 3233 2502 3818 3181 3412 1407 1613 2766 4124 4023 171 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7721 8763 894 1115 701 1378 8762 7658 172(P) 1902 1437 3147 3131 3577 1724 2798 3245 3019 3540 2684 2204 3083 2726 3028 1065 1432 2336 3823 3602 172 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7721 8763 894 1115 701 1378 8762 7651 173(D) 3832 4189 4132 2236 5073 3341 3194 5633 3716 5365 5035 2584 3886 3102 4252 3662 4000 5151 4434 4544 173 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 17 7721 8763 894 1115 701 1378 8762 7644 174(A) 2913 1375 3689 3354 3146 1014 2798 2432 3176 3025 2284 2297 2499 2833 3104 1111 1235 1120 3514 3272 174 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 18 7722 8764 894 1115 701 1378 8762 7638 175(N) 3232 3514 2565 2905 4358 3267 3364 5192 3627 5059 4632 4348 3843 3410 3792 3319 3533 4568 4087 3976 175 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 18 7721 8764 894 1115 701 1378 8762 7629 176(C) 775 3036 2850 2408 1588 1919 1731 1110 2133 1469 2376 1909 2363 1881 2164 925 2536 890 2035 1692 176 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 18 7721 8763 894 1115 701 1378 8762 7622 177(L) 3406 2832 5440 5026 2063 5151 3069 587 4746 2880 97 4659 4495 3638 4269 4506 3269 1359 2041 1444 177 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 18 7721 8763 894 1115 701 1378 8762 7615 178(L) 3777 3243 5089 5032 1735 4514 4040 1299 4754 3261 696 4891 4508 4152 4395 4705 3767 1993 3125 3069 178 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 18 7721 8763 894 1115 701 1378 8762 7607 179(C) 2942 2982 3827 3914 3641 1706 3124 3330 3613 3649 2779 2414 2485 3185 3403 1168 1242 2356 3925 3779 179 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 18 7721 8763 894 1115 701 1378 8762 7600 180(V) 2004 1608 4544 4142 2135 4125 3700 1840 3967 1140 967 3830 4052 3798 4000 3390 1338 2999 3399 2963 180 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 18 7721 8763 894 1115 701 1378 8762 7592 181(F) 1588 850 2996 2447 2261 2299 1475 182 2129 756 166 2019 2485 1821 2060 1435 1085 1429 1479 1117 181 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 18 7721 8763 894 1115 701 1378 8762 7585 182(D) 1088 3814 3226 1427 4086 2058 1384 3970 1570 3872 3169 670 2570 1045 2378 1733 2102 3453 4076 3107 182 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 18 7721 8763 894 1115 701 1378 8762 7577 183(H) 2540 3259 3155 1838 4078 3083 3373 3420 2334 3069 2392 1745 3017 555 2597 2412 2222 3187 2907 2791 183 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 18 7721 8763 894 1115 701 1378 8762 7570 184(C) 998 4910 3920 3803 2550 2066 2898 917 3459 2125 1635 2602 2717 3092 3274 1392 1333 1279 3175 2885 184 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 18 7721 8763 894 1115 701 1378 8762 7562 185(M) 3195 2670 5480 4897 2597 5129 3599 426 4630 2310 2844 4767 4399 3550 4175 4399 3050 1206 2449 2343 185 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 18 7721 8763 894 1115 701 1378 8762 7555 186(Q) 1978 3010 1873 1184 3573 2640 2315 3132 2388 2890 2142 1292 2679 3064 193 1845 1787 2819 2820 2501 186 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 18 7721 8763 894 1115 701 1378 8762 7547 187(S) 1150 2122 1356 858 3376 1832 1484 3135 1404 3212 2374 1042 2342 1143 1916 2458 2054 2488 3434 2812 187 * * * * * * * * * * * * * * * * * * * * * * * * * * * * 8762 0
(331) TABLE-US-00018 TABLE 9 HMMER2.0 [2.3.2] NAME XFP C ACC PF09363.1 DESC XFP C-terminal domain LENG 212 ALPH Amino RF no CS no MAP yes COM hmmbuild -f -F HMM fs.ann SEED.ann COM hmmcalibrate-seed 0 HMM fs.ann NSEQ 6 DATE Thu May 3 17:57:02 2007 CKSUM 5491 GA 25.0 25.0 TC 47.0 29.4 NC 15.7 15.7 XT 8455 4 1000 1000 8455 4 8455 4 NULT 4 8455 NULE 595 1558 85 338 294 453 1158 197 249 902 1085 142 21 313 45 531 201 384 1998 644 EVD 10.483098 0.640397 A C D E F G H I K HMM m->m m->i m->d i->m i->i d->m d->d b->m m->e L M N P Q R S T V W Y 193 * 2999 1(K) 4125 4091 4017 3615 5101 3960 2958 5182 3972 4825 4345 3552 4249 2709 1809 4154 4044 4895 4132 4389 1 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 1193 8721 2(Q) 1392 2802 618 424 3105 2084 2223 2893 446 2828 1977 1616 1057 3114 866 1245 1350 2468 2957 2283 2 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 8914 8718 3(P) 1097 2000 3515 3791 4319 2230 3513 4117 3884 4378 3565 2801 3889 3540 3757 1661 1871 3066 4416 4363 3 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 8914 8714 4(H) 829 2314 845 285 2656 1910 2103 2366 0 2324 1434 569 2003 1714 2028 709 889 1956 2490 1870 4 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 8914 8711 5(W) 872 1301 1524 969 1324 2103 863 924 771 407 486 1096 1119 2314 1087 1098 833 778 2741 1233 5 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 8914 8707 6(Q) 1139 1910 1850 1613 3267 2003 1790 2923 1462 3080 2266 1578 2518 3569 1755 1258 1437 2329 3351 2895 6 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 8914 8704 7(W) 4606 3595 5007 5318 253 4844 1211 3574 4838 2908 2986 3590 4756 3710 4268 4192 4482 3721 5473 3368 7 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 8914 8700 8(L) 2343 2040 4542 3937 1831 4019 2749 1548 3592 2218 65 3613 3780 1482 3353 3156 2252 936 2180 1956 8 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 8914 8697 9(T) 1385 2322 1417 1149 3726 2036 1831 3500 1823 3585 2754 1314 2593 1510 2340 1921 2800 2796 3801 3184 9 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 8914 8693 10(M) 1554 1399 3470 2849 1043 3109 1939 255 1071 1028 3713 2584 3080 2156 2304 2204 1488 1337 1856 1572 10 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 8914 8690 11(D) 2255 4080 2935 2308 4312 2253 1576 4208 1768 4097 3398 854 2767 1236 2584 1939 1141 3689 4293 3313 11 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 8914 8687 12(E) 1169 2634 621 2512 2971 1972 696 2711 1193 2634 1746 602 2134 1564 675 777 1111 2271 2782 2114 12 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 8914 8683 13(A) 3176 1703 4006 4070 3246 2260 3401 1441 3823 2722 2304 2855 2973 3487 3663 1620 1659 1420 3901 3631 13 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 8914 8679 14(I) 1158 1549 1727 1889 1669 2385 1107 1890 836 1344 736 1323 2456 885 1317 1399 1097 1018 2055 1611 14 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 8914 8676 15(H) 848 2322 1179 560 2630 2119 2363 2275 2038 2278 1446 801 2188 233 1325 1089 1073 627 2469 1939 15 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 8914 8672 16(H) 1354 2628 576 1333 2749 2068 3770 2560 570 422 1777 1432 2292 513 1032 1235 1312 2213 2778 2099 16 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 8914 8669 17(C) 947 4245 3771 3246 1380 2866 2176 26 2899 843 500 2705 3040 2556 2758 2038 1382 1326 2062 1723 17 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 8914 8665 18(T) 1296 2623 1140 1900 3034 2174 690 2702 1486 2593 1730 817 2254 245 1344 1174 1932 2302 2686 2143 18 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 8914 8662 19(A) 2122 2640 1402 254 2963 1940 736 2715 1187 2660 1774 577 2132 1562 869 1024 1125 2273 2833 2136 19 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 8914 8658 20(G) 4088 3924 4774 5139 5615 3825 4753 6303 5453 6014 5662 4812 4539 5232 5106 4370 4472 5527 4696 5561 20 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 8914 8655 21(I) 1832 1779 4707 4321 2320 4169 3857 2857 4145 1329 1156 3958 4165 3960 4165 3443 2157 1658 3571 3143 21 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 8914 8651 22(A) 1979 2132 1358 1230 3624 895 1764 3399 1659 3464 2589 1343 2501 1420 2138 1914 1456 2656 3665 3086 22 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 8914 8647 23(I) 1613 1432 3370 991 1684 3301 2327 2219 2660 1128 723 2715 3318 2464 2756 2418 1220 2185 2433 2038 23 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 8914 8644 24(W) 3333 2943 4821 4705 865 4258 2319 1550 4105 1483 1550 3928 4360 3731 3856 3853 3350 1751 5765 662 24 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8987 894 1115 701 1378 8914 8640 25(D) 2155 3941 2895 1331 4165 2244 1467 4031 2266 3906 3170 838 2712 1110 2170 1857 2186 3532 4076 3168 25 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8988 894 1115 701 1378 8914 8637 26(W) 4535 3580 5043 5339 2264 4800 1315 3404 4909 2727 2820 3667 4744 3774 4331 4221 4434 3613 5781 611 26 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8988 894 1115 701 1378 8914 8633 27(A) 3609 2508 4184 4493 4592 2737 3989 4421 4496 4701 4051 3440 3474 4171 4253 2294 2487 3536 4484 4643 27 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8988 894 1115 701 1378 8914 8629 28(S) 1944 2458 3696 4011 4410 2644 3753 4675 4191 4807 4070 3198 3384 3886 4051 3656 2393 3615 4374 4307 28 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8988 894 1115 701 1378 8914 8626 29(N) 1462 2164 1894 2122 3935 2171 2578 3801 2603 4009 3230 3172 2856 2392 2846 1616 2907 2960 4077 3633 29 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8988 894 1115 701 1378 8914 8622 30(D) 2036 3092 3356 1144 4437 2344 2201 4300 2537 4380 3702 1420 2974 1935 3248 2004 2335 3577 4473 3774 30 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8988 894 1115 701 1378 8914 8618 31(K) 719 2804 1286 282 3133 1980 846 2900 2557 2836 1958 1613 2216 421 1059 1135 1263 2444 3007 2283 31 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8988 894 1115 701 1378 8914 8615 32(G) 1795 2851 1703 1000 4330 2898 2022 4220 2294 4229 3471 1259 2791 1732 2991 1182 2066 3405 4399 3633 32 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 1206 7945 837 894 1115 701 1378 8914 8611 33(N) 713 1555 300 476 2841 2208 1131 2756 1105 2843 2074 2836 1863 861 1525 760 971 2081 2888 2349 33 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 24 6764 7806 894 1115 162 3231 8914 8607 34(E) 652 2550 1383 2004 2857 1868 643 2618 298 2563 1660 497 2040 1542 832 908 985 2168 2738 2028 34 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8988 894 1115 701 1378 8914 8604 35(E) 978 3351 336 2983 3871 2170 1425 3692 1401 3640 2859 1765 2619 1063 2043 1667 1932 3175 3830 3006 35 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8988 894 1115 701 1378 8914 8600 36(P) 1823 2153 3843 3947 3271 2734 3495 1581 3785 2766 2465 3169 3446 3607 3696 2195 2168 2328 3859 3579 36 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8988 894 1115 701 1378 8914 8596 37(D) 2642 4390 3693 696 4628 2447 1891 4714 2203 4552 3980 1088 3019 1971 3045 2293 2745 4173 4588 3642 37 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8988 894 1115 701 1378 8914 8592 38(V) 2557 2122 5071 4711 1962 4673 4220 320 4512 984 791 4445 4483 4165 4454 4020 2550 3348 3513 3210 38 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7945 8988 894 1115 701 1378 8914 8589 39(V) 2838 2616 4834 4880 3213 3955 4416 610 4757 2310 2253 4383 4345 4643 4642 3732 3007 3763 4157 3886 39 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 12 7946 8988 894 1115 701 1378 8914 8585 40(M) 3038 2583 5400 4796 1866 4936 3671 2288 4513 1389 3379 4610 4379 3590 4127 4157 2917 1104 2668 2694 40 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8581 41(A) 3609 2508 4184 4493 4592 2737 3989 4421 4496 4701 4051 3440 3474 4171 4253 2294 2487 3536 4484 4643 41 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8577 42(C) 2065 2925 3988 4043 3973 1287 3331 3726 3772 3993 3088 2601 2687 3352 3601 2060 1451 2653 4224 4091 42 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8574 43(A) 2866 2846 4109 4001 2791 2206 3077 1268 3662 2419 1888 2761 2871 3275 3465 1533 1500 1398 3377 3097 43 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8570 44(G) 4088 3924 4774 5139 5615 3825 4753 6303 5453 6014 5662 4812 4539 5232 5106 4370 4472 5527 4696 5561 44 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8566 45(D) 1008 3741 3309 578 4350 2242 1698 4254 1941 4177 3484 1820 2795 1375 2757 1930 2306 3665 4371 3418 45 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8562 46(V) 1488 1444 2805 1912 1672 3036 1995 1499 2149 1163 719 2290 3083 1996 2352 2120 998 2130 2347 1936 46 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8558 47(P) 2288 2497 3856 3931 2960 3134 3483 1588 3705 2343 2229 3416 3829 3629 3645 2690 2575 1605 3649 3323 47 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8555 48(T) 1494 1874 3613 3499 2448 2472 2890 1675 3034 1763 2674 2770 3069 2947 2983 1828 3524 1602 3212 2888 48 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8551 49(F) 1077 1060 2607 2006 1617 2517 1308 1338 853 1327 74 1856 2554 1561 1724 1568 1013 416 1457 1095 49 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8547 50(E) 4205 4417 2343 3901 5349 3698 3534 5837 3838 5561 5237 3039 4218 3453 4201 4072 4352 5409 4640 4860 50 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8543 51(T) 1578 1365 3818 3310 1660 3164 2456 1709 3026 1148 726 2898 3288 2750 2975 2334 2031 1609 2396 2024 51 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8539 52(M) 3164 2670 5546 4970 1172 5167 3972 395 4712 2611 2735 4857 4533 3759 4329 4434 3047 1340 2847 2936 52 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8535 53(A) 3164 2290 2449 2301 3900 2380 2191 3516 1483 3644 2906 2112 2939 1904 1531 1776 1904 2880 3730 3462 53 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8531 54(A) 3609 2508 4184 4493 4592 2737 3989 4421 4496 4701 4051 3440 3474 4171 4253 2294 2487 3536 4484 4643 54 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8527 55(V) 950 941 2811 2248 1031 2327 1422 440 1962 740 250 1933 2494 1690 1964 896 1079 1807 1521 1165 55 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8524 56(H) 659 2340 1265 1241 2654 1788 2240 2406 89 2353 1435 421 1907 1509 604 738 910 1959 2525 1836 56 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8520 57(M) 905 1698 4314 3724 1143 3706 2623 2145 3388 1126 3388 3345 3578 2895 3189 2841 1918 295 2218 2005 57 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8516 58(L) 3094 2629 5472 5009 1335 5054 4133 266 4744 2900 148 4850 4577 3917 4430 4403 3029 1404 3024 3024 58 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8512 59(N) 1718 2924 1536 943 3401 2491 2272 3022 67 2851 2042 2480 2560 1853 2240 1589 1584 2659 2864 2426 59 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8508 60(Q) 1303 2782 604 1957 3121 2037 786 2861 1397 2771 1895 1587 2225 2403 741 1147 1246 2420 2908 2240 60 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8504 61(M) 836 1314 1097 815 1234 2045 2007 937 682 1198 2233 976 2124 1369 1042 1018 776 996 1675 1208 61 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8500 62(F) 1264 1055 3592 2980 2448 801 1770 1523 2606 732 96 2510 2888 2223 2426 1975 1207 1242 1588 1251 62 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 267 7946 2622 894 1115 701 1378 8914 8496 63(P) 4031 3755 4476 4814 5166 3783 4422 5849 5008 5560 5299 4583 4287 4894 4709 4316 4383 5266 4354 5078 63 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7693 8735 894 1115 1509 624 8914 8492 64(E) 1699 3425 1700 2340 3660 1960 1101 3504 1077 3405 2608 1773 2370 2059 1764 1441 1711 3013 3587 2715 64 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7693 8735 894 1115 358 2187 8914 8488 65(L) 891 2044 4713 4184 1318 4146 3223 1658 3880 2504 198 3858 3970 3332 3681 3346 2320 337 2643 2499 65 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8484 66(R) 3030 3584 3982 2344 4551 3458 1269 3800 2924 3397 2757 2153 3376 852 3034 2903 2656 3598 3188 3160 66 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8480 67(F) 2326 1891 4869 4440 2685 4485 3701 2340 4251 921 863 4124 4292 3931 4198 3737 2296 2290 3181 2708 67 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8476 68(R) 3054 3592 3980 2373 4560 3469 1297 3823 1781 3421 2785 2179 3395 881 3702 2930 2684 3621 3206 3181 68 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8472 69(F) 3145 2644 4628 4516 2787 4252 1269 1824 4116 1928 1754 3303 4186 3281 3729 3432 3069 2671 580 2537 69 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8468 70(V) 2450 1960 5111 4795 2565 4880 4855 1914 4732 1343 1286 4581 4668 4656 4868 4252 2448 3379 4256 3756 70 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8464 71(N) 1525 2287 1522 1828 4053 2153 2507 4149 2667 4251 3455 3942 2849 2311 3012 1045 1898 3178 4209 3651 71 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8459 72(V) 2838 2616 4834 4880 3213 3955 4416 610 4757 2310 2253 4383 4345 4643 4642 3732 3007 3763 4157 3886 72 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8455 73(V) 2243 1824 4833 4454 2402 4433 4113 1762 4305 1350 1201 4152 4338 4160 4358 3722 1258 3104 3760 3307 73 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8451 74(D) 2668 4634 3697 1529 4809 2393 1860 4835 2286 4663 4116 996 2985 1560 3281 2276 2774 4274 4791 3728 74 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8447 75(L) 3647 3060 5937 5371 1118 5682 4333 610 5087 3001 2533 5429 4784 3925 4592 5073 3493 1435 2916 3081 75 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8443 76(M) 1722 1497 3954 3355 1014 848 2243 1549 2997 978 3830 2966 3285 2560 2824 2460 1661 398 1947 1696 76 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8439 77(K) 1675 2604 1914 1147 3023 2553 912 2542 2655 2539 1775 1279 2595 506 1666 1628 1008 985 2674 2295 77 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8435 78(L) 4119 3542 5389 5357 2027 4767 4358 1609 5118 3293 979 5230 4771 4474 4724 5069 4106 2331 3412 3400 78 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8430 79(Q) 1357 2106 1510 1196 2392 2243 1288 2153 824 371 1573 1327 2535 3535 1081 1033 1413 1913 2627 2087 79 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8426 80(P) 1393 2284 1577 1362 3557 2164 1558 3266 1432 3269 2444 1443 2774 1192 1186 2003 1560 2662 3389 2924 80 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8422 81(K) 1010 2349 767 1764 2657 1927 589 2362 1961 301 1468 584 2042 153 523 917 946 1969 2533 1906 81 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8418 82(H) 956 2424 615 1087 2749 1853 2292 2494 2040 2432 1522 1288 1978 92 585 753 897 2050 2592 1916 82 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 1873 7946 474 894 1115 701 1378 8914 8414 83(G) 853 1186 1419 1603 2565 3376 1668 2627 1844 2759 2200 1431 1859 1709 1902 1054 1192 2034 2282 2387 83 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 36 6109 7151 894 1115 2826 219 8914 8409 84(A) 2862 488 1346 1349 1807 785 1257 1119 1298 1627 1088 934 1428 1183 1403 201 309 714 2107 1798 84 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 36 6109 7151 894 1115 2826 219 8914 8405 85(L) 1446 1261 2639 2396 166 2529 1687 443 1969 2652 716 2264 2607 1810 1925 2083 1460 71 1338 963 85 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 36 6109 7151 894 1115 431 1952 8914 8401 86(E) 1620 3077 205 2781 2713 2016 1027 3107 935 3054 2295 1905 2376 703 1502 1416 1618 2704 2909 1972 86 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7696 8738 894 1115 1502 628 8914 8397 87(H) 1687 3020 524 508 3027 2149 4043 3137 478 3027 2263 1907 2446 2055 794 1508 1649 2742 2951 2237 87 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7696 8738 894 1115 360 2179 8914 8392 88(P) 2450 4345 1816 1516 4562 2311 1714 4514 2022 4375 3745 914 3318 1394 2920 2098 2536 3971 4556 3521 88 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8388 89(H) 2178 3297 1451 1311 3729 2695 4349 3419 165 3210 2472 1349 2852 755 1705 2018 2038 3087 3149 2733 89 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8384 90(G) 1135 2508 960 1390 3988 2880 1982 3774 2052 3851 3054 1368 2715 1683 2580 1591 1835 3036 4050 3424 90 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8379 91(M) 3620 3042 5906 5341 1118 5639 4298 608 5051 2656 3645 5383 4766 3909 4566 5021 3469 1425 2906 3062 91 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8375 92(S) 1385 2104 1840 2115 4039 2094 2642 4072 2802 4207 3375 1808 2809 2466 3073 3255 1780 3055 4205 3726 92 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8371 93(D) 2954 4081 3777 1338 4930 2812 2481 5106 2896 4959 4423 1697 1695 2236 3689 2728 3134 4489 4667 4137 93 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8366 94(R) 847 2632 1132 2003 3046 2175 695 2716 1321 2604 1741 819 2258 250 2101 1179 1206 2313 2696 2152 94 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8362 95(D) 2629 4604 3009 2799 4770 2374 1828 4782 2234 4614 4051 973 2960 1523 3216 2241 2730 4224 4757 3689 95 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8358 96(F) 4587 3562 5019 5350 4191 4887 1144 3464 4916 2789 2878 3547 4760 3678 4301 4156 4453 3634 393 2478 96 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8353 97(D) 988 3875 2907 1483 4100 2208 1447 3972 1539 3865 3114 1567 2675 1089 2294 1800 2126 3467 4055 3126 97 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8349 98(S) 1458 2457 1399 292 2897 1890 750 2648 1311 2615 1722 599 2098 317 922 1668 1053 2187 2800 2113 98 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8344 99(L) 3562 3022 4998 4839 384 4758 2089 1358 4382 2908 731 3989 4502 3590 4015 4103 3467 1964 1321 2369 99 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8340 100(F) 4720 3927 5139 5454 4513 4504 2520 3938 5326 3307 3429 4469 4782 4575 4818 4787 4810 4172 1812 736 100 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8335 101(T) 1345 1936 3485 3748 4307 1297 3469 4115 3845 4377 3519 2741 2943 3484 3725 1588 3638 3022 4411 4351 101 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8331 102(P) 1040 1933 1444 985 2825 1942 1172 2498 1349 2593 1749 1119 2171 794 1131 1051 1955 2025 2847 2320 102 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8326 103(D 2253 3538 3694 855 4532 2334 2017 4570 2397 4510 3869 1194 2951 1736 3233 1140 2471 3857 4578 3695 103 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 1873 7946 474 894 1115 701 1378 8914 8322 104(G 853 1186 1419 1603 2565 3376 1668 2627 1844 2759 2200 1431 1859 1709 1902 1054 1192 2034 2282 2387 104 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 36 6109 7151 894 1115 133 3504 8914 8317 105(K 2513 3301 3024 1871 4114 3126 1139 3484 3142 3184 2485 1825 3101 717 1860 2405 1288 3220 3066 2911 105 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8313 106(P) 3125 3472 2885 3107 3559 3370 2901 4916 3129 4732 4256 3117 3925 3250 3218 3230 3395 4360 3686 3131 106 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8308 107(V 2449 1961 5107 4792 2559 4869 4842 1787 4726 1338 1282 4575 4662 4648 4859 4241 2447 3416 4245 3747 107 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8304 108(I) 2038 1784 4073 3538 1026 3659 2506 3064 3042 1092 254 3185 3570 2750 2918 2809 1976 421 2001 1514 108 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7946 8988 894 1115 701 1378 8914 8299 109(F) 4720 3927 5139 5454 4513 4504 2520 3938 5326 3307 3429 4469 4782 4575 4818 4787 4810 4172 1812 736 109 149 500 233 43 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249 14 7947 8989 894 1115 701 1378 8914 8234 123(Y 4607 3570 5022 5359 2289 4894 1139 3493 4923 2816 2907 3546 4765 3679 4305 4161 4470 3658 387 4501 123 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8229 124(D 1348 2787 2242 415 3129 2090 2199 2863 263 2769 1903 717 2264 1787 2067 1197 1284 2435 2889 2250 124 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8224 125(R 4488 4181 4789 4318 5193 4148 3436 5568 2393 5152 4748 4156 4479 3286 4202 4614 4467 5273 4267 4649 125 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8219 126(T) 900 1750 1012 977 1985 754 770 1034 516 1791 1012 790 1172 435 956 941 1779 1338 2179 1644 126 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 510 7947 1783 894 1115 701 1378 8914 8214 127(N 1735 2957 337 503 3341 2022 1191 3403 833 3305 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720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8190 132(H 1265 1465 2433 2035 1445 2449 3775 1372 1614 1394 839 1901 2701 1628 1732 1623 2191 807 1952 1432 132 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8185 133(V 2378 2013 4879 4522 2020 4336 3971 321 4271 861 2587 4174 4288 3996 4221 3659 2396 3344 3447 3110 133 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8180 134(H 1298 2431 1191 1210 2723 2201 3201 2373 52 2378 1563 884 2290 333 2092 1222 1207 912 2555 2032 134 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8175 135(G 4088 3924 4774 5139 5615 3825 4753 6303 5453 6014 5662 4812 4539 5232 5106 4370 4472 5527 4696 5561 135 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8170 136(Y 4791 3935 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140 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8145 141(T) 1448 2786 1566 494 3406 2031 1186 3187 1023 3164 2316 1513 2396 794 1603 1061 2567 2675 3364 2628 141 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8140 142(I) 1880 1845 4196 3993 2337 3273 3434 2845 3687 1437 1287 3407 3652 3527 3649 2612 2766 19 3410 3029 142 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8134 143(T) 1427 2097 2121 2355 3933 2165 2713 3790 2759 4026 3246 1603 2869 2560 2943 1604 3622 2931 4094 3692 143 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8129 144(T) 2430 2823 4179 4462 4591 3027 4037 4454 4406 4707 4191 3664 3724 4237 4215 2689 4010 3747 4431 4588 144 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8124 145(P) 1505 2080 3392 3633 4254 2301 3425 4043 3658 4296 3530 2794 3850 3427 3585 1744 1558 3083 4340 4254 145 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8119 146(F) 3994 3227 4915 5043 3832 4698 1205 2713 4616 2132 2459 3492 4563 3530 4094 3916 3878 2950 467 2613 146 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8114 147(D 2625 4424 3697 649 4717 2403 1898 4810 2318 4653 4097 1859 2997 1605 3282 2269 2750 4226 4723 3698 147 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8109 148(M 3455 2954 5636 5138 1157 5278 4061 621 4736 1276 4790 5054 4653 3832 4350 4674 3346 1354 2859 2873 148 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8103 149(M 737 1039 2128 1551 1071 2251 1108 553 1284 925 2340 1505 2347 1148 1957 1292 992 1190 1511 1125 149 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8098 150(V 1589 1340 3842 3261 1369 3310 2277 1533 2856 783 2491 2892 3292 2604 1266 2434 1540 2552 2138 1796 150 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8093 151(C 948 2699 2923 2314 734 2453 1287 272 1969 629 6 1970 2503 2182 1898 1524 890 1281 1253 1875 151 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8087 152(N 1513 2274 1534 1835 4051 2147 2506 4137 2664 4241 3442 3900 2843 2309 3007 1223 1886 3166 4207 3653 152 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8082 153(Q 1078 2597 1398 1298 2904 570 660 2670 1137 2607 1702 1544 2056 1583 856 929 1031 2214 2776 2059 153 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8077 154(M 2272 1957 4590 3986 1077 4004 2865 1417 3652 2191 2665 3646 3782 3053 3407 3149 1251 644 2313 2179 154 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8071 155(D 2253 3537 3692 855 4532 2333 2017 4569 2396 4510 3868 1194 2951 1735 3232 1151 2470 3856 4577 3695 155 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8066 156(R 4488 4181 4789 4318 5193 4148 3436 5568 2393 5152 4748 4156 4479 3286 4202 4614 4467 5273 4267 4649 156 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8061 157(F) 4608 3568 5026 5363 3806 4898 1136 3489 4926 2811 2901 3545 4766 3678 4306 4161 4469 3654 384 3465 157 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8055 158(H 1044 2139 1080 861 3017 1958 3956 2774 984 2841 1992 1421 2333 887 1434 909 1252 2260 3059 2457 158 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8050 159(L) 1995 1713 4228 3647 1208 3730 2645 1439 3309 2122 164 3321 3602 1641 3161 2864 1930 1443 2284 2058 159 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8044 160(A 2383 1556 2401 2072 2208 2210 1881 1266 1832 1939 1334 1898 2639 1793 2020 1435 1318 1882 2638 2245 160 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8039 161(I) 855 2283 1296 661 2565 2186 701 1827 1284 2220 1411 884 2249 1555 1499 1169 1126 1886 2436 1937 161 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8033 162(D 1082 2449 2531 267 2754 1915 698 2479 1332 2474 2209 579 2090 271 827 743 1039 2075 2684 2016 162 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8028 163(V 2572 1717 4125 4110 2971 2517 3473 716 3875 2280 1954 3032 3163 3569 3746 1871 1754 2604 3798 3481 163 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8022 164(M 2861 2402 5299 4748 1283 4880 3804 1499 4501 2317 2674 4552 4398 3712 4200 4119 2770 1228 2863 2864 164 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8017 165(Q 2277 4157 2369 2212 4337 2257 1565 4246 1745 4116 3418 845 2768 2820 2553 1949 2331 3727 4303 3315 165 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 1378 8914 8011 166(W 895 1130 1817 1235 1105 2196 2271 700 958 627 314 1121 2263 882 1030 1203 832 1095 2753 1082 166 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 14 7947 8989 894 1115 701 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894 1115 2557 269 8914 7954 176(N 707 1550 295 469 2833 2212 1124 2747 1097 2834 2065 2825 1857 854 1516 754 965 2073 2881 2341 176 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 686 6760 1453 894 1115 2557 269 8914 7948 177(L) 1446 1261 2639 2396 166 2529 1687 443 1969 2652 716 2264 2607 1810 1925 2083 1460 71 1338 963 177 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 37 6111 7153 894 1115 435 1941 8914 7943 178(C 1406 3275 2556 2171 2790 2067 1965 2431 1828 2675 1880 1777 2338 1781 1478 1008 1091 1870 3029 2681 178 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7695 8737 894 1115 1509 624 8914 7937 179(N 1616 1783 884 388 2121 1793 600 1720 1446 1882 1075 1636 1951 220 618 795 775 841 2206 1651 179 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7695 8737 894 1115 358 2187 8914 7931 180(H 826 2030 1249 286 2281 1850 2945 1963 256 2046 2137 570 1980 190 741 713 829 1630 2333 1721 180 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7948 8990 894 1115 701 1378 8914 7925 181(L) 1888 1658 4011 3398 1778 3497 2279 1355 1112 2128 19 3057 3387 2573 2834 2608 1812 657 1938 1655 181 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7948 8990 894 1115 701 1378 8914 7919 182(C 1582 2847 3677 3112 1572 3286 2308 2397 1781 1057 639 2820 3303 2550 2691 2415 1546 1459 2297 1922 182 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7948 8990 894 1115 701 1378 8914 7913 183(Q 889 2895 1501 1223 3185 1975 867 2968 610 2899 2019 1640 2225 2395 1182 1154 1298 2505 3073 2320 183 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7948 8990 894 1115 701 1378 8914 7907 184(M 624 2075 807 1189 2318 1853 499 2000 1304 56 1892 530 1940 75 1281 778 783 1655 2297 1689 184 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 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3117 1691 1757 2379 2109 2248 2689 2425 189 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7948 8990 894 1115 701 1378 8914 7870 190(V 1079 1178 2219 953 1217 2479 1279 1243 1083 512 362 1650 2531 1279 1608 1507 1021 1902 1713 1318 190 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7948 8990 894 1115 701 1378 8914 7864 191(E) 1354 2910 1356 2455 3203 1991 882 2982 1322 2913 2038 608 2244 460 1170 1178 1230 2523 3086 2339 191 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7948 8990 894 1115 701 1378 8914 7858 192(H 995 2611 2882 2350 1156 2330 3755 1388 2017 1028 400 2003 2551 1787 2019 1469 1023 451 1652 1275 192 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7948 8990 894 1115 701 1378 8914 7852 193(F) 873 2098 751 1196 2231 1853 534 2026 137 2078 1223 1242 1958 115 1091 800 953 1680 2329 1708 193 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7948 8990 894 1115 701 1378 8914 7845 194(Q 684 2845 419 1349 3160 1991 875 2930 582 2870 1997 2370 2238 2528 1121 1166 1300 2477 3045 2315 194 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7948 8990 894 1115 701 1378 8914 7839 195(Y 3797 3103 4834 4924 2259 4579 1182 2596 4505 2328 2267 3426 4481 3479 4016 3789 3700 1231 456 4130 195 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7948 8990 894 1115 701 1378 8914 7833 196(I) 1842 1480 3746 3421 2131 2613 2685 2759 3142 1622 1194 2759 3062 2873 3093 1882 1362 243 2830 2485 196 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7948 8990 894 1115 701 1378 8914 7827 197(C 917 2447 2927 2307 742 2421 1266 267 1915 627 2390 1950 2475 1643 1928 1495 858 1278 1243 1688 197 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7948 8990 894 1115 701 1378 8914 7820 198(E) 2062 3846 1749 2650 4068 2200 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210 466 720 275 394 45 96 359 117 369 294 249 15 7948 8990 894 1115 701 1378 8914 7788 203(M 2141 1848 4423 3825 1086 3850 2726 2359 3484 1059 3290 3479 1362 2946 3270 2988 2066 538 2251 2080 203 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7948 8990 894 1115 701 1378 8914 7781 204(P) 4497 4117 4899 5251 5560 4139 4798 6328 5454 5976 5741 5026 4302 5328 5104 4798 4844 5739 4665 5484 204 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7948 8990 894 1115 701 1378 8914 7775 205(E) 2636 4572 1810 3431 4760 2390 1841 4772 2235 4607 4040 995 2973 1537 3199 2253 2735 4218 4733 3691 205 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7948 8990 894 1115 701 1378 8914 7768 206(I) 2944 2476 5069 4784 2837 4680 2731 3180 4519 584 703 4172 4449 3817 4249 3998 2901 494 1967 1074 206 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7948 8991 894 1115 701 1378 8914 7762 207(T) 1149 1001 3245 2651 830 2680 1537 200 2295 678 115 2252 2729 1967 2187 1772 2521 1202 1425 1941 207 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7948 8991 894 1115 701 1378 8914 7755 208(N 968 3781 2422 1475 4014 2190 1399 3875 1453 3773 3006 2431 2639 1035 2184 1745 2052 3376 3962 3054 208 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7948 8991 894 1115 701 1378 8914 7748 209(W 1214 2627 4165 4274 2248 2999 2977 3589 3845 3575 3208 3402 3626 3726 3688 2549 2659 3197 5855 1919 209 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7948 8991 894 1115 701 1378 8914 7741 210(V 1358 2495 1397 737 2853 2276 735 2472 1340 2436 1616 951 1242 1587 1414 1276 1250 1808 2580 2097 210 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7949 8991 894 1115 701 1378 8914 7735 211(W 3498 2927 4676 4658 236 4377 1183 2501 4186 2296 2147 3324 4322 3336 3785 3580 3418 1241 5352 2614 211 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 16 7949 8991 894 1115 701 1378 8914 7728 212(H 955 1646 1373 926 1547 635 2162 1632 859 1839 1087 1075 2103 763 1254 967 989 1372 2122 1543 212 * * * * * * * * * * * * * * * * * * * * * * * * * * * 8914 0
(332) TABLE-US-00019 TABLE 14 HMMER2.0 [2.2g] NAME PTA_PTB_exp_seqs LENG 355 ALPH Amino RF no CS no MAP yes COM /app/public/hmmer/current/bin/hmmbuild PTA_exp_hmm PTA_PTB_exp_seqs.aln COM /app/public/hmmer/current/bin/hmmcalibrate PTA_exp_hmm NSEQ 10 DATE Mon Nov 30 18:53:24 2009 CKSUM 2317 XT 8455 4 1000 1000 8455 4 8455 4 NULT 4 8455 NULE 595 1558 85 338 294 453 1158 197 249 902 1085 142 21 313 45 531 201 384 1998 644 EVD 321.869965 0.133020 A C D E F G H I K HMM m->m m->i m->d i->m i->i d->m d->d b->m m->e L M N P Q R S T V W Y 438 * 1933 1(M) 1511 1258 3275 2807 2269 2995 1346 266 2474 800 3792 2446 2858 1968 2335 2172 1449 138 793 87 26 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 23 6577 7619 894 1115 701 1378 438 * 2(M) 426 1446 3780 3363 1521 2856 2514 90 3024 729 4481 2828 3133 2721 2939 2091 1564 1100 2424 2124 27 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 10 7762 8804 894 1115 701 1378 * * 3(D) 898 2223 2613 202 2512 1832 557 2225 779 2241 1722 504 1962 125 678 389 566 1836 2468 1816 28 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 4(L) 3343 2811 5687 5115 2061 5345 3911 1769 4867 2527 56 5003 4615 3800 4424 4629 3207 1173 2733 2675 29 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 5(F) 3039 2583 5400 4800 2731 4939 3665 2351 4517 1482 1999 4610 4383 3595 4132 4161 2919 1092 2663 2673 30 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 6(E) 1913 3483 329 3240 3847 2201 1343 3669 1205 3580 2799 844 2614 1375 1770 328 1937 3187 3742 2936 31 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 7(K) 892 2327 754 192 2667 1850 472 2397 1835 2332 1421 831 1938 1186 1037 1297 497 1961 2483 1835 32 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 8(I) 514 1727 4577 4078 1745 4080 3252 2588 3824 1712 655 3731 3953 3476 3724 3273 2060 1203 2879 2568 33 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 9(W) 681 1696 954 400 531 1888 578 324 1375 1628 839 651 1974 1030 909 344 741 1214 2014 1462 34 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 10(E) 1665 3313 1727 2676 3578 225 1130 3392 1004 3309 2472 694 2435 928 1638 306 1659 2915 3488 2672 35 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 11(K) 1580 1884 1764 1020 2907 2481 845 2479 1956 772 1679 1179 2514 1923 1777 1525 1447 2203 2604 2197 36 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 12(A) 2315 1624 4424 3936 1894 3936 3165 1771 3688 61 834 3585 3870 3418 3634 3127 1946 1865 2913 2538 37 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 13(K) 240 2148 1205 600 2451 2088 695 2066 2741 2143 1331 829 2179 293 688 1066 326 519 2394 1872 38 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 14(Q) 251 2245 585 74 2564 1048 408 2314 875 2260 1335 384 515 1120 955 394 715 1866 2429 1747 39 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 15(D) 511 2241 1741 132 2549 1792 445 2281 1286 455 1337 435 1886 971 685 711 760 1857 2426 1765 40 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 16(K) 884 2341 1633 125 2653 223 507 2400 1752 2356 1445 919 707 57 629 761 829 80 2535 1850 41 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 17(K) 535 1741 1193 620 1889 2043 709 327 2612 1654 903 840 2126 854 679 1007 888 295 2069 1566 42 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 18(K) 2416 3238 2867 1785 4015 3059 1120 3414 2580 3139 2429 1764 3047 698 2089 2312 2234 3140 3041 2859 43 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 19(I) 2521 2024 5153 4783 2223 4929 4595 3447 4687 294 972 4589 4629 4430 4724 4267 2498 1528 3873 3540 44 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 20(V) 213 1847 4873 4520 2465 4420 4247 1419 4388 1381 1247 4198 4362 4252 4452 3727 2270 3336 3880 3419 45 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 21(F) 3693 3087 5586 5265 3261 5304 2888 912 4973 2381 226 4694 4708 3838 4472 4669 3558 1678 1934 1153 46 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 22(P) 1357 1977 3526 3796 4305 2211 3504 4098 3878 4361 3542 2788 3823 3529 3749 1636 1847 3043 4411 4353 47 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 23(E) 2645 4564 1495 3508 4762 2401 1853 4778 2245 4613 4046 1009 2982 1550 3202 2265 2745 4225 4728 3699 48 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 24(G) 828 1562 2312 1921 2620 2562 1856 2263 1813 2508 1731 1730 2452 1660 2098 554 782 1818 2902 1503 49 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 25(E) 1027 2214 837 2256 2461 1966 641 2117 974 634 1343 652 2082 229 588 955 1152 1790 2438 1847 50 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 26(W) 150 757 49 299 277 651 839 485 392 910 599 300 401 623 162 99 38 448 1107 126 51 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 27(D) 2650 4626 3535 2005 4794 2383 1844 4814 2264 4643 4090 983 2972 1542 3255 2259 2754 4254 4780 3711 52 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 28(E) 884 1962 861 2360 2149 1901 576 272 764 268 1086 611 909 182 632 849 822 1489 2226 1648 53 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 29(R) 4488 4181 4789 4318 5193 4148 3436 5568 2393 5152 4748 4156 4479 3286 4202 4614 4467 5273 4267 4649 54 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 30(V) 1615 1406 3692 3190 1680 3315 2460 2105 2933 1136 730 1527 3366 2700 2939 2463 576 2489 2439 2054 55 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 31(L) 2893 2389 5409 4931 1519 5116 4253 2238 4753 2516 300 4794 4601 4030 4524 4423 2823 819 3207 3187 56 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 32(K) 810 2256 666 736 2577 491 428 19 1965 2266 1348 419 1872 1176 702 323 747 1879 2434 1767 57 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 33(A) 3609 2508 4184 4493 4592 2737 3989 4421 4496 4701 4051 3440 3474 4171 4253 2294 2487 3536 4484 4643 58 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 34(A) 2937 1528 3502 3308 3125 1952 2769 2630 3081 3039 2271 2390 2643 2779 3078 549 1011 493 3475 3206 59 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 35(H) 804 2234 624 1374 2527 1779 1863 180 65 2239 1335 424 1886 14 572 1383 764 1842 2435 1765 60 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 36(K) 927 1951 1003 613 2143 1960 589 446 1801 1893 1449 680 2039 201 1687 903 856 1504 2198 1303 61 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 37(I) 3179 2647 5611 5124 1356 5341 4317 2901 4907 2264 148 5052 4698 4019 4582 4697 3088 546 3099 3137 62 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 38(I) 1305 916 2346 1758 894 2325 1140 1372 98 911 104 1638 2383 1135 1607 1364 839 747 1366 994 63 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 39(K) 103 2280 655 781 2607 33 431 2352 2090 2292 1371 817 1875 1393 703 697 754 1906 2454 1782 64 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 40(D) 876 2355 1990 1398 2673 1797 489 2425 861 2368 1450 862 1916 35 739 749 818 1976 2535 1245 65 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 41(G) 1910 3521 1036 459 3833 2939 1305 3641 669 3546 2760 819 2598 922 1710 1672 1918 3170 3710 2907 66 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 42(L) 2752 2316 5181 4626 1316 4723 3629 2215 4364 2228 104 4388 4302 3622 4077 3942 2665 822 2775 2716 67 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 43(A) 2595 1592 4065 3655 1874 3190 2875 69 3364 237 870 3140 3438 3084 3305 2430 1771 2003 2788 2442 68 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 44(K) 781 2248 1290 1182 2566 1751 412 18 1303 2262 1338 836 1846 47 956 662 674 1869 2432 1751 69 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 45(P) 1999 1685 4334 3818 1612 3852 2976 1828 3535 394 557 3486 3050 3199 3450 3026 1964 1176 2676 2369 70 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 46(V) 2452 1958 5119 4802 2578 4903 4884 2288 4745 1352 1293 4595 4680 4675 4887 4275 2449 3237 4279 3776 71 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 47(L) 2998 2538 5403 4947 1395 4989 4115 173 4693 2765 209 4769 4551 3928 4414 4328 2940 1827 3062 3033 72 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 48(V) 2510 2021 5126 4737 2130 4872 4437 1947 4622 1286 893 4529 4574 4319 4624 4190 2483 2866 3732 3431 73 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 49(G) 931 1908 3564 3879 4400 3463 3561 4237 4052 4490 3601 2764 2927 3600 3877 1557 1777 3055 4501 4478 74 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 50(N) 91 2861 975 587 3637 2069 1361 3439 1272 3412 2579 2972 2496 985 1877 1896 1654 2876 3606 2851 75 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 51(E) 1387 2840 463 2722 3195 2028 944 2952 645 2907 2050 684 1763 936 1162 1231 313 2508 3091 2377 76 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 52(N) 1378 2248 457 624 806 1762 434 2299 865 2258 1342 1494 1863 20 534 341 740 1863 2438 1760 77 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 53(E) 975 2349 734 2085 2655 1807 516 2399 1272 2359 1451 445 1934 67 640 772 840 10 2541 1856 78 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 54(I) 2526 2030 5153 4776 2181 4921 4549 3338 4675 616 934 4580 4616 4394 4697 4253 2501 1644 3821 3507 79 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 55(K) 398 2606 1173 479 3022 2165 660 2692 2540 2573 1705 805 2232 1750 1236 1146 1168 2286 2659 2117 80 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 56(E) 1257 2318 622 1753 2641 1799 475 2386 1275 2333 1417 437 1908 1398 542 391 801 1943 2501 1824 81 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 57(F) 471 982 1994 1417 1160 2213 1003 523 1101 784 172 584 2279 1043 1416 1230 532 1019 1408 1017 82 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 58(A) 3609 2508 4184 4493 4592 2737 3989 4421 4496 4701 4051 3440 3474 4171 4253 2294 2487 3536 4484 4643 83 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 59(K) 252 2225 594 101 2531 1206 438 2270 1451 150 1326 673 1865 14 536 340 739 1841 2425 1752 84 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 299 7942 2449 894 1115 701 1378 * * 60(E) 1167 2286 732 1831 2598 1667 407 2354 951 2301 1391 300 1816 40 560 783 749 1906 2475 1775 85 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7654 8696 894 1115 1585 585 * * 61(H) 815 727 2621 2031 1145 2289 2094 200 1722 1681 107 494 2340 1435 1697 1354 755 1054 1030 564 86 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 11 7654 8696 894 1115 337 2262 * * 62(N) 1704 3281 1777 397 3622 1646 1188 3429 1036 3352 2526 2598 2480 796 1047 1495 1710 2949 3526 2731 87 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 63(M) 20 1214 3805 3190 774 3078 1973 1042 2815 1458 2157 2723 3056 2395 2615 2183 1382 1944 1731 1436 88 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 64(D) 605 2467 2563 401 3040 1918 912 2798 624 2777 1892 929 2187 492 1157 1186 553 2307 2970 2279 89 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 65(L) 3447 2885 5829 5243 1145 5544 4253 1795 5014 2666 2377 5248 4707 3887 4549 4864 3303 1162 2916 3088 90 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 66(D) 250 2410 1559 1458 2748 39 576 2502 207 2454 1544 475 1976 130 729 1014 986 2052 2631 1936 91 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 67(G) 2653 3216 2503 2570 4722 3367 2672 4647 1672 4521 3864 2587 3560 2414 2013 2728 2874 3999 4152 4117 92 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 68(V) 1628 1694 4600 4141 2065 4161 3476 2051 3925 24 959 3809 4058 3687 3896 3377 2060 2431 3181 2784 93 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 69(Q) 1107 2491 869 2104 2843 2005 610 2548 1501 2474 1449 645 2106 2280 390 986 1031 2135 2613 2004 94 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 70(I) 2462 1971 5120 4803 2532 4890 4848 3484 4737 1301 1256 4591 4671 4645 4865 4262 2459 1950 4227 3745 95 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 71(V) 1021 845 3291 115 783 2586 1463 1772 2289 711 1731 2204 2623 1926 2122 1672 963 1810 1336 1149 96 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 72(D) 4232 4465 4158 2649 5412 3676 3575 6036 4125 5722 5419 2993 4216 3512 4617 4077 4401 5553 4696 4914 97 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 73(P) 1143 1675 3005 2989 3065 2064 2700 565 2869 2950 2300 2338 3698 2683 2908 587 1504 2001 3467 3104 98 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 74(H) 1253 2202 717 1654 2478 1838 2550 2187 868 372 1310 496 1938 63 532 776 809 1802 2405 1767 99 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 75(T) 1143 1868 2054 2081 3765 1965 2374 3533 2354 3699 2834 2518 2626 2132 2643 882 2968 2663 3905 3493 100 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 76(Y) 1148 1566 1831 1449 1231 2241 1207 1418 1361 1676 1002 1156 2489 1227 1649 1580 1191 429 1711 3317 101 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 77(W) 150 757 49 299 277 651 839 485 392 910 599 300 401 623 162 99 38 448 1107 126 102 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 78(E) 1130 2548 993 1837 2823 1914 731 2562 425 645 1679 557 1649 306 961 330 1088 2151 2765 2070 103 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 79(K) 705 2293 914 357 2734 819 598 2445 2189 2399 1507 628 2037 153 975 617 923 2013 2558 1945 104 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 1567 7942 603 894 1115 701 1378 * * 80(E) 1455 2350 50 3362 2809 1547 927 2700 845 2735 2178 487 2000 660 1278 1311 1525 2365 2625 2220 105 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 26 6400 7442 894 1115 1808 485 * * 81(M) 551 928 1713 589 942 1741 615 5 559 521 2643 741 1867 429 905 804 519 1266 1450 997 106 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 19 6823 7865 894 1115 167 3194 * * 82(K) 936 1587 1304 718 1678 2068 727 1273 1622 91 1465 909 2141 440 1553 1037 1209 1083 1910 1220 107 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 83(Q) 216 3288 1080 1963 3557 2088 1124 3366 985 3287 2450 697 2432 3156 1608 1439 1648 2893 3467 2658 108 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 84(E) 661 2268 1307 1400 2586 40 425 2337 12 2283 1359 392 1857 1107 956 675 492 1889 2452 1768 109 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 85(M) 2903 2473 4675 4372 1750 4229 1624 1350 3989 445 3626 3447 4067 3192 3628 3390 2808 1716 937 3375 110 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 86(A) 1901 908 3172 2568 899 2621 1244 994 2224 146 105 2185 2665 1906 2124 1705 1008 1900 1447 1095 111 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 87(Q) 1463 3014 2011 328 3316 2053 949 3092 641 3010 2150 2270 2319 2525 696 1279 1430 2636 3169 2433 112 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 88(A) 2248 2123 670 520 2701 1892 856 2400 496 2451 1589 769 2126 446 1017 948 1463 1971 2692 2080 113 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 89(F) 2727 2338 4825 4352 3286 4296 2265 816 3996 1550 226 3736 4033 3222 3652 3458 2628 1306 3005 774 114 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 90(Y) 2126 1823 4287 3802 723 3742 2094 327 3430 1159 480 3251 3663 2945 3227 2891 2068 2283 1586 3201 115 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 91(E) 703 3188 772 3501 4265 2379 2017 4040 2108 4114 3438 1356 2948 1721 2684 2006 2303 3449 4241 3565 116 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 92(R) 2413 2234 4171 3369 1308 3884 2460 877 2018 2018 224 3190 3714 2356 2764 3076 2310 961 2462 2283 117 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 93(R) 738 2612 3347 2738 4144 2736 2052 3768 900 3705 3029 2409 3189 1725 3817 2171 2228 3209 3627 3498 118 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 94(K) 503 2445 1015 703 3184 562 1018 2893 2978 2852 1992 704 2331 602 835 1206 1297 2406 2999 2416 119 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 95(H) 928 2405 563 997 2720 1543 2401 2472 959 2417 1504 868 1955 84 643 392 872 2026 2586 1897 120 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 96(K) 4125 4091 4017 3615 5101 3960 2958 5182 3972 4825 4345 3552 4249 2709 1809 4154 4044 4895 4132 4389 121 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 809 7942 1234 894 1115 701 1378 * * 97(G) 2666 2751 3377 3700 4393 3753 3528 4841 4031 4741 4286 3394 3411 3824 3848 2912 3053 4080 3719 4290 122 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7148 8190 894 1115 541 1677 * * 98(M) 412 754 2560 1966 741 2291 1150 981 1684 598 3179 1245 2352 1421 1700 1351 649 877 1235 874 123 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 10 7762 8804 894 1115 413 2007 * * 99(T) 2430 2823 4179 4462 4591 3027 4037 4454 4406 4707 4191 3664 3724 4237 4215 2689 4010 3747 4431 4588 124 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 100(P) 270 1065 2103 243 1059 2317 1151 1403 1372 790 240 1539 2257 1198 1557 1358 904 375 1531 1145 125 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 101(E) 2392 4275 1163 3312 4486 2295 1667 4423 1934 4288 3633 894 1125 1341 2802 2049 2468 3888 4470 3454 126 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 102(Q 297 2471 1012 160 2788 1849 577 2543 1870 2482 1573 944 1994 2462 693 848 930 2093 2649 1955 127 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 103(A 3227 1594 3807 3849 3646 1945 3198 3067 3621 3576 2783 2572 2704 3241 3481 576 1455 241 3977 3779 128 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 104(D 1261 2267 1791 738 2557 1813 517 2280 130 2273 1380 464 1934 76 1383 771 825 386 2479 1813 129 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 105(K 415 2546 735 1917 2896 1973 635 2621 2122 2543 1653 608 2101 1705 517 981 1047 2189 2683 2042 130 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 106(M 308 827 2563 1964 795 2335 1167 1419 607 677 2765 1747 2393 1425 1700 1388 1409 241 2166 912 131 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 107(M 2838 2385 5273 4720 1280 4846 3769 1119 4469 2241 2938 4517 4377 3690 4172 4080 2748 1487 2847 2840 132 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 108(K 1031 2407 918 1080 2761 1972 550 2464 1949 434 1505 616 2053 100 1475 1180 952 2050 2533 1929 133 149 500 233 43 381 400 106 626 210 466 720 275 394 45 96 359 117 369 294 249 169 3204 8984 206 2913 701 1378 * * 109(D 4232 4465 4158 2649 5412 3676 3575 6036 4125 5722 5419 2993 4216 3512 4617 4077 4401 5553 4696 4914 135 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 110(P) 893 2081 1781 940 1121 2183 978 444 1070 1013 334 1295 2567 933 1326 1202 842 568 1538 1138 136 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 111(N 1006 916 2854 2266 891 2503 1385 865 1966 1747 97 1859 2563 1688 1947 70 954 854 1419 1063 137 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 112(Y 4083 3282 4926 5082 1832 4728 1188 2828 4652 2243 2245 3495 4595 3549 4122 3951 3965 3052 449 4351 138 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 113(F) 4576 3559 5015 5343 4263 4881 1152 3449 4910 2776 2865 3550 4757 3679 4300 4154 4444 3622 401 2124 139 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 114(G 2612 1827 3654 3943 4336 2661 3533 4149 4027 4411 3508 2727 2864 3570 3838 1474 1691 2967 4475 4432 140 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 115(T) 1598 1522 3726 3555 2787 2149 2868 1443 3288 2470 1892 2585 2799 2974 3213 1463 2927 1522 3313 3022 141 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 116(M 845 1514 3507 3305 2427 2152 2615 1675 2968 2025 4655 2481 2760 2728 2908 1457 695 1444 2989 2699 142 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 117(M 103 2295 4810 4261 1123 4222 3233 505 3905 1982 4088 3963 4004 3263 3656 3428 2527 1018 2542 2485 143 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 118(V 249 1764 4559 4171 2124 3892 3557 400 3936 216 1010 3749 3970 3700 3906 3157 2087 3302 3297 2917 144 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 119(Y 1748 2787 1867 1117 3134 2580 896 2823 2430 2708 1928 1262 2616 482 1298 588 1603 2513 2720 2852 145 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 120(M 543 1502 454 655 1569 2005 725 1152 1280 1097 2617 860 2092 451 910 972 801 975 1856 1371 146 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 121(G 2913 3668 945 1911 5049 3566 2931 5265 3381 5155 4587 2191 3583 2748 4001 2853 3188 4499 4671 4461 147 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 122(E) 418 2234 1260 1822 2533 1769 443 2271 1177 485 1330 412 1871 9 542 693 747 1845 2428 811 148 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 123(V 2475 1721 4141 4120 2951 2548 3488 657 3886 2245 1927 3055 3187 3584 3760 1902 1766 2703 3796 3475 149 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 124(D 4232 4465 4158 2649 5412 3676 3575 6036 4125 5722 5419 2993 4216 3512 4617 4077 4401 5553 4696 4914 150 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 125(G 1595 2107 3720 3963 3970 3471 3570 3119 3981 3886 3293 2985 3155 3665 3837 1865 2027 946 4215 4068 151 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 126(M 2025 2478 4388 3797 1130 3739 2667 158 3453 1208 4105 3402 3608 2937 3236 2881 1961 880 2231 2028 152 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 127(W 150 757 49 299 277 651 839 485 392 910 599 300 401 623 162 99 38 448 1107 126 153 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 128(V 2447 1962 5103 4789 2553 4857 4827 1666 4719 1334 1279 4568 4656 4638 4849 4228 2447 3448 4233 3737 154 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 129(S) 2131 1670 3692 3910 4174 1935 3404 3963 3845 4238 3317 2599 2728 3415 3673 2964 1517 2788 4397 4269 155 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 130(G 4088 3924 4774 5139 5615 3825 4753 6303 5453 6014 5662 4812 4539 5232 5106 4370 4472 5527 4696 5561 156 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 131(A 3075 1760 2603 2695 3957 1941 2800 3736 2952 3942 3051 1370 2670 2640 3110 792 1502 2723 4139 3839 157 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 132(V 1970 1618 4439 3946 1865 3942 3147 1777 3693 528 807 3588 3866 3412 3626 3130 1942 2096 2880 2511 158 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 133(H 1102 2379 1738 1466 2663 2010 4358 2506 1383 2678 1892 969 2465 1337 1706 783 1293 2046 2891 2346 159 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 134(S) 419 1643 3250 3104 3814 1905 2857 3555 2981 3781 2885 2311 2298 2716 3080 2397 1614 2583 4014 3764 160 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 135(T) 1102 1714 3480 3700 4076 1973 3311 3843 3648 4140 3265 2565 2755 3310 3525 656 3729 2765 4307 4127 161 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 136(G 2443 1650 3680 3879 4195 2494 3389 3992 3844 4243 3304 2574 2707 3388 3682 672 1492 2786 4412 4299 162 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 137(D 2610 4414 3375 640 4704 2395 1885 4787 2296 4632 4069 2731 2988 1589 3253 2255 2733 4206 4710 3683 163 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 138(T) 1802 1610 4107 3628 1625 3434 2777 121 3310 979 612 3213 3529 3008 3231 2634 2771 1888 2588 2269 164 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 139(L) 2900 2445 5330 4886 1470 4921 4106 73 4643 2578 286 4689 4527 3948 4403 4253 2850 2224 3114 3047 165 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 140(R 4488 4181 4789 4318 5193 4148 3436 5568 2393 5152 4748 4156 4479 3286 4202 4614 4467 5273 4267 4649 166 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 141(P) 1695 1951 3539 3802 4288 2189 3492 4075 3870 4341 3515 2772 3709 3517 3739 1609 1819 3017 4404 4341 167 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 142(A 2997 1820 3672 3957 4327 2092 3531 4137 4026 4400 3498 2726 2859 3569 3834 1467 1684 2957 4469 4425 168 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 143(L) 3221 2748 5558 5092 1274 5134 4164 386 4812 3030 86 4956 4613 3913 4456 4503 3148 772 2987 3018 169 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 144(Q 2871 3512 2899 2161 4235 3278 1502 3869 191 3535 2901 2118 3368 4099 1519 2774 2657 3608 3317 3159 170 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 145(I) 2460 1967 5122 4804 2546 4899 4864 3420 4742 1315 1266 4595 4676 4657 4875 4271 2457 2106 4245 3757 171 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 146(I) 2482 1984 5127 4768 2335 4912 4643 3097 4684 313 1075 4568 4633 4487 4753 4252 2463 2302 3978 3593 172 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 147(K 2698 3306 2555 2385 4613 1689 2226 4379 3466 4176 3510 2406 3487 1898 1272 2718 2789 3870 3879 3804 173 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 148(T) 1775 2078 3737 3648 2269 2758 3026 1461 3198 577 1417 3004 3304 3108 3140 2144 3610 1504 3171 2809 174 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 149(K 1442 2243 1132 565 2592 2073 704 2218 2425 2266 1439 805 2179 1424 419 1063 1056 139 2490 1948 175 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 150(P) 2229 3531 751 745 4074 2482 1471 3788 860 3650 2934 1174 3599 1104 1174 2025 2220 3370 3701 3125 176 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 151(G 4088 3924 4774 5139 5615 3825 4753 6303 5453 6014 5662 4812 4539 5232 5106 4370 4472 5527 4696 5561 177 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 152(C 1031 2336 3000 2410 865 2361 1377 341 2075 407 99 2001 2486 1756 1986 688 649 1750 1361 1013 178 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 153(K 299 2345 751 248 2698 1882 2125 2424 2532 2376 1479 830 1996 103 500 327 894 1997 2541 1898 179 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 154(T) 908 953 2280 1684 935 2313 1108 722 344 1215 140 1590 2372 1223 1008 1350 1552 668 1392 1018 180 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 155(V 248 1426 4165 3634 1671 3597 2693 2073 3341 492 700 3234 3564 3048 3243 2750 488 2467 2499 2124 181 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 156(S) 1944 2458 3696 4011 4410 2644 3753 4675 4191 4807 4070 3198 3384 3886 4051 3656 2393 3615 4374 4307 182 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 157(G 693 1658 3600 3793 4209 2495 3362 4011 3794 4256 3314 2553 2706 3347 3658 2479 1495 2797 4419 4298 183 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 158(F) 757 848 3195 2588 2258 2533 1456 1163 2229 678 69 2151 2599 1889 2097 1006 949 1406 1371 1020 184 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 159(F) 3561 3044 5093 4992 4144 4702 2042 1417 4612 651 2173 3997 4516 3679 4183 4055 3507 1990 1263 230 185 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 160(I) 2399 1985 4894 4389 1240 4436 3536 2456 4148 1300 435 4090 4179 3651 3986 3652 2345 2073 2927 2716 186 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 161(M 2968 2569 5237 4809 1359 4763 3886 1091 4419 34 4848 4576 4450 3792 4170 4116 2930 673 2953 2839 187 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 162(Q 309 798 2699 2096 1118 2361 1203 789 1796 499 6 1823 2416 1904 1779 902 818 786 1249 892 188 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 163(V 884 1453 1331 759 1529 2054 754 426 1538 1352 623 938 708 503 609 1024 716 1556 1814 1348 189 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 164(P) 1205 2096 1371 1294 3530 1986 1732 3286 1522 3357 2495 1382 2600 1480 1930 689 1438 2587 3557 3013 190 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 165(D 1308 2864 2377 1708 3151 1969 847 2932 908 2865 1984 1026 2212 422 1139 1136 1274 2473 3039 1438 191 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 166(C 544 3117 563 1527 2011 1598 229 1884 984 1975 920 317 1617 1132 431 599 614 1564 1347 1372 192 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 167(E) 1135 2652 805 2212 2960 449 701 2724 352 2658 1760 534 2098 1133 1725 983 1089 2270 2825 2111 193 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 168(Y 3596 3426 4011 4236 756 3926 2017 3923 4119 3489 3396 3558 1274 3712 3935 3640 3783 3834 1423 4617 194 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 809 7942 1234 894 1115 701 1378 * * 169(G 2666 2751 3377 3700 4393 3753 3528 4841 4031 4741 4286 3394 3411 3824 3848 2912 3053 4080 3719 4290 195 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7148 8190 894 1115 2275 334 * * 170(H 909 2472 1648 1963 2749 1496 2279 2536 207 2473 1609 112 1774 16 789 911 884 2079 2655 1892 196 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7148 8190 894 1115 2275 334 * * 171(D 1659 3544 3062 1534 3720 1620 946 3633 1158 3508 2820 1807 2140 608 1992 1327 1718 3114 3702 2701 197 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7148 8190 894 1115 2275 334 * * 172(G 2666 2751 3377 3700 4393 3753 3528 4841 4031 4741 4286 3394 3411 3824 3848 2912 3053 4080 3719 4290 198 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 15 7148 8190 894 1115 1358 713 * * 173(M 896 558 2670 2060 1251 2185 1048 43 1731 665 2262 1717 2232 1411 1654 1259 1662 83 1007 665 199 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7414 8456 894 1115 1982 421 * * 174(L) 3012 2483 4991 4588 2563 4681 2567 365 4286 2641 286 4144 4140 3283 3862 3990 2889 1078 1633 952 200 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7414 8456 894 1115 249 2660 * * 175(V 1666 1535 3931 3496 1547 517 2575 46 3176 1766 593 3023 3348 2859 3069 2394 1705 2567 2418 2089 201 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 176(F) 4576 3559 5015 5343 4263 4881 1152 3449 4910 2776 2865 3550 4757 3679 4300 4154 4444 3622 401 2124 202 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 177(A 3345 1674 3727 3977 4175 1941 3439 3955 3910 4243 3329 2621 2737 3468 3709 825 1527 2788 4405 4282 203 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 178(W 150 757 49 299 277 651 839 485 392 910 599 300 401 623 162 99 38 448 1107 126 204 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 179(D 4232 4465 4158 2649 5412 3676 3575 6036 4125 5722 5419 2993 4216 3512 4617 4077 4401 5553 4696 4914 205 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 180(C 1236 5496 4197 4463 4068 2103 3603 3911 4139 4251 3414 2867 2893 3739 3846 647 1702 2864 4296 4138 206 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 181(A 2681 1648 3686 3867 4184 2067 3375 3979 3817 4229 3291 2570 2704 3369 3665 954 1488 2779 4401 4284 207 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 182(V 2422 1967 4993 4556 1946 4647 4007 2482 4386 447 1863 4301 4388 4024 4325 3914 2387 2602 3399 3128 208 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 183(N 1642 1630 3011 2702 1675 3027 2261 141 2480 399 827 3044 3220 2357 2577 2236 1678 2152 2448 1987 209 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 184(I) 1310 1421 2217 1185 1569 2673 1609 2204 1689 1167 660 1843 2154 1552 1934 1762 1280 885 2135 1720 210 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 185(N 257 1558 440 612 2481 1765 435 2211 31 2194 1330 2360 1863 1379 534 683 729 1796 2393 1727 211 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 186(P) 2875 3093 3978 4134 3333 3398 3665 3255 3879 167 2945 3727 4087 3894 3795 3160 3199 3243 3784 3483 212 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 187(T) 1617 2991 2432 518 3614 2086 1314 3413 1230 3383 2559 996 2497 937 1848 429 2498 2889 3583 2811 213 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 188(A 2241 1737 1506 513 2489 1943 1317 2071 1119 2312 1526 1230 2307 1010 1520 1714 1101 416 2692 2195 214 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 189(E) 525 2429 1776 1822 2741 1821 551 2497 176 2443 1530 453 124 1127 700 809 481 2048 2615 1917 215 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 190(Q 1567 3175 1704 1474 3449 2060 1054 3249 877 3174 2325 671 2377 2979 1481 1362 1084 2781 3353 2563 216 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 191(L) 3337 2808 5707 5127 1152 5377 4133 460 4878 2928 1801 5079 4636 3831 4452 4675 3207 697 2886 3022 217 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 192(A 3213 1669 3729 3963 4171 1937 3427 3953 3889 4236 3319 2614 2732 3450 3697 1430 1520 2784 4399 4276 218 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 193(E) 2200 4049 2514 2580 4247 2234 1519 4143 1674 4022 3301 1158 2729 1171 2473 318 2248 3627 4213 3243 219 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 194(I) 2474 2002 5104 4792 2432 4811 4734 3681 4697 1199 1182 4556 4627 4560 4788 4186 2477 1241 4098 3650 220 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 195(A 3609 2508 4184 4493 4592 2737 3989 4421 4496 4701 4051 3440 3474 4171 4253 2294 2487 3536 4484 4643 221 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 196(I) 1091 1076 2583 413 1145 2551 1453 2635 1815 911 317 1908 2625 1603 1923 64 352 1441 1667 1289 222 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 197(Q 518 2071 706 1236 2329 1787 464 158 79 2066 1190 460 1883 1523 571 1071 1153 1661 2309 1673 223 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 198(S) 690 1632 3665 3707 4013 1911 3224 3755 3549 4023 3114 2519 2683 3185 3470 2818 2263 2676 4240 4079 224 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 199(A 2957 1575 3607 3525 3508 456 2992 3142 3331 3468 2642 2460 2661 2989 3286 646 1410 731 3811 3585 225 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 200(E) 56 2745 1143 2154 3045 1930 765 2818 1875 2752 1859 945 2144 332 1009 1045 1166 2359 2923 2190 226 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 201(T) 1256 2015 1982 1726 3420 2098 1785 3082 927 3191 2379 1663 2604 1463 1433 524 3369 2475 3390 2984 227 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 202(W 2769 2207 4068 3807 2256 3458 1352 1888 3438 1847 1513 2925 3632 2881 3227 2665 2391 1867 3112 296 228 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 203(R 2470 3357 2623 1389 4114 3061 1112 3509 2409 3196 2485 1713 3049 686 2844 2333 2214 3234 3079 2887 229 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 540 7942 1697 894 1115 701 1378 * * 204(M 816 1388 782 226 1511 1668 358 1120 1679 119 1761 1028 1755 26 458 619 511 896 1709 1189 230 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7414 8456 894 1115 249 2660 * * 205(L) 2837 2376 5287 4745 1320 4877 3831 1365 4505 2429 1943 4548 4406 3738 4219 4120 2750 1537 2900 2889 231 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 206(C 1191 2897 3560 2991 2519 2720 1807 69 2623 837 305 2465 2840 2267 2460 1855 899 2022 1675 1297 232 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 207(G 1539 3140 2028 312 3421 2111 1027 3216 685 3138 2285 998 2361 1202 1409 1339 1520 2749 3312 2532 233 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 208(I) 334 1130 401 620 1157 2255 1040 2289 1141 979 1812 1358 2323 992 1401 1264 866 644 1591 1181 234 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 209(E) 425 2447 1685 2151 2749 1843 586 2497 831 2456 1553 477 1993 142 743 847 928 108 2639 1943 235 149 500 235 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 169 3204 8984 206 2913 701 1378 * * 210(P) 4497 4117 4899 5251 5560 4139 4798 6328 5454 5976 5741 5026 4302 5328 5104 4798 4844 5739 4665 5484 237 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 211(R 2896 3530 3730 2192 4456 3379 1195 3707 2481 3319 2662 2040 3291 1300 3145 2762 2535 3495 3134 3075 238 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 212(V 2838 2616 4834 4880 3213 3955 4416 610 4757 2310 2253 4383 4345 4643 4642 3732 3007 3763 4157 3886 239 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 213(A 3345 1674 3727 3977 4175 1941 3439 3955 3910 4243 3329 2621 2737 3468 3709 825 1527 2788 4405 4282 240 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 214(M 3386 2926 5503 5047 1187 5101 3953 644 4586 916 4893 4906 4599 3801 4244 4517 3301 1338 2841 2779 241 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 215(L) 4119 3542 5389 5357 2027 4767 4359 1609 5118 3293 979 5230 4771 4474 4724 5069 4106 2331 3412 3400 242 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 216(S) 1944 2458 3696 4011 4410 2644 3753 4675 4191 4807 4070 3198 3384 3886 4051 3656 2393 3615 4374 4307 243 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 217(F) 4596 3564 5022 5356 4100 4892 1140 3474 4920 2799 2888 3546 4763 3678 4303 4158 4459 3642 389 2808 244 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 218(S) 1944 2458 3696 4011 4410 2644 3753 4675 4191 4807 4070 3198 3384 3886 4051 3656 2393 3615 4374 4307 245 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 219(T) 1086 1700 3489 3680 4071 1960 3290 3842 3622 4127 3243 2550 2741 3278 3511 1633 3501 2755 4299 4122 246 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 220(K 1211 2188 1252 684 2201 2183 735 2041 2458 2110 2342 1770 2264 372 433 1182 1128 1772 2263 1110 247 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 221(G 4088 3924 4774 5139 5615 3825 4753 6303 5453 6014 5662 4812 4539 5232 5106 4370 4472 5527 4696 5561 248 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 222(S) 1944 2458 3696 4011 4410 2644 3753 4675 4191 4807 4070 3198 3384 3886 4051 3656 2393 3615 4374 4307 249 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 223(A 3609 2508 4184 4493 4592 2737 3989 4421 4496 4701 4051 3440 3474 4171 4253 2294 2487 3536 4484 4643 250 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 224(K 2345 3215 2680 1702 4004 2994 1114 3430 3145 3150 2429 1707 3005 690 1537 1210 2130 3134 3052 2847 251 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 225(H 1041 1842 1795 1535 3225 1554 2940 2958 1555 3088 2233 1512 2445 1442 1925 2143 928 2312 3333 2843 252 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 226(P) 1812 3493 1638 2272 3746 2139 1234 3572 714 3480 2668 741 2320 848 1793 1570 1815 3091 3658 2819 253 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 227(M 1063 1672 1185 1731 1647 2144 910 1218 725 1142 2726 1099 2261 651 1101 1179 1008 1100 2011 1532 254 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 228(W 150 757 49 299 277 651 839 485 392 910 599 300 401 623 162 99 38 448 1107 126 255 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 229(V 1683 1779 4225 4114 2704 2952 3573 161 3855 1868 1633 3296 3474 3653 3791 2302 2073 3159 3712 3334 256 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 230(E) 576 3124 2215 2294 3448 2061 1092 3240 936 3183 2340 696 2393 692 1547 1375 1093 2772 3374 2589 257 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 231(K 315 2794 1360 1079 3606 2397 1201 3242 3269 3135 2344 1290 2657 798 553 1655 1723 2800 3186 2733 258 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 232(V 1806 1830 4356 4254 2729 3152 3789 6 4016 1803 1616 3476 3637 3839 3966 2513 660 3495 3859 3459 259 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 233(Q 793 1527 1108 552 1618 1947 1457 1157 637 1440 683 772 2030 1773 839 896 518 491 1868 1365 260 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 234(E) 300 2398 695 1659 2713 1806 518 2470 1395 2411 1494 1531 1936 1405 648 775 852 2018 2579 1883 261 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 235(A 3370 1675 3725 3979 4176 1943 3442 3956 3915 4245 3332 2623 2739 3473 3712 667 1529 2789 4406 4283 262 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 236(T) 1487 1747 4009 3951 2829 2596 3358 658 3663 2137 1837 3024 3208 3442 3576 1949 3247 2128 3661 3318 263 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 237(K 78 2444 636 1476 2774 1872 554 2516 2347 2450 1542 1061 1996 1188 567 849 918 2073 2605 1935 264 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 238(L) 2340 2043 4429 3825 1249 4051 2880 2209 47 2445 154 3576 3849 2947 3012 3227 2264 489 2490 2321 265 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 239(A 3066 1532 3416 3204 2546 2126 2613 1745 2919 251 1678 2434 2743 2680 2903 1429 721 1481 3078 2775 266 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 240(K 2775 3478 3431 2065 4356 3299 1170 3648 3068 3281 2609 1958 3227 2481 1606 2643 2444 3419 3113 3021 267 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 241(E) 5 3248 527 3274 3691 2238 1242 3429 736 3351 2559 905 2586 855 1282 1630 1816 2982 3491 2801 268 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 242(R 1203 1274 1535 960 714 2112 836 874 1242 45 1502 1087 2184 670 1607 1097 814 732 1655 1220 269 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 243(A 1867 2550 755 648 2859 1875 646 2618 291 2563 1661 1632 2046 205 1386 915 1009 2170 2737 2031 270 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 244(P) 1833 2267 3769 3944 3591 2645 3527 2530 3805 3285 2936 3132 3954 3636 3703 2130 2228 835 4002 3761 271 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 245(D 206 2972 2710 1351 3255 2000 1755 3042 683 2972 2100 614 2269 1462 1262 1213 1369 2579 3148 2386 272 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 246(L) 3414 2874 5769 5161 1383 5422 4072 994 4912 2728 2007 5118 4630 3791 4438 4708 3264 1344 2810 2947 273 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 247(A 1831 2029 575 204 2274 1806 494 1962 927 82 1160 494 270 72 607 735 508 1613 2285 1663 274 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 248(I) 2431 2000 4947 4466 1231 4532 3688 3189 4249 517 529 4183 4266 3781 4114 3766 2383 1363 3060 2816 275 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 249(D 4232 4465 4158 2649 5412 3676 3575 6036 4125 5722 5419 2993 4216 3512 4617 4077 4401 5553 4696 4914 276 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 250(G 4088 3924 4774 5139 5615 3825 4753 6303 5453 6014 5662 4812 4539 5232 5106 4370 4472 5527 4696 5561 277 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 251(E) 4205 4417 2343 3901 5349 3698 3534 5837 3838 5561 5237 3039 4218 3453 4201 4072 4352 5409 4640 4860 278 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 252(L) 3602 3015 5919 5328 1158 5654 4138 616 5093 2938 2013 5330 4744 3877 4575 4984 3438 1455 2794 2838 279 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 253(Q 4207 4117 3531 3731 4752 3930 3721 5554 3395 5179 4897 3823 4399 4556 3405 4301 4380 5208 4301 4384 280 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 254(F) 2863 2433 5172 4640 3169 4702 3234 392 4348 1785 84 4307 4290 3544 4021 3916 2769 1504 2393 2005 281 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 255(D 4232 4465 4158 2649 5412 3676 3575 6036 4125 5722 5419 2993 4216 3512 4617 4077 4401 5553 4696 4914 282 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 256(A 3609 2508 4184 4493 4592 2737 3989 4421 4496 4701 4051 3440 3474 4171 4253 2294 2487 3536 4484 4643 283 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 257(A 3186 1669 3728 3961 4171 1936 3425 3953 3886 4235 3318 2613 2731 3447 3695 1524 1520 2783 4398 4275 284 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 258(F) 3002 2525 5316 4818 3177 4924 3350 2613 4551 818 147 4496 4441 3703 4215 4169 2905 737 2444 1989 285 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 259(V 2294 2438 724 2485 3071 3156 3064 720 3372 2390 2164 2598 3629 2975 3716 2757 2460 3538 3861 3370 286 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 260(P) 1594 2934 619 588 3418 2163 1118 3148 755 3092 2272 870 3320 720 1108 321 1590 2706 3251 2595 287 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 261(E) 232 2432 633 1887 2799 1880 622 2541 1768 2491 1591 543 2032 178 679 1460 962 2096 2660 1989 288 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 262(V 2838 2616 4834 4880 3213 3955 4416 610 4757 2310 2253 4383 4345 4643 4642 3732 3007 3763 4157 3886 289 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 263(A 3303 1827 3692 3978 4328 1129 3541 4137 4037 4403 3503 2739 2867 3583 3840 1477 1693 2962 4468 4428 290 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 264(E) 1471 2341 669 1713 2646 1806 509 2389 1384 680 1440 444 1930 60 624 766 833 1955 2531 1848 291 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 265(Q 236 2063 853 302 2314 1882 542 1988 1473 779 1199 578 1977 1845 534 1012 820 1652 2307 1712 292 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 266(K 4125 4091 4017 3615 5101 3960 2958 5182 3972 4825 4345 3552 4249 2709 1809 4154 4044 4895 4132 4389 293 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 267(A 3348 2109 3336 3412 1764 2509 2437 2956 3286 3118 2630 2708 3150 3050 3249 1903 2016 2533 2364 1487 294 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 268(P) 1528 2315 1720 1687 3833 2233 1895 3559 797 3582 2793 1690 3653 1574 1445 406 1768 2881 3673 3270 295 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 269(G 1935 3089 2442 885 4418 2762 1974 4330 2285 4310 3586 1182 2821 1681 3048 394 2183 3549 4481 3646 296 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 270(S) 1944 2458 3696 4011 4410 2644 3753 4675 4191 4807 4070 3198 3384 3886 4051 3656 2393 3615 4374 4307 297 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 271(K 986 2487 579 781 2798 1840 584 2559 1709 2499 1588 1699 1485 137 743 394 935 2105 2668 1964 298 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 272(V 2488 1997 5115 4741 2236 4876 4521 1399 4638 586 992 4534 4596 4394 4675 4204 2466 3282 3849 3500 299 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 273(A 2860 2613 1445 1105 3345 2365 1181 2968 911 2930 2150 1269 2615 1502 562 1587 1627 2570 3048 2602 300 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 274(G 4088 3924 4774 5139 5615 3825 4753 6303 5453 6014 5662 4812 4539 5232 5106 4370 4472 5527 4696 5561 301 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 275(H 1098 2519 644 333 2880 1987 2767 2597 2281 2509 1617 857 2093 153 1114 970 1024 2167 2636 2015 302 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 276(A 3609 2508 4184 4493 4592 2737 3989 4421 4496 4701 4051 3440 3474 4171 4253 2294 2487 3536 4484 4643 303 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 277(N 1586 2305 1738 2020 4028 2232 2587 3973 2635 4158 3406 4054 2920 2402 2900 1720 681 3116 4157 3676 304 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 278(V 2446 1963 5100 4786 2548 4847 4816 1583 4714 1330 1276 4562 4651 4631 4841 4218 2447 3468 4224 3729 305 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 279(W 150 757 49 299 277 651 839 485 392 910 599 300 401 623 162 99 38 448 1107 126 306 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 280(F) 3486 2932 5765 5195 3114 5447 3820 632 4937 2173 2384 5073 4656 3798 4452 4740 3334 1436 2628 2474 307 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 281(I) 2458 1961 5125 4807 2569 4914 4890 3219 4751 1339 1284 4603 4685 4676 4892 4286 2453 2471 4276 3778 308 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 282(F) 4720 3927 5139 5454 4513 4504 2520 3938 5326 3307 3429 4469 4782 4575 4818 4787 4810 4172 1812 736 309 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 283(P) 4497 4117 4899 5251 5560 4139 4798 6328 5454 5976 5741 5026 4302 5328 5104 4798 4844 5739 4665 5484 310 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 284(D 2322 4185 3035 1469 4418 2266 1625 4341 1869 4214 3539 867 2800 1293 2724 1786 2396 3805 4409 3395 311 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 285(L) 3492 2925 5831 5302 1190 5571 4334 974 5050 3059 17 5312 4763 3965 4606 4957 3364 1095 2972 3089 312 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 286(Q 2004 3769 1173 2079 3991 2187 1375 3849 1403 3743 2969 1136 2626 3177 2117 1727 2027 3352 3926 3026 313 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 287(A 2861 2401 3941 3933 3713 1915 3209 3405 3647 3703 2846 2573 2679 3249 3498 1246 795 2497 4004 3830 314 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 288(G 804 1917 3555 3872 4406 3488 3564 4245 4053 4497 3611 2768 2934 3603 3880 1567 1787 3064 4502 4481 315 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 289(N 3642 3838 3004 3356 4743 3612 3773 5643 4099 5460 5060 4378 4185 3865 4225 3744 3950 5002 4391 4381 316 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 290(I) 3202 2787 5004 4996 2836 4384 4440 3937 4838 1815 1871 4704 4600 4683 4704 4353 3280 603 3938 3661 317 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 291(G 1452 1874 3596 3903 4376 3335 3550 4203 4044 4459 3565 2747 2900 3588 3862 1522 1740 3018 4492 4461 318 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 292(Y 4791 3935 4914 5231 815 4530 2178 4357 5024 3745 3807 4206 4772 4328 4584 4677 4837 4435 1477 4857 319 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 293(K 4125 4091 4017 3615 5101 3960 2958 5182 3972 4825 4345 3552 4249 2709 1809 4154 4044 4895 4132 4389 320 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 294(I) 3130 2603 5572 5096 1392 5301 4317 3061 4881 2084 185 5010 4688 4034 4578 4656 3046 463 3127 3143 321 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 295(A 2675 1535 3836 3683 2774 2214 2959 1215 3410 2393 1857 2670 2859 3085 3310 1533 792 2057 3350 3057 322 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 296(Q 4207 4117 3531 3731 4752 3930 3721 5554 3395 5179 4897 3823 4399 4556 3405 4301 4380 5208 4301 4384 323 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 297(R 4488 4181 4789 4318 5193 4148 3436 5568 2393 5152 4748 4156 4479 3286 4202 4614 4467 5273 4267 4649 324 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 298(F) 3500 2941 5773 5206 3166 5461 3812 635 4949 2227 2003 5083 4665 3805 4462 4757 3346 1441 2619 2449 325 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 299(G 2279 1650 3675 3870 4196 2606 3385 3993 3836 4243 3303 2572 2706 3382 3678 828 1491 2786 4412 4298 326 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 300(K 1571 2888 927 655 3321 2063 1468 3019 2355 2910 2082 920 2447 523 551 1431 1504 2612 2998 2431 327 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 301(F) 2448 2615 4303 4172 2956 3922 1214 2324 3828 2183 1926 3107 3996 3105 3543 3119 2926 2325 560 2239 328 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 302(E) 1402 2920 1061 2832 3242 2054 879 2998 774 2910 2042 672 2284 454 1073 1230 1357 2549 3055 2355 329 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 303(A 3609 2508 4184 4493 4592 2737 3989 4421 4496 4701 4051 3440 3474 4171 4253 2294 2487 3536 4484 4643 330 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 304(I) 1878 1552 4312 3775 1491 3734 2739 2447 3471 521 541 3367 3656 3110 3335 2889 1832 2126 2430 2171 331 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 305(G 4088 3924 4774 5139 5615 3825 4753 6303 5453 6014 5662 4812 4539 5232 5106 4370 4472 5527 4696 5561 332 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 306(P) 4497 4117 4899 5251 5560 4139 4798 6328 5454 5976 5741 5026 4302 5328 5104 4798 4844 5739 4665 5484 333 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 307(I) 2574 2082 5179 4778 2009 4923 4424 3104 4656 1327 776 4584 4587 4278 4620 4242 2542 1595 3640 3399 334 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 308(C 1490 2918 3842 3258 1170 3142 2133 1934 2902 1595 258 2806 3152 2528 2741 2266 1394 65 1929 1611 335 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 309(Q 4207 4117 3531 3731 4752 3930 3721 5554 3395 5179 4897 3823 4399 4556 3405 4301 4380 5208 4301 4384 336 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 310(G 4088 3924 4774 5139 5615 3825 4753 6303 5453 6014 5662 4812 4539 5232 5106 4370 4472 5527 4696 5561 337 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 311(L) 3686 3084 5727 5319 2036 5453 3331 789 5052 2929 88 4966 4743 3883 4542 4837 3542 1589 2266 1680 338 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 312(A 2898 1987 2240 2485 4049 2085 2826 3885 3004 4117 3288 2707 2813 2683 3186 1506 1726 2917 4217 3861 339 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 313(K 1066 2626 1795 1052 3050 2498 872 2618 2866 2563 1526 1206 2543 458 1142 1556 1494 2321 2682 2281 340 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 314(P) 4497 4117 4899 5251 5560 4139 4798 6328 5454 5976 5741 5026 4302 5328 5104 4798 4844 5739 4665 5484 341 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 315(V 2475 1980 5117 4752 2317 4893 4598 2349 4663 557 1062 4548 4617 4458 4725 4228 2455 2932 3943 3564 342 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 316(H 173 2262 944 767 3091 1973 2974 2865 947 2905 2052 2949 2338 840 1414 1020 1292 2346 3104 2474 343 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 317(D 4232 4465 4158 2649 5412 3676 3575 6036 4125 5722 5419 2993 4216 3512 4617 4077 4401 5553 4696 4914 344 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 318(L) 4119 3542 5389 5357 2027 4767 4359 1609 5118 3293 979 5230 4771 4474 4724 5069 4106 2331 3412 3400 345 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 319(S) 1944 2458 3696 4011 4410 2644 3753 4675 4191 4807 4070 3198 3384 3886 4051 3656 2393 3615 4374 4307 346 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 320(R 4488 4181 4789 4318 5193 4148 3436 5568 2393 5152 4748 4156 4479 3286 4202 4614 4467 5273 4267 4649 347 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 321(G 4088 3924 4774 5139 5615 3825 4753 6303 5453 6014 5662 4812 4539 5232 5106 4370 4472 5527 4696 5561 348 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 322(C 1383 5545 4327 4558 4019 2243 3671 3414 4157 3975 3301 3006 3016 3818 3871 1656 540 2675 4269 4170 349 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 323(S) 1998 3389 920 646 4194 2202 1688 4095 1857 4037 3301 1873 2745 1360 2596 2972 2145 3473 4215 3333 350 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 324(A 1796 2218 759 1121 2509 1874 667 2198 343 2259 1405 578 2039 254 849 333 937 1127 2521 1880 351 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 325(E) 2264 4130 1161 3061 4316 2257 1556 4221 1720 4092 3390 846 2763 2340 2512 1940 2316 3705 4277 3299 352 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 326(D 33 4335 3607 1205 4570 2294 1708 4511 2035 4382 3756 897 2858 1390 2952 2084 2528 3963 4583 3525 353 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 327(I) 2456 1959 5125 4808 2580 4919 4900 3012 4754 1350 1292 4605 4688 4684 4898 4291 2452 2731 4288 3786 354 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 328(Y 2065 1703 4458 4000 1497 3978 2824 1512 3720 1096 877 3567 3903 3404 3627 3169 2033 2744 2424 2949 355 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 329(K 1420 3005 1129 1097 3290 1087 935 3080 2093 3005 2135 1724 2283 517 1280 1233 1393 2613 3178 2412 356 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 330(W 150 757 49 299 277 651 839 485 392 910 599 300 401 623 162 99 38 448 1107 126 357 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 331(V 242 1739 4505 4069 1901 3865 3328 339 3811 1145 812 3662 3892 3515 3741 3100 2045 3053 3031 2693 358 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 332(A 2785 1548 3757 3706 3183 2010 3016 2274 3464 3018 2323 2559 2725 3105 3346 519 1436 1956 3604 3353 359 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 333(A 2301 1785 4329 3814 1389 3715 2881 1608 3501 1601 328 3440 3685 3079 3364 2901 2013 142 2516 2281 360 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 334(I) 2461 1969 5121 4803 2539 4894 4855 3457 4740 1307 1260 4593 4673 4650 4870 4267 2458 2020 4235 3750 361 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 335(T) 1694 1679 3770 3955 4023 1971 3381 3579 3783 3994 3161 2633 2754 3407 3608 1339 3474 2625 4286 4150 362 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 336(A 3145 2363 4124 4092 3068 2102 3176 1748 3745 2761 2179 2727 2812 3341 3530 1439 1489 622 3594 3345 363 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 337(V 1549 1561 3980 3800 2617 2450 3066 638 3529 2084 1647 2851 3038 3217 3427 1764 645 3033 3323 3003 364 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 338(Q 1998 3480 470 1001 3872 2292 1373 3668 1085 3566 2804 947 1060 3783 1533 1779 2007 3214 3690 2964 365 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 339(A 2845 1741 2441 2116 3476 181 2133 3194 1899 3346 2475 1852 2527 1850 1122 752 1333 2425 3579 3191 366 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 9 7942 8984 894 1115 701 1378 * * 340(Q 288 1371 2820 2256 1089 2819 1668 1024 1933 1596 165 2151 2848 2772 2022 1891 1329 469 1817 1496 367 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 444 7942 1940 894 1115 701 1378 * * 341(A 2553 1499 1618 1106 1437 1994 884 1437 516 1637 947 1150 2220 717 1070 1116 989 1219 1772 1598 368 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 154 7510 3381 894 1115 1844 471 * * 342(Q 840 2183 2051 46 2487 478 471 2205 189 2230 1379 273 1821 2234 719 721 813 565 2466 1782 369 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 13 7368 8411 894 1115 2041 402 * * 343(K 542 1518 903 68 1686 266 294 542 1621 1463 666 332 1689 78 434 539 484 817 1827 1263 370 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 435 7368 1975 894 1115 2041 402 * * 344(K 842 1833 1221 299 2188 1329 107 1924 1349 1907 1015 8 1492 329 247 1153 413 1495 2105 1431 371 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 1110 6951 919 894 1115 2432 296 * * 345(R 956 1423 1186 681 1830 1454 276 1686 690 1640 1095 694 1696 14 3253 1003 929 1464 1574 1319 372 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 37 5878 6920 894 1115 2888 209 * * 346(E) 804 1735 497 2874 2096 1058 327 1858 157 1967 1380 40 1471 39 568 688 853 1562 2027 1547 373 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 37 5878 6920 894 1115 2888 209 * * 347(R 956 1423 1186 681 1830 1454 276 1686 690 1640 1095 694 1696 14 3253 1003 929 1464 1574 1319 374 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 37 5878 6920 894 1115 2888 209 * * 348(S) 227 390 660 668 1564 565 762 1409 705 1690 1020 460 1182 625 900 2604 94 861 1818 1338 375 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 37 5878 6920 894 1115 2888 209 * * 349(L) 1162 1035 2273 2015 35 2243 1374 599 1587 2466 801 1903 2350 1486 1587 1745 1181 283 1141 718 376 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 37 5878 6920 894 1115 2888 209 * * 350(H 948 1353 590 533 353 1374 4354 1609 210 1544 1090 628 1716 415 369 985 1004 1409 758 74 377 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 37 5878 6920 894 1115 2888 209 * * 351(T) 71 435 987 905 1386 761 851 654 738 1152 667 673 1327 733 889 159 2865 370 1730 1351 378 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 37 5878 6920 894 1115 2888 209 * * 352(L) 1162 1035 2273 2015 35 2243 1374 599 1587 2466 801 1903 2350 1486 1587 1745 1181 283 1141 718 379 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 37 5878 6920 894 1115 2888 209 * * 353(V 558 557 2112 1895 730 1799 1509 1150 1619 15 149 1639 2121 1558 1693 1183 681 2778 1647 1187 380 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 37 5878 6920 894 1115 2888 209 * * 354(E) 804 1735 497 2874 2096 1058 327 1858 157 1967 1380 40 1471 39 568 688 853 1562 2027 1547 381 149 500 233 43 381 399 106 626 210 466 720 275 394 45 96 359 117 369 294 249 37 5878 6920 894 1115 127 3565 * * 355(W 150 757 49 299 277 651 839 485 392 910 599 300 401 623 162 99 38 448 1107 126 382 * * * * * * * * * * * * * * * * * * * * * * * * * * * * 0