Biological Production of Multi-Carbon Compounds from Methane
20240368638 ยท 2024-11-07
Inventors
- William J. Coleman (Redwood City, CA, US)
- Genevieve M. Vidanes (San Francisco, CA, US)
- Guillaume Cottarel (Mountain View, CA, US)
- Sheela MULEY (Fremont, CA, US)
- Roy Kamimura (Daly City, CA, US)
- Akbar F. Javan (Chapel Hill, NC, US)
- Jianping Sun (Belmont, CA, US)
- Eli S. GROBAN (San Francisco, CA, US)
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
C12Y401/01072
CHEMISTRY; METALLURGY
C12N9/1022
CHEMISTRY; METALLURGY
C12Y402/01033
CHEMISTRY; METALLURGY
C12Y402/01009
CHEMISTRY; METALLURGY
International classification
Abstract
Multi-carbon compounds such as ethanol, n-butanol, sec-butanol, isobutanol, tert-butanol, fatty (or aliphatic long chain) alcohols, fatty acid methyl esters, 2,3-butanediol and the like, are important industrial commodity chemicals with a variety of applications. The present invention provides metabolically engineered host microorganisms which metabolize methane (CH.sub.4) as their sole carbon source to produce multi-carbon compounds for use in fuels (e.g., bio-fuel, bio-diesel) and bio-based chemicals. Furthermore, use of the metabolically engineered host microorganisms of the invention (which utilize methane as the sole carbon source) mitigate current industry practices and methods of producing multi-carbon compounds from petroleum or petroleum-derived feedstocks, and ameliorate much of the ongoing depletion of arable food source farmland currently being diverted to grow bio-fuel feedstocks, and as such, improve the environmental footprint of future bio-fuel, bio-diesel and bio-based chemical compositions.
Claims
1-90. (canceled)
91. A genetically modified methanotroph comprising a heterologous polynucleotide encoding for an acetolactate synthase (ALS), wherein the acetolactate synthase can catalyze the conversion of pyruvate to acetolactate and comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO: 2, and wherein said methanotroph is capable of converting formaldehyde to pyruvate through a type I RuMP pathway or a type II serine pathway.
92. The methanotroph of claim 91, wherein the methanotroph further comprises a heterologous polynucleotide encoding a ketoacid decarboxylase (KDC), wherein the ketoacid decarboxylase can catalyze the conversion of ketoisovalerate to isobutryaldehyde.
93. The methanotroph of claim 92, wherein the methanotroph further comprises a a heterologous polynucleotide encoding a ketol-acid reductoisomerase (KARI), a heterologous polynucleotide encoding a dihydroxy-acid dehydratase (DHAD), and a heterologous polynucleotide encoding an alcohol dehydrogenase (ADH); wherein the ketol-acid reductoisomerase can catalyze the conversion of acetolactate to 2,3-dihydroxyisovalerate, wherein the dihydroxy-acid dehydratase can catalyze the conversion of 2,3-dihydroxyisovalerate to ketoisovalerate, and wherein the alcohol dehydrogenase can catalyze the conversion of isobutyraldehyde to isobutanol.
94. The methanotroph of claim 93, wherein the methanotroph further comprises a heterologous polynucleotide encoding an alcohol dehydrogenase (ADH), wherein the ketoacid decarboxylase can catalyze the conversion of isobutyraldehyde to isobutanol.
95. The methanotroph of claim 94, wherein the ketoacid decarboxylase (KDC) comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO: 8.
96. The methanotroph of claim 91, wherein the methanotroph further comprises a a heterologous polynucleotide encoding a ketol-acid reductoisomerase (KARI), heterologous polynucleotide encoding a dihydroxy-acid dehydratase (DHAD), and a heterologous polynucleotide encoding a ketoacid decarboxylase (KDC); and a heterologous polynucleotide encoding an alcohol dehydrogenase (ADH); wherein the acetolactate synthase can catalyze the conversion of pyruvate to acetolactate and comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO: 2, wherein the ketol-acid reductoisomerase can catalyze the conversion of acetolactate to 2,3-dihydroxyisovalerate and comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO: 4, wherein the dihydroxy-acid dehydratase can catalyze the conversion of 2,3-dihydroxyisovalerate to ketoisovalerate and comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO: 6, wherein the ketoacid decarboxylase can catalyze the conversion of ketoisovalerate to isobutryaldehyde and comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO: 8, and wherein the alcohol dehydrogenase can catalyze the conversion of isobutyraldehyde to isobutanol and comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO: 10.
97. The methanotroph of claim 96, wherein the methanotroph further comprises a polynucleotide sequence encoding for the alcohol dehydrogenase (ADH) and a promoter, wherein said promoter can direct the expression of the alcohol dehydrogenase in the methanotroph.
98. The methanotroph of claim 92, wherein the methanotroph further comprises a polynucleotide sequence encoding for the ketoacid decarboxylase (KDC), the alcohol dehydrogenase (ADH), and a promoter, wherein said promoter can direct the expression of the ketoacid decarboxylase and the alcohol dehydrogenase (ADH) in the methanotroph.
99. The methanotroph of claim 93, wherein the methanotroph further comprises a polynucleotide sequence encoding for the acetolactate synthase (ALS), the ketol-acid reductoisomerase (KARI), the dihydroxy-acid dehydratase (DHAD), the ketoacid decarboxylase (KDC), the alcohol dehydrogenase (ADH) and a promoter, wherein said promoter can direct the expression of the acetolactate synthase (ALS), the ketol-acid reductoisomerase (KARI), the dihydroxy-acid dehydratase (DHAD), the ketoacid decarboxylase (KDC), and the alcohol dehydrogenase (ADH) in a methanotroph.
100. The methanotroph of claim 99, wherein said promoter is constitutive.
101. The methanotroph of claim 99, wherein said promoter is inducible.
102. The methanotroph of claim 91, wherein said methanotroph is from the genus Methylobacter, Methylomicrobium, Methylomonas, Methylocaldum, Methylococcus, Methylosoma, Methylosarcina, Methylothermus, Methylohalobius, Methylogaea, Methylovulum, Crenothrix, Clonothrix, Methylosphaera, Methylocapsa, Methylocella, Methylosinus, Methylocystis, or Methyloacidophilum.
103. The methanotroph of claim 91, wherein said methanotroph is from the genus Methylococcus.
104. The methanotroph of claim 91, wherein said methanotroph is from the species Methylococcus capsulatus.
105. The methanotroph of claim 91, wherein said methanotroph is from the strain Methylococcus capsulatus strain Bath.
106. A method of making a multi-carbon compound comprising: (a) contacting a genetically modified methanotroph with a multi-carbon product precursor comprising a heterologous polynucleotide encoding for an acetolactate synthase (ALS), wherein the ALS can catalyze the conversion of pyruvate to acetolactate and comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO: 2, and wherein said methanotroph is capable of converting formaldehyde to pyruvate through a type I RuMP pathway or a type II serine pathway; and (b) growing said methanotroph in conditions to produce a multi-carbon compound.
107. The method of claim 105 wherein said precursor is methane.
108. The method of claim 106 wherein said multi-carbon compound is isobutanol.
109. The method of claim 106 wherein said multi-carbon product is 1-butanol.
110. A genetically modified methanotroph capable of converting methane to a multi-carbon product comprising a heterologous polynucleotide encoding for an acetolactate synthase (ALS), wherein the ALS has acetolactate synthase can catalyze the conversion of pyruvate to acetolactate and comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO: 2, wherein said methanotroph is capable of converting formaldehyde to pyruvate through a type I RuMP pathway or a type II serine pathway.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
[0064] Certain embodiments of the invention are illustrated in the drawings, in which:
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DETAILED DESCRIPTION OF THE INVENTION
[0074] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which this disclosure pertains. Although methods and materials similar or equivalent to those described herein can be used in the practice of the disclosed methods and compositions, the exemplary methods, devices and materials are described herein.
[0075] In certain embodiments, the present invention is directed to host microorganisms metabolically engineered to produce multi-carbon compounds. Multi-carbon compounds such as ethanol, n-butanol, sec-butanol, isobutanol, tert-butanol, fatty (or aliphatic long chain) alcohols, fatty acid methyl esters, 2,3-butanediol and the like, are important industrial commodity chemicals with a variety of applications, including, but not limited to their use in fuels (e.g., bio-fuel, bio-diesel) and bio-based chemicals. The present invention addresses a number of commercial, industrial and environmental needs in the art related to the production of multi-carbon compounds.
[0076] As set forth herein, the metabolically engineered host microorganisms of the present invention utilize methane (CH.sub.4) as their sole carbon source (i.e., the host microorganism does not require plant based feedstocks for growth and energy) and ameliorate much of the ongoing depletion of arable food source farmland currently being diverted to grow bio-fuel feedstocks, and as such, improve the environmental footprint of future bio-fuel, bio-diesel and bio-based chemical compositions. Furthermore, use of the metabolically engineered host microorganisms set forth in the present invention (which utilize methane as the sole carbon source) mitigate current industry practices and methods of producing multi-carbon compounds from petroleum or petroleum-derived feedstocks.
[0077] Thus, in certain embodiments of the invention, a host microorganism is genetically engineered to produce multi-carbon compounds. As is known in the art, methanotrophic organisms are able to metabolize methane as their primary source of carbon and energy, can grow aerobically or anaerobically, and require single-carbon compounds (e.g., methane, CH.sub.4; methanol, CH.sub.3OH and/or formaldehyde, H.sub.2CO) to survive. In particular embodiments, a host microorganism of the invention is a methanotroph. As defined herein, a methanotroph, a methanotrophic or a methanophile host microorganism of the invention is a prokaryotic microorganism which can metabolize methane as its primary source of carbon and energy.
[0078] In other embodiments, the host microorganism of the invention is a non-methanotrophic microorganism genetically engineered to metabolize methane as its only source of carbon and energy. As defined herein, a non-methanotroph host microorganism of the invention is a host microorganism which cannot metabolize (or utilize) methane as its sole carbon source, until the non-methanotroph host microorganism has been genetically modified or engineered according to the methods of the present invention. As further defined herein, a non-methanotroph host microorganism of the invention includes any prokaryotic and eukaryotic microbial species which comprise a complete or partial endogenous ribulose monophosphate (RuMP) pathway, a serine pathway or a mixed RuMP/serine pathway (e.g., see RuMP, serine and mixed (Type X) pathways described below). In certain embodiments, a non-methanotroph host microorganism of the invention includes prokaryotic and eukaryotic microbial species from the Domains Archaea, Bacteria and Eucarya, wherein the Domain Eucarya includes yeast, filamentous fungi, protozoa, algae or higher Protista. The terms microbial and microbes are used interchangeably with the term microorganism.
[0079] As defined herein, the phrase providing a methanotrophic host microorganism that metabolizes methane to methanol and metabolizes methanol to formaldehyde refers to an endogenous enzymatic activity encoded by one or more endogenous genes of the methanotroph host microorganism. For example, an endogenous enzyme (or polypeptide) encoded by one or more endogenous genes of a methanotroph host microorganism include a methane monooxygenase (MMO) enzyme (which metabolizes (or converts) methane to methanol) and a methanol dehydrogenase (MDH) enzyme (which metabolizes (or converts) methanol to formaldehyde). Stated another way, the phrase providing a methanotrophic host microorganism that metabolizes methane to methanol and metabolizes methanol to formaldehyde does not require the introduction of exogenous (or heterologous) genes encoding single-carbon (C1) oxidizing enzymes (or polypeptides), as such enzymes and the activity thereof are inherent (endogenous) attributes of a methanotrophic host microorganism of the invention.
[0080] Furthermore, as is known in the art, a methanotrophic host microorganism of the invention comprises endogenous genes encoding at least a Type I methanotroph RuMP pathway and/or a Type II methanotroph serine pathway. In general, Type I methanotrophs (e.g., Methylomonas, Methylomicrobium, Methylobacter, Methylocaldum, Methylosphaera) assimilate formaldehyde produced (i.e., from the oxidation of methane to methanol and methanol to formaldehyde), using the ribulose monophosphate pathway (RuMP), whereas Type II methanotrophs (e.g., Methylocystis and Methylosinus) assimilate formaldehyde produced (i.e., from the oxidation of methane to methanol and methanol to formaldehyde), using the serine pathway. Lastly, the genus Methylococcus are known to comprise a combination of characteristics of both Type I methanotroph (RuMP) pathway and Type II methanotroph (serine) pathway.
[0081] The ribulose monophosphate pathway (RuMP) was originally identified in methanotrophic bacteria, as described above. However, more recent genome sequence analysis of various microorganisms have revealed that the key enzymes of the RuMP pathway (e.g., 3-hexulose-6-phosphate (HPS), 6-phsopho-3-hexuloisomerase (PHI)) are widely distributed (i.e., endogenous) among non-methanotrophic bacteria and archaeal genomes (Orita et al., 2006).
[0082] As defined herein, the phrases recombinant host microorganism, genetically engineered host microorganism, engineered host microorganism and genetically modified host microorganism may be used interchangeably and refer to host microorganisms that have been genetically modified to (a) express one or more exogenous polynucleotides, (b) over-express one or more endogenous and/or one or more exogenous polynucleotides, such as those included in a vector, or which have an alteration in expression of an endogenous gene or (c) knock-out or down-regulate an endogenous gene. In addition, certain genes may be physically removed from the genome (e.g., knock-outs) or they may be engineered to have reduced, altered or enhanced activity.
[0083] The terms engineer, genetically engineer or genetically modify refer to any manipulation of a microorganism that results in a detectable change in the microorganism, wherein the manipulation includes, but is not limited to, introducing non-native metabolic functionality via heterologous (exogenous) polynucleotides or removing native-functionality via polynucleotide deletions, mutations or knock-outs. The term metabolically engineered generally involves rational pathway design and assembly of biosynthetic genes (or ORFs), genes associated with operons, and control elements of such polynucleotides, for the production of a desired metabolite. Metabolically engineered may further include optimization of metabolic flux by regulation and optimization of transcription, translation, protein stability and protein functionality using genetic engineering and appropriate culture condition including the reduction of, disruption, or knocking out of, a competing metabolic pathway that competes with an intermediate leading to a desired pathway.
[0084] As defined herein, the term introducing, as used in phrases such as introducing into the methanotroph host or introducing into the non-methanotroph host at least one polynucleotide open reading frame (ORF) or a gene thereof or a vector thereof includes methods known in the art for introducing polynucleotides into a cell, including, but not limited to transformation (e.g., calcium chloride, electroporation), transduction, transfection, conjugation and the like.
[0085] The phrases metabolically engineered microorganism and modified microorganism are used interchangeably herein, and refer not only to the particular subject host cell, but to the progeny or potential progeny of such a cell. Because certain modifications may occur in succeeding generations due to either mutation or environmental influences, such progeny may not, in fact, be identical to the parent cell, but are still included within the scope of the term as used herein.
[0086] The term mutation as used herein indicates any modification of a nucleic acid and/or polypeptide which results in an altered nucleic acid or polypeptide (i.e., relative to the wild-type nucleic acid or polypeptide sequence). Mutations include, for example, point mutations, substitutions, deletions, or insertions of single or multiple residues in a polynucleotide (or the encoded polypeptide), which includes alterations arising within a protein-encoding region of a gene as well as alterations in regions outside of a protein-encoding sequence, such as, but not limited to, regulatory or promoter sequences. A genetic alteration may be a mutation of any type. For instance, the mutation may constitute a point mutation, a frame-shift mutation, an insertion, or a deletion of part or all of a gene. In certain embodiments, a portion of a genetically modified microorganism's genome may be replaced with one or more heterologous (exogenous) polynucleotides. In some embodiments, the mutations are naturally-occurring. In other embodiments, the mutations are the results of artificial selection pressure. In still other embodiments, the mutations in the microorganism genome are the result of genetic engineering.
[0087] The term expression or expressed with respect to a gene sequence, an ORF sequence or polynucleotide sequence, refers to transcription of the gene, ORF or polynucleotide and, as appropriate, translation of the resulting mRNA transcript to a protein. Thus, as will be clear from the context, expression of a protein results from transcription and translation of the open reading frame sequence. The level of expression of a desired product in a host microorganism may be determined on the basis of either the amount of corresponding mRNA that is present in the host, or the amount of the desired product encoded by the selected sequence. For example, mRNA transcribed from a selected sequence can be quantitated by PCR or by northern hybridization (see Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, 1989). Protein encoded by a selected sequence can be quantitated by various methods (e.g., by ELISA, by assaying for the biological activity of the protein, or by employing assays that are independent of such activity, such as western blotting or radioimmunoassay, using antibodies that are recognize and bind reacting the protein).
[0088] The term endogenous, as used herein with reference to polynucleotides (and the polypeptides encoded therein), indicates polynucleotides and polypeptides that are expressed in the organism in which they originated (i.e., they are innate to the organism). In contrast, the terms heterologous and exogenous are used interchangeably, and as defined herein with reference to polynucleotides (and the polypeptides encoded therein), indicates polynucleotides and polypeptides that are expressed in an organism other than the organism from which they (i.e., the polynucleotide or polypeptide sequences) originated or where derived.
[0089] The term feedstock is defined as a raw material or mixture of raw materials supplied to a microorganism, or fermentation process, from which other products can be made. For example, as set forth in the present invention, a methane carbon source or a methanol carbon source or a formaldehyde carbon source, either alone or in combination, are feedstocks for a microorganism that produces a bio-fuel or bio-based chemical in a fermentation process. However, in addition to a feedstock (e.g., a methane substrate) of the invention, the fermentation media contains 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 pathways necessary for multi-carbon compound production.
[0090] The term substrate refers to any substance or compound that is converted, or meant to be converted, into another compound by the action of an enzyme. The term includes not only a single compound, but also combinations of compounds, such as solutions, mixtures and other materials which contain at least one substrate, or derivatives thereof. Further, the term substrate encompasses not only compounds that provide a carbon source suitable for use as a starting material (e.g., methane), but also intermediate and end product metabolites used in a pathway associated with a metabolically engineered microorganism as described herein.
[0091] The term fermentation or fermentation process is defined as a process in which a host microorganism is cultivated in a culture medium containing raw materials, such as feedstock and nutrients, wherein the microorganism converts raw materials, such as a feedstock, into products.
[0092] The term polynucleotide is used herein interchangeably with the term nucleic acid and refers to an organic polymer composed of two or more monomers including nucleotides, nucleosides or analogs thereof, including but not limited to single stranded or double stranded, sense or antisense deoxyribonucleic acid (DNA) of any length and, where appropriate, single stranded or double stranded, sense or antisense ribonucleic acid (RNA) of any length, including siRNA. The term nucleotide refers to any of several compounds that consist of a ribose or deoxyribose sugar joined to a purine or a pyrimidine base and to a phosphate group, and that are the basic structural units of nucleic acids. The term nucleoside refers to a compound (as guanosine or adenosine) that consists of a purine or pyrimidine base combined with deoxyribose or ribose and is found especially in nucleic acids. The term nucleotide analog or nucleoside analog refers, respectively, to a nucleotide or nucleoside in which one or more individual atoms have been replaced with a different atom or with a different functional group. Accordingly, the term polynucleotide includes nucleic acids of any length, including DNA, RNA, ORFs, analogs and fragments thereof.
[0093] As defined herein, the term open reading frame (hereinafter, ORF) means a nucleic acid or nucleic acid sequence (whether naturally occurring, non-naturally occurring, or synthetic) comprising an uninterrupted reading frame consisting of (i) an initiation codon, (ii) a series of two (2) of more codons representing amino acids, and (iii) a termination codon, the ORF being read (or translated) in the 5 to 3 direction.
[0094] It is understood that the polynucleotides described herein include genes and that the nucleic acid molecules described herein include vectors or plasmids. Accordingly, the term gene, refers to a polynucleotide that codes for a particular sequence of amino acids, which comprise all or part of one or more proteins or enzymes, and may include regulatory (non-transcribed) DNA sequences, such as promoter sequences, which determine for example the conditions under which the gene is expressed. The transcribed region of the gene may include untranslated regions, including introns, 5-untranslated region (UTR), and 3-UTR, as well as the coding sequence.
[0095] The term promoter refers to a nucleic acid 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 nucleic acid 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.
[0096] 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.
[0097] The term codon-optimized as it refers to genes or coding regions of nucleic acid molecules (or ORFs) 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.
[0098] The term operon refers to two or more genes which are transcribed as a single transcriptional unit from a common promoter. In certain embodiments, the genes, polynucleotides or ORFs comprising the operon are contiguous genes. It is understood that transcription of an entire operon can be modified (i.e., increased, decreased, or eliminated) by modifying the common promoter. Alternatively, any gene, polynucleotide or ORF, or any combination thereof in an operon can be modified to alter the function or activity of the encoded polypeptide. The modification can result in an increase or a decrease in the activity or function of the encoded polypeptide. Further, the modification can impart new activities on the encoded polypeptide.
[0099] A vector is any means by which a nucleic acid can be propagated and/or transferred between organisms, cells, or cellular components. Vectors include viruses, bacteriophage, pro-viruses, plasmids, phagemids, transposons, and artificial chromosomes such as YACs (yeast artificial chromosomes), BACs (bacterial artificial chromosomes), and PLACs (plant artificial chromosomes), and the like, that are episomes, that is, that replicate autonomously or can integrate into a chromosome of a host microorganism. A vector can also be a naked RNA polynucleotide, a naked DNA polynucleotide, a polynucleotide composed of both DNA and RNA within the same strand, a poly-lysine-conjugated DNA or RNA, a peptide-conjugated DNA or RNA, a liposome-conjugated DNA, or the like, that are not episomal in nature, or it can be an organism which comprises one or more of the above polynucleotide constructs such as an agrobacterium or a bacterium.
[0100] The term homolog, as used with respect to an original enzyme, polypeptide, gene or polynucleotide (or ORF encoding the same) of a first family or species, refers to distinct enzymes, genes or polynucleotides of a second family or species, which are determined by functional, structural or genomic analyses to be an enzyme, gene or polynucleotide of the second family or species, which corresponds to the original enzyme or gene of the first family or species. Most often, homologs will have functional, structural or genomic similarities. Techniques are known by which homologs of an enzyme, gene or polynucleotide can readily be cloned using genetic probes and PCR. Identity of cloned sequences as homologs can be confirmed using functional assays and/or by genomic mapping of the genes.
[0101] A polypeptide (or protein or enzyme) has homology or is homologous to a second polypeptide if the nucleic acid sequence that encodes the polypeptide has a similar sequence to the nucleic acid sequence that encodes the second polypeptide. Alternatively, a polypeptide has homology to a second polypeptide if the two proteins have similar amino acid sequences. Thus, the terms homologous proteins or homologous polypeptides is defined to mean that the two polypeptides have similar amino acid sequences. In certain embodiments of the invention, polynucleotides and polypeptides homologous to one or more polynucleotides and/or polypeptides set forth in Table 1 may be readily identified using methods known in the art for sequence analysis and comparison.
[0102] A homologous polynucleotide or polypeptide sequence of the invention may also be determined or identified by BLAST analysis (Basic Local Alignment Search Tool) or similar bioinformatic tools, which compare a query nucleotide or polypeptide sequence to a database of known sequences. For example, a search analysis may be done using BLAST to determine sequence identity or similarity to previously published sequences, and if the sequence has not yet been published, can give relevant insight into the function of the DNA or protein sequence.
Bioconversion of Methane to Multi-Carbon Compounds
[0103] In general, the conversion of methane (CH.sub.4) to multi-carbon compounds such as isobutanol ((CH.sub.3).sub.2CHCH.sub.2OH), 1-butanol or n-butanol (CH.sub.3CH.sub.2CH.sub.2CH.sub.2OH), ethanol (CH.sub.3CH.sub.2OH), fatty alcohols, fatty acid esters, 2,3-butanediol and the like, using a methanotrophic host microorganism, requires at least the following three steps, all of which are innate (or endogenous) with respect to methanotrophic organisms: (1) a methane (CH.sub.4) substrate is oxidized to methanol (CH.sub.3OH) via a methane monooxygenase (MMO) (e.g., particulate methane monooxygenase (pMMO) or soluble methane monooxygenase (sMMO)), (2) the methanol (CH.sub.3OH) is oxidized to formaldehyde (H.sub.2CO) via methanol dehydrogenase (MDH) and (3) the formaldehyde (H.sub.2CO) produced in step (2) above is assimilated into a central metabolism pathway (e.g., see type I (RuMP) and type II (serine) pathways described below).
[0104] In certain embodiments of the invention, a host microorganism is a methanotroph, which endogenously expresses a methane monooxygenase (MMO) enzyme and a methanol dehydrogenase (MDH) enzyme. In other embodiments of the invention, a host microorganism of the invention is a non-methanotrophic prokaryotic microorganism (e.g., a non-methanotrophic bacteria or archaea) or a eukaryotic microorganism (e.g., fungi and algae) engineered to utilize a methane substrate (as sole carbon source) for growth and energy. Thus, in certain embodiments of the invention, a non-methanotrophic microorganism is engineered to express (or over-express) an exogenous methane monooxygenase (MMO), an enzyme requisite to metabolize methane to methanol. The non-methanotroph host microorganisms of the invention comprise an endogenous dehydrogenase (MDH) enzyme, which converts methanol to formaldehyde. However, in certain embodiments, the non-methanotroph microorganism is further engineered to express an exogenous methanol dehydrogenase (MDH) enzyme, which converts methanol to formaldehyde. The expression of the exogenous MDH enzyme in a non-methanotroph host is not a strict requirement for the utilization of the methane substrate, but it is contemplated in certain embodiments, that the introduction and expression of an exogenous MDH in a non-methanotroph host thereof may facilitate, under certain growth conditions, the production of one or more multi-carbon compounds of the invention.
[0105] As mentioned briefly above with regard to methanotrophic host organisms, there are at least two known pathways (i.e., the ribulose monophosphate (RuMP) pathway and the serine pathway; Hanson & Hanson, 1996) for the assimilation of formaldehyde into central metabolism. In the Type I methanotroph RuMP pathway, formaldehyde combines with ribulose-5-phosphate to form hexulose-6-phosphate (catalyzed via hexulose-6-phosphate synthase), the hexulose-6-phosphate is then isomerized to fructose-6-phosphate (catalyzed via hexulose phosphate isomerase), which is an intermediate of a central metabolic pathway (i.e., glycolysis pathway). In the type II methanotroph serine pathway, formaldehyde reacts with tetrahydrofolate (THF) to form methylene-THF, the methylene-THF is then transferred to L-glycine to form L-serine, and finally the L-serine is transferred to glyoxylate to form hydroxypyruvate. The hydroxypyruvate formed is subsequently converted to 2-phosphoglycerate (catalyzed via hydroxypruvate reductase), which is an central metabolism intermediate of the glycolytic pathway.
[0106] Likewise, as mentioned briefly above, an endogenous pathway, which functions similarly (or analogous) to the ribulose monophosphate (RuMP) pathway in methanotrophs is also present in non-methanotrophic prokaryotes (Orita et al., 2006), wherein formaldehyde is fixed with ribulose 5-phosphate to form hexulose-6-phosphate (catalyzed via hexulose-6-phosphate synthase (HPS)) and then isomerized to fructose-6-phosphate (catalyzed via hexulose phosphate isomerase (PHI)), which is an intermediate of a central metabolic pathway. Thus, in certain preferred embodiments, a non-methanotrophic host microorganism of the invention comprises an endogenous RuMP pathway or an endogenous pathway analogous to the RuMP pathway. As defined herein, a pathway analogous to the RuMP pathway comprises at least a gene, polynucleotide or ORF encoding an enzyme having hexulose-6-phosphate synthase (HPS) activity from enzyme class EC 4.1.2.43 and at least a gene, polynucleotide or ORF encoding a an enzyme having hexulose phosphate isomerase (PHI) activity from enzyme class 5.3.1.27.
[0107] In other embodiments, wherein a non-methanotrophic host microorganism genome does not encode endogenous enzymes having HPS and PHI activity, the non-methanotroph host microorganism is genetically modified to express HPS and PHI enzymes. Thus, in certain embodiments, a gene, polynucleotide or ORF encoding a hexulose-6-phosphate synthase (HPS) is provided, wherein the gene, polynucleotide or ORF encodes a HPS polypeptide of enzyme class EC 4.1.2.43. In other embodiments, a gene, polynucleotide or ORF encoding a hexulose-6-phosphate synthase (HPS) is provided, wherein the gene, polynucleotide or ORF encodes a HPS polypeptide having at least 90% sequence homology to a M. capsulatus (Bath) HPS polypeptide of SEQ ID NO:173. In other embodiments, a gene, polynucleotide or ORF encoding a hexulose phosphate isomerase (PHI) is provided, wherein the gene, polynucleotide or ORF encodes a PHI polypeptide of enzyme class EC 5.3.1.27. In other embodiments, a gene, polynucleotide or ORF encoding a hexulose phosphate isomerase (PHI) is provided, wherein the gene, polynucleotide or ORF encodes a M. capsulatus (Bath) PHI polypeptide having at least 90% sequence homology to a PHI (also referred to as a sugar isomerase (SIS) domain) polypeptide of SEQ ID NO:175.
[0108] Once the formaldehyde has been assimilated into a central metabolic pathway of the methanotroph or non-methanotroph host organism (as described above), the fourth and final step for producing multi-carbon compounds from a methane substrate as described in steps (1)-(3) above, is the introduction of one or more nucleic acids into the host microorganism, wherein the one or more nucleic acids introduced encode one or more enzymes of a relevant multi-carbon compound pathway. Independent of the compound to be produced according to the present invention (e.g., isobutanol, 1-butanol, ethanol, fatty alcohols, fatty acid methyl esters, 2,3-butanediol and the like), any multi-carbon pathway introduced into a host microorganism must utilize a central metabolic molecule (e.g., pyruvate, acetyl-CoA, methionine and oxobutyrate) previously assimilated and introduced into the metabolic pathway through steps (1)-(3) described above. Stated another way, a salient feature of the present invention is the ability of the host microorganism to utilize methane (as a sole carbon source for growth and energy) and to produce multi-carbon compounds (via engineered metabolic pathways introduced therein), without the need for additional or supplemental carbon sources such as carbohydrates.
[0109] As defined herein, a relevant multi-carbon compound pathway, includes, but is not limited to, a 1-butanol pathway (which includes, but is not limited to, a fermentative 1-butanol pathway, a thiobutanoate pathway, a ketoacid pathway and a methylmalate pathway), an isobutanol pathway, a fatty alcohol pathway, a fatty acid methyl ester pathway and a 2,3-butanediol pathway. A multi-carbon compound pathway as further defined herein, may include one specific enzyme from the pathway, multiple enzymes from the pathway or all of the enzymes of the pathway. It will be understood by a person of skill in the art, that the selection of one or more specific pathway enzymes (and nucleic acids encoding the same) may be dependent on the host microorganism (e.g., certain methanotroph hosts or non-methanotroph hosts may endogenously encode and express one or more enzymes of a given pathway).
[0110] For example,
[0111] As depicted in
[0112] Thus, in certain embodiments, the present invention is directed to a method for producing isobutanol from a methane substrate comprising the steps of (a) providing a methanotrophic host microorganism that metabolizes methane (CH.sub.4) to methanol (CH.sub.3OH) and methanol to formaldehyde (H.sub.2CO); (b) introducing into the methanotroph host and expressing at least one polynucleotide open reading frame (ORF), under the control of suitable regulatory sequences, wherein the at least one polynucleotide ORF encodes a polypeptide that catalyzes a reaction in an isobutanol pathway; (c) feeding the methanotroph host of step (b) a methane substrate under suitable growth conditions, wherein the host metabolizes methane to formaldehyde as set forth in step (a), wherein the formaldehyde is converted to pyruvate by means of an endogenous RuMP pathway or a serine pathway and the host metabolizes pyruvate to produce isobutanol, and (d) optionally recovering the isobutanol produced.
[0113] In one particular embodiment, the one or more polynucleotide ORFs introduced in step (b) encode an isobutanol pathway polypeptide thereof selected from an Enzyme Class (EC) comprising EC 2.2.1.6, EC 1.1.1.86, EC 4.2.1.9, EC 4.1.1.72 and EC 1.1.1.1. In other embodiments, the one or more polynucleotide ORFs introduced in step (b) encode an isobutanol pathway polypeptide selected from the group consisting of acetolactate synthase (ALS), ketol-acid reductoisomerase (KARI), dihydroxy-acid dehydratase (DHAD), ketoacid decarboxylase (KDC) and alcohol dehydrogenase (ADH). In certain embodiments, the ALS polypeptide catalyzes the substrate to product conversion of pyruvate to acetolactate; the KARI polypeptide catalyzes the substrate to product conversion of acetolactate to 2,3-dihydroxyisovalerate; the DHAD polypeptide catalyzes the substrate to product conversion of 2,3-dihydroxyisovalerate to ketoisovalerate; the KDC polypeptide catalyzes the substrate to product conversion of ketoisovalerate to isobutryaldehyde and ADH polypeptide catalyzes the substrate to product conversion of isobutyraldehyde to isobutanol. In other embodiments, the ALS polypeptide comprises an amino acid sequence comprising at least 90% sequence homology to SEQ ID NO:2, the KARI polypeptide comprises an amino acid sequence comprising at least 90% sequence homology to SEQ ID NO:4, the DHAD polypeptide comprises an amino acid sequence comprising at least 90% sequence homology to SEQ ID NO:6, the KDC polypeptide comprises an amino acid sequence comprising at least 90% sequence homology to SEQ ID NO: 8 and the ADH polypeptide comprises an amino acid sequence comprising at least 90% sequence homology to SEQ ID NO:10. In yet other embodiments, the one or more polynucleotide ORFs introduced in step (b) encode the complete isobutanol pathway comprising an ALS polypeptide, a KARI polypeptide, a DHAD polypeptide, a KDC polypeptide and an ADH polypeptide. In certain embodiments, the ORFs encoding the complete isobutanol pathway are comprised in one operon, two operons or three operons, wherein each operon may comprise the same promoter or a different promoter, wherein the same or different promoters may be constitutive or inducible.
[0114] In certain embodiments, a methanotroph host microorganism is modified or genetically engineered to express one or more enzymes of a metabolic pathway capable of producing n-butanol, isobutanol, fatty (or aliphatic long chain) alcohols, fatty acid methyl esters and the like. In particular embodiments, a methanotroph of the invention is selected from genera consisting of Methylobacter, Methylomicrobium, Methylomonas, Methylocaldum, Methylococcus, Methylosoma, Methylosarcina, Methylothermus, Methylohalobius, Methylogaea, Methylovulum, Crenothrix, Clonothrix, Methylosphaera, Methylocapsa, Methylocella, Methylosinus, Methylocystis and Methyloacidophilum. In other embodiments, the methanotroph is from the phylum Verrucomicrobia. Previously published work has shown that several species within these taxa can be genetically transformed by introducing DNA constructs on plasmid vectors (Stafford et al., 2003), or by integrating them into the bacterial chromosome (Welander & Summons, 2012). Thus, a vector construct of the invention will typically comprise the pathway genes or polynucleotide ORFs, which are initially constructed and cloned into E. coli to generate sufficient quantities of the vector, and then the vectors are subsequently transformed into the host microorganism for expression.
[0115] In other embodiments, the invention is directed to a method for producing isobutanol from a methane substrate comprising the steps of (a) providing a non-methanotroph host microorganism which has been genetically engineered to express a methane monooxygenase (MMO) (and optionally a methanol dehydrogenase (MDH)) and wherein the non-methanotroph host comprises either an endogenous RuMP pathway or an endogenous serine pathway, (b) introducing into the host and expressing at least one polynucleotide open reading frame (ORF), under the control of suitable regulatory sequences, wherein the at least one polynucleotide ORF encodes a polypeptide that catalyzes a reaction in an isobutanol pathway; (c) feeding the host of step (b) a methane substrate under suitable growth conditions, wherein the MMO polypeptide catalyzes the substrate to product conversion of methane to methanol, an endogenous MDH polypeptide catalyzes the substrate to product conversion of methanol to formaldehyde, the formaldehyde produced is converted to pyruvate through an endogenous RuMP or serine pathway and the host metabolizes pyruvate to produce isobutanol, and (d) optionally recovering the isobutanol produced. Methods for heterologous expression of pMMO genes have been described in Gou et al. (2006). Methods for heterologous expression of sMMO genes have been described in Lloyd et al. (1999). Suitable microbial hosts for heterologous expression include microorganisms that have the ability to process methanol and formaldehyde, that have multiple heterotrophic growth modes, and/or that can assemble complex membranes and metalloprotein complexes. Such organisms include methylotrophic yeasts (e.g., Pichia pastoris) as well as bacteria such as Pseudomonas putida, Cupriavidus metallidurans and Rhodobacter sphaeroides.
[0116] In certain embodiments, the one or more polynucleotide ORFs introduced in step (b) above, encode an isobutanol pathway polypeptide selected from an Enzyme Class (EC) comprising EC 2.2.1.6, EC 1.1.1.86, EC 4.2.1.9, EC 4.1.1.72 and EC 1.1.1.1. In other embodiments, the one or more polynucleotide ORFs introduced in step (b) encode an isobutanol pathway polypeptide selected from the group consisting of acetolactate synthase (ALS), ketol-acid reductoisomerase (KARI), dihydroxy-acid dehydratase (DHAD), ketoacid decarboxylase (KDC) and alcohol dehydrogenase (ADH). In yet other embodiments, the ALS polypeptide catalyzes the substrate to product conversion of pyruvate to acetolactate; the KARI polypeptide catalyzes the substrate to product conversion of acetolactate to 2,3-dihydroxyisovalerate; the DHAD polypeptide catalyzes the substrate to product conversion of 2,3-dihydroxyisovalerate to ketoisovalerate; the KDC polypeptide catalyzes the substrate to product conversion of ketoisovalerate to isobutryaldehyde and ADH polypeptide catalyzes the substrate to product conversion of isobutyraldehyde to isobutanol.
[0117] In one particular embodiment, the ALS polypeptide comprises an amino acid sequence comprising at least 90% sequence homology to SEQ ID NO:2, the KARI polypeptide comprises an amino acid sequence comprising at least 90% sequence homology to SEQ ID NO:4, the DHAD polypeptide comprises an amino acid sequence comprising at least 90% sequence homology to SEQ ID NO:6, the KDC polypeptide comprises an amino acid sequence comprising at least 90% sequence homology to SEQ ID NO: 8 and the ADH polypeptide comprises an amino acid sequence comprising at least 90% sequence homology to SEQ ID NO:10. In certain other embodiments, the one or more polynucleotide ORFs introduced in step (b) encode the complete isobutanol pathway comprising an ALS polypeptide, a KARI polypeptide, a DHAD polypeptide, a KDC polypeptide and an ADH polypeptide. In another embodiment, the methane monooxygenase (MMO) is a soluble MMO of Enzyme Class EC 1.14.13.25 or a particulate MMO of Enzyme Class 1.14.18.3. In other embodiments, the MMO comprises an amino acid sequence having at least 90% sequence homology to a particulate methane monooxygenase (pMMO) of operon 1 comprising pmoC1 subunit 1 (SEQ ID NO:12), pmoA subunit 1 (SEQ ID NO:14), pmoB subunit 1 (SEQ ID NO: 16) or a pMMO of operon 2 comprising pmoC subunit 2 (SEQ ID NO:18), pmoA subunit 2 (SEQ ID NO:20), pmoB subunit 2 (SEQ ID NO:22). In other embodiments, the MMO comprises an amino acid sequence having at least 90% sequence homology to a soluble methane monooxygenase (sMMO) selected from mmoX (SEQ ID NO:24), mmoY (SEQ ID NO:26), mmoB (SEQ ID NO: 28), mmoZ (SEQ ID NO:30), mmoD (SEQ ID NO:32) or mmoC (SEQ ID NO:34).
[0118] In certain embodiments where an exogenous methanol dehydrogenase (MDH) is optionally provided and expressed in a host microorganism, the MDH is a polypeptide from Enzyme Class 1.14.18.3. In certain other embodiments, the MDH comprises an amino acid sequence comprising at least 90% sequence homology to mxaF (SEQ ID NO:36), mxaJ (SEQ ID NO:38), mxaG (SEQ ID NO:40), mxal (SEQ ID NO:42), mxaR (SEQ ID NO:44), mxaA (SEQ ID NO:46), mxaC (SEQ ID NO:48), mxaK (SEQ ID NO:50), mxaL (SEQ ID NO: 52) or mcaD (SEQ ID NO:54).
[0119] In other embodiments, the invention is directed to a method for producing 1-butanol from a methane substrate comprising the steps of (a) providing a methanotrophic host microorganism that metabolizes methane (CH.sub.4) to methanol (CH.sub.3OH) and methanol to formaldehyde (H.sub.2CO), (b) introducing into the methanotroph host and expressing at least one polynucleotide open reading frame (ORF), under the control of suitable regulatory sequences, wherein the at least one polynucleotide ORF encodes a polypeptide that catalyzes a reaction in a 1-butanol pathway; (c) feeding the methanotroph host of step (b) a methane substrate under suitable growth conditions, wherein the host metabolizes methane to formaldehyde as set forth in step (a), wherein the formaldehyde is converted to pyruvate by means of an endogenous type I RuMP pathway or a type II serine pathway and the host metabolizes pyruvate to produce 1-butanol, and (d) optionally recovering the 1-butanol produced. In certain embodiments, the one or more polynucleotide ORFs introduced in step (b) encode a 1-butanol pathway polypeptide selected from an Enzyme Class (EC) comprising EC 4.3.1.19, EC 2.3.3.6, EC 4.2.1.33, EC 1.1.1.85, EC 4.1.1.72, and EC 1.1.1.1. In another embodiment, the one or more polynucleotide ORFs introduced in step (b) encode a 1-butanol pathway polypeptide selected from the group consisting of L-threonine ammonia-lyase, 2-ethylmalate synthase (or 2-isopropylmalate synthase), isopropylmalate isomerase (or 3-isopropylmalate dehydratase), 3-isopropylmalate dehydrogenase, 2-ketoacid decarboxylase (KDC) and alcohol dehydrogenase (ADH). In certain other embodiments, L-threonine ammonia-lyase catalyzes the substrate to product conversion of L-threonine to 2-oxybutanoate (2-ketobutyrate) and ammonia; the 2-ethylmalate synthase catalyzes the substrate to product conversion of 2-oxybutanoate and acetyl-CoA to 2-ethylmalate; the isopropylmalate isomerase catalyzes the substrate to product conversion of 2-ethylmalate to 3-ethylmalate; the 3-isopropylmalate dehydrogenase catalyzes the substrate to product conversion of 3-ethylmalate to 2-ketovalerate, CO.sub.2 and NADH; the KDC catalyzes the substrate to product conversion of 2-ketovalerate to butryaldehyde and the ADH catalyzes the substrate to product conversion of butyraldehyde to 1-butanol.
[0120] In certain embodiments, a L-threonine ammonia-lyase comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO:56, a 2-ethylmalate synthase comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO:58, a isopropylmalate isomerase comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO:60 and SEQ ID NO:62, a 3-isopropylmalate dehydrogenase comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO:64, the KDC comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO:8 and the ADH comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO: 10. In one particular embodiment, the one or more polynucleotide ORFs introduced in step (b) encode the complete 1-butanol pathway comprising an L-threonine ammonia-lyase, a 2-ethylmalate synthase, an isopropylmalate isomerase, a 3-isopropylmalate dehydrogenase, a KDC and an ADH.
[0121] In other embodiments, the invention is directed to a method for producing 1-butanol from a methane substrate comprising the steps of (a) providing a non-methanotroph host microorganism which has been genetically engineered to express a methane monooxygenase (MMO) (and optionally a methanol dehydrogenase (MDH)) and wherein the non-methanotroph host comprises either an endogenous RuMP pathway or an endogenous serine pathway, (b) introducing into the host and expressing at least one polynucleotide open reading frame (ORF), under the control of suitable regulatory sequences, wherein the at least one polynucleotide ORF encodes a polypeptide that catalyzes a reaction in a 1-butanol pathway; (c) feeding the host of step (b) a methane substrate under suitable growth conditions, wherein the MMO polypeptide catalyzes the substrate to product conversion of methane to methanol, an endogenous MDH polypeptide catalyzes the substrate to product conversion of methanol to formaldehyde, the formaldehyde produced is converted to pyruvate through an endogenous RuMP or serine pathway and the host metabolizes pyruvate to produce 1-butanol, and (d) optionally recovering the 1-butanol produced.
[0122] In certain embodiments, the non-methanotroph host microorganism is genetically modified to express an exogenous methane monooxygenase (MMO). In one embodiment, the methane monooxygenase is a soluble MMO (sMMO) of Enzyme Class EC 1.14.13.25 or a particulate MMO (pMMO) of Enzyme Class 1.14.18.3. In other embodiments, the MMO comprises an amino acid sequence having at least 90% sequence homology to a particulate methane monooxygenase (pMMO) of operon 1 comprising pmoC1 subunit 1 (SEQ ID NO: 12), pmoA subunit 1 (SEQ ID NO:14), pmoB subunit 1 (SEQ ID NO: 16) or a pMMO of operon 2 comprising pmoC subunit 2 (SEQ ID NO: 18), pmoA subunit 2 (SEQ ID NO: 20), pmoB subunit 2 (SEQ ID NO:22). In other embodiments, the MMO comprises an amino acid sequence having at least 90% sequence homology to a soluble methane monooxygenase (sMMO) selected from mmoX (SEQ ID NO:24), mmoY (SEQ ID NO:26), mmoB (SEQ ID NO:28), mmoZ (SEQ ID NO:30), mmoD (SEQ ID NO:32) or mmoC (SEQ ID NO: 34).
[0123] In certain embodiments where an exogenous methanol dehydrogenase (MDH) is optionally provided and expressed in a host microorganism, the MDH is a polypeptide from Enzyme Class 1.14.18.3. In certain other embodiments, the MDH comprises an amino acid sequence comprising at least 90% sequence homology to mxaF (SEQ ID NO:36), mxaJ (SEQ ID NO:38), mxaG (SEQ ID NO:40), mxal (SEQ ID NO:42), mxaR (SEQ ID NO:44), mxaA (SEQ ID NO:46), mxaC (SEQ ID NO:48), mxaK (SEQ ID NO:50), mxaL (SEQ ID NO: 52) or mcaD (SEQ ID NO:54).
[0124] In one particular embodiment, the one or more polynucleotide ORFs introduced in step (b) encode a 1-butanol pathway polypeptide selected from an Enzyme Class (EC) comprising EC 4.3.1.19, EC 2.3.3.6, EC 4.2.1.33, EC 1.1.1.85, EC 4.1.1.72, and EC 1.1.1.1. In another embodiment, the one or more polynucleotide ORFs introduced in step (b) encode a 1-butanol pathway polypeptide selected from the group consisting of L-threonine ammonia-lyase, 2-ethylmalate synthase (or 2-isopropylmalate synthase), isopropylmalate isomerase (or 3-isopropylmalate dehydratase), 3-isopropylmalate dehydrogenase, 2-ketoacid decarboxylase (KDC) and alcohol dehydrogenase (ADH). In certain other embodiments, L-threonine ammonia-lyase catalyzes the substrate to product conversion of L-threonine to 2-oxybutanoate (2-ketobutyrate) and ammonia; the 2-ethylmalate synthase catalyzes the substrate to product conversion of 2-oxybutanoate and acetyl-CoA to 2-ethylmalate; the isopropylmalate isomerase catalyzes the substrate to product conversion of 2-ethylmalate to 3-ethylmalate; the 3-isopropylmalate dehydrogenase catalyzes the substrate to product conversion of 3-ethylmalate to 2-ketovalerate, CO.sub.2 and NADH; the KDC catalyzes the substrate to product conversion of 2-ketovalerate to butryaldehyde and the ADH catalyzes the substrate to product conversion of butyraldehyde to 1-butanol.
[0125] In certain embodiments, a L-threonine ammonia-lyase comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO:56, a 2-ethylmalate synthase comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO:58, a isopropylmalate isomerase comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO:60 and SEQ ID NO:62, a 3-isopropylmalate dehydrogenase comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO:64, the KDC comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO:8 and the ADH comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO:10. In one particular embodiment, the one or more polynucleotide ORFs introduced in step (b) encode the complete 1-butanol pathway comprising an L-threonine ammonia-lyase, a 2-ethylmalate synthase, an isopropylmalate isomerase, a 3-isopropylmalate dehydrogenase, a KDC and an ADH.
[0126] In certain other embodiments, the invention is directed to a method for producing fatty alcohols from a methane substrate comprising the steps of (a) providing a methanotrophic host microorganism that metabolizes methane (CH.sub.4) to methanol (CH.sub.3OH) and methanol to formaldehyde (H.sub.2CO), (b) introducing into the methanotroph host and expressing a polynucleotide open reading frame (ORF), under the control of suitable regulatory sequences, wherein the polynucleotide ORF encodes a fatty-acyl-CoA reductase (FAR); (c) feeding the methanotroph host of step (b) a methane substrate under suitable growth conditions, wherein the host metabolizes methane to formaldehyde as set forth in step (a), wherein the formaldehyde is converted to acetyl-CoA by means of an endogenous type I RuMP pathway or a type II serine pathway and the host metabolizes acetyl-CoA to produce a fatty alcohol, and (d) recovering the fatty alcohol produced. In certain embodiments, the FAR polypeptide is further defined as a polypeptide from Enzyme Class EC 1.2.1.50. In yet other embodiments, the FAR polypeptide catalyzes the substrate to product conversion of fatty acetyl-CoA to a fatty alcohol. In another embodiment, a FAR polypeptide comprises an amino acid sequence having at least 90% sequence homology to SEQ ID NO:66.
[0127] In still other embodiments, the invention is directed to a method for producing a fatty alcohol from a methane substrate comprising the steps of (a) providing a non-methanotroph host microorganism which has been genetically engineered to express a methane monooxygenase (MMO) (and optionally a methanol dehydrogenase (MDH)) and wherein the non-methanotroph host comprises either an endogenous RuMP pathway or an endogenous serine pathway, (b) introducing into the host microorganism and expressing a polynucleotide open reading frame (ORF), under the control of suitable regulatory sequences, wherein the polynucleotide ORF encodes a fatty-acyl-CoA reductase (FAR), (c) feeding the host microorganism of step (b) a methane substrate under suitable growth conditions, wherein the host metabolizes methane to formaldehyde as set forth in step (a), wherein the formaldehyde is converted to acetyl-CoA by means of an endogenous type I RuMP pathway or a type II serine pathway and the host metabolizes acetyl-CoA to produce a fatty alcohol, and (d) optionally recovering the fatty alcohol produced.
[0128] In certain embodiments, the non-methanotroph host microorganism is genetically modified to express an exogenous methane monooxygenase (MMO). In one embodiment, the methane monooxygenase is a soluble MMO (sMMO) of Enzyme Class EC 1.14.13.25 or a particulate MMO (pMMO) of Enzyme Class 1.14.18.3. In other embodiments, the MMO comprises an amino acid sequence having at least 90% sequence homology to a particulate methane monooxygenase (pMMO) of operon 1 comprising pmoC1 subunit 1 (SEQ ID NO:12), pmoA subunit 1 (SEQ ID NO:14), pmoB subunit 1 (SEQ ID NO: 16) or a pMMO of operon 2 comprising pmoC subunit 2 (SEQ ID NO:18), pmoA subunit 2 (SEQ ID NO: 20), pmoB subunit 2 (SEQ ID NO:22). In other embodiments, the MMO comprises an amino acid sequence having at least 90% sequence homology to a soluble methane monooxygenase (sMMO) selected from mmoX (SEQ ID NO:24), mmoY (SEQ ID NO:26), mmoB (SEQ ID NO:28), mmoZ (SEQ ID NO:30), mmoD (SEQ ID NO:32) or mmoC (SEQ ID NO: 34).
[0129] In certain embodiments, where an exogenous methanol dehydrogenase (MDH) is optionally provided and expressed in a host microorganism, the MDH is a polypeptide from Enzyme Class 1.14.18.3. In certain other embodiments, the MDH comprises an amino acid sequence comprising at least 90% sequence homology to mxaF (SEQ ID NO:36), mxaJ (SEQ ID NO:38), mxaG (SEQ ID NO:40), mxal (SEQ ID NO:42), mxaR (SEQ ID NO:44), mxaA (SEQ ID NO:46), mxaC (SEQ ID NO:48), mxaK (SEQ ID NO:50), mxaL (SEQ ID NO: 52) or mcaD (SEQ ID NO:54).
[0130] In another embodiment, the invention is directed to a method for producing a fatty acid ester from a methane substrate comprising the steps of (a) providing a methanotrophic host microorganism that metabolizes methane (CH.sub.4) to methanol (CH.sub.3OH) and methanol to formaldehyde (H.sub.2CO), (b) introducing into the methanotroph host and expressing a polynucleotide open reading frame (ORF), under the control of suitable regulatory sequences, wherein the polynucleotide ORF encodes a wax ester synthase (WES); (c) feeding the methanotroph host of step (b) a methane substrate under suitable growth conditions, wherein the host metabolizes methane to formaldehyde as set forth in step (a), wherein the formaldehyde is converted to acetyl-CoA by means of an endogenous type I RuMP pathway or a type II serine pathway and the host metabolizes fatty-acyl-CoA and alcohols to produce a fatty acid ester, and (d) recovering the fatty acid ester produced. In one particular embodiment, the WES polypeptide is further defined as a polypeptide from Enzyme Class EC 2.3.1.75. In another embodiment, the WES polypeptide catalyzes the substrate to product conversion of a fatty acid to a fatty acid esters. In another embodiment, the WES polypeptide catalyzes the substrate to product conversion of fatty alcohol and acyl-CoA to fatty acid esters. In one particular embodiment, the WES polypeptide comprises an amino acid sequence having at least 90% sequence homology to a WES polypeptide selected from SEQ ID NO:68, SEQ ID NO: 70, SEQ ID NO: 72, SEQ ID NO: 74, SEQ ID NO: 76 and SEQ ID NO: 78.
[0131] In another embodiment, the invention is directed to a method for producing a fatty acid ester from a methane substrate comprising the steps of (a) providing a non-methanotroph host microorganism which has been genetically engineered to express a methane monooxygenase (MMO) (and optionally a methanol dehydrogenase (MDH)) and wherein the non-methanotroph host comprises either an endogenous RuMP pathway or an endogenous serine pathway, (b) introducing into the host microorganism and expressing at least one polynucleotide open reading frame (ORF), under the control of suitable regulatory sequences, wherein the at least one polynucleotide ORF encodes a wax ester synthase; (c) feeding the host of step (b) a methane substrate under suitable growth conditions, wherein the MMO polypeptide catalyzes the substrate to product conversion of methane to methanol, an endogenous MDH polypeptide catalyzes the substrate to product conversion of methanol to formaldehyde, wherein the formaldehyde is converted to acetyl-CoA by means of an endogenous RuMP or serine pathway and the host metabolizes fatty-acyl-CoA and alcohols to produce a fatty acid ester, and (d) recovering the fatty acid ester produced.
[0132] In one particular embodiment, the WES polypeptide is further defined as a polypeptide from Enzyme Class EC 2.3.1.75. In another embodiment, the WES polypeptide catalyzes the substrate to product conversion of a fatty acid to a fatty acid ester. In another embodiment, the WES polypeptide catalyzes the substrate to product conversion of fatty alcohol and acyl-CoA to fatty acid esters. In one particular embodiment, the WES polypeptide comprises an amino acid sequence having at least 90% sequence homology to a WES polypeptide selected from SEQ ID NO:68, SEQ ID NO: 70, SEQ ID NO: 72, SEQ ID NO: 74, SEQ ID NO: 76 and SEQ ID NO: 78.
[0133] In certain embodiments, the non-methanotroph host microorganism is genetically modified to express an exogenous methane monooxygenase (MMO). In one embodiment, the methane monooxygenase is a soluble MMO (sMMO) of Enzyme Class EC 1.14.13.25 or a particulate MMO (pMMO) of Enzyme Class 1.14.18.3. In other embodiments, the MMO comprises an amino acid sequence having at least 90% sequence homology to a particulate methane monooxygenase (pMMO) of operon 1 comprising pmoC1 subunit 1 (SEQ ID NO:12), pmoA subunit 1 (SEQ ID NO:14), pmoB subunit 1 (SEQ ID NO: 16) or a pMMO of operon 2 comprising pmoC subunit 2 (SEQ ID NO: 18), pmoA subunit 2 (SEQ ID NO: 20), pmoB subunit 2 (SEQ ID NO:22). In other embodiments, the MMO comprises an amino acid sequence having at least 90% sequence homology to a soluble methane monooxygenase (sMMO) selected from mmoX (SEQ ID NO:24), mmoY (SEQ ID NO:26), mmoB (SEQ ID NO:28), mmoZ (SEQ ID NO:30), mmoD (SEQ ID NO:32) or mmoC (SEQ ID NO: 34).
[0134] In certain embodiments, where an exogenous methanol dehydrogenase (MDH) is optionally provided and expressed in a host microorganism, the MDH is a polypeptide from Enzyme Class 1.14.18.3. In certain other embodiments, the MDH comprises an amino acid sequence comprising at least 90% sequence homology to mxaF (SEQ ID NO:36), mxaJ (SEQ ID NO:38), mxaG (SEQ ID NO:40), mxal (SEQ ID NO:42), mxaR (SEQ ID NO:44), mxaA (SEQ ID NO:46), mxaC (SEQ ID NO:48), mxaK (SEQ ID NO:50), mxaL (SEQ ID NO: 52) or mcaD (SEQ ID NO:54).
[0135] In certain other embodiments, the invention is directed to methods of producing 2,3-butanediol from a methane substrate. The compound 2,3-butanediol (a four-carbon diol) is an important intermediate for the chemical industry. At the commercial scale, 2,3-butanediol is mainly produced or generated from petroleum, where it serves as a precursor for the production of various commodity and specialty chemicals, such as the solvent methyl ethyl ketone (MEK), gamma-butyrolactone (GBL) and 1,3-butadiene. The biological production of 2,3-butanediol from methane requires engineering the native (or endogenous) metabolism of methanotrophs to take advantage of their endogenous production of (R)-acetoin (
[0136] General methods for gene synthesis and DNA cloning, as well as vector and plasmid construction, are well known in the art, and are described in a number of publications (Lipps, 2008; Peccoud, 2012; Ausubel et al., 2002). More specifically, techniques such as digestion and ligation-based cloning, as well as in vitro and in vivo recombination methods, can be used to assemble DNA fragments encoding a polypeptide that catalyzes a substrate to product conversion into a suitable vector. These methods include restriction digest cloning, sequence- and ligation-independent Cloning (SLIC) (Li & Elledge, 2012), Golden Gate cloning (Engler et al., 2009), Gibson assembly (Gibson et al., 2009), and the like (Merryman & Gibson, 2012; Wang et al., 2012). Some of these methods can be automated and miniaturized for high-throughput applications (Yehezkel et al., 2011; Ma et al., 2012).
[0137] In certain embodiments, the cloning procedures use in vitro homologous recombination, to insert DNA fragments into a vector (e.g., the In-Fusion kit from Clontech Laboratories, Inc. (Mountain View, CA)). For example, (1) the recipient vector is linearized by a restriction digest and purified; (2) PCR primers that are complementary to the fragment to be cloned and that are complementary (with 15-base pair extensions) to the ends of the linearized vector are used to amplify the insert, using high-fidelity polymerase; (3) the size of the PCR amplicon is verified by agarose gel electrophoresis; (4) the PCR product is purified by a spin-column; (5) the In-Fusion reaction is run according to the manufacturer's instructions; (6) competent E. coli cells are transformed with 2.5 L of the reaction products; (7) positive transformants are selected from colonies grown on antibiotic selection medium and transferred to individual liquid cultures with the appropriate antibiotic; (8) the cells are harvested after overnight growth at 37 C. with 200 rpm shaking and (9) the plasmid DNA is extracted and analyzed for the correct insert.
[0138] The plasmid vector is chosen so that it will be capable of replicating in both an E. coli host (for cloning and amplification) and a methanotrophic or non-methanotrophic host microorganism (for metabolic pathway expression). The plasmid can be transferred from the E. coli donor cell to the recipient cell via bacterial conjugation. In addition, the vector contains a promoter sequence upstream of the one or more polynucleotide ORFs that are to be expressed. The promoter sequence can be included as part of the insert so that it can be adjusted and tested for each new construct. Broad-host-range (bhr) vectors for different gram-negative bacterial hosts have been described in the literature (Marx & Lidstrom, 2001). These vectors typically contain the following components: (1) an origin of replication that is functional in E. coli (colE1); (2) an oriV/IncP origin of replication for the non-E. coli host; (3) an oriT/IncP origin of transfer, which is needed for transferring a bacterial plasmid from a bacterial host such as E. coli to the recipient during bacterial conjugation; (4) a traJ gene, which codes for a transcriptional activator that initiates production of the proteins needed for conjugative transfer; and (5) a trfA, the replication initiation protein gene of plasmid RK2 which binds to a activates oriV.
[0139] In one embodiment, the conjugative bhr plasmid is based on pCM132 (GenBank Accession No. AF327720, SEQ ID NO:79) (Marx & Lidstrom, 2001), which has been engineered to contain a kanamycin resistance gene for plasmid selection and a lacZ (beta-galactosidase) gene for identifying plasmids with DNA inserts based on colony color using indolyl-galactoside-based substrates. Genes (or polynucleotide ORFs thereof) of interest can be inserted into the polylinker region that lies between the rrnB transcription terminator and the 5-end of the lacZ gene (e.g., see,
[0140] Typical gene cassettes for expressing an engineered metabolic pathway in a host microorganism such as a methanotroph are shown in
[0141] In one embodiment, the plasmids generated as part of the present invention are based on the broad-host-range expression vector pCM132 (Marx & Lidstrom, 2001). In this embodiment, the use of the Clontech (catalog no. 639647) InFusion HD Cloning System kit is one example of how to construct plasmids, but is not meant to limit or exclude other methods that are known in the art, including Gibson assembly, yeast in vivo recombination, PCR Splicing by Overlap Extension, or any combination of these with standard molecular biology techniques.
[0142] In certain embodiments of the invention, the plasmids of interest are generated in a modular fashion such that various modules, including suitable regulatory sequences, can be easily assembled or replaced as needed and are amenable to scaled-up, high-throughput assembly. The plasmids are designed to consist of multiple linear modules: a vector backbone and one or more vector inserts. The 5 and 3 ends of individual modules have overlapping sequence homology to the ends of adjacent modules within the designed plasmid. The overlapping homology between the modules allows them to be assembled into a circular plasmid using the Clontech InFusion HD Cloning System kit or other assembly method known in the art. Primers were designed to introduce homologous ends to the PCR-amplified products to facilitate assembly.
[0143] Vector backbones of the invention contain the components of the plasmid that will remain constant. In certain embodiments, the broad-host range vector pCM132 is modified to produce vector backbones for the plasmids (vectors) of the invention. The pCM132 vector, further described below in the Examples section, consists of the following components: trrnB terminator, kanamycin resistance gene, trfA, IncP oriT, IncP oriV, colE1 ori, and lacZ. This parental vector was modified to replace lacZ with a vector insert that contains promoter sequence(s) to produce plasmids pJSvec (SEQ ID NO: 80) and pMZT3 (SEQ ID NO: 81). In certain embodiments of the invention, vector backbones were PCR-amplified with the NEB Phusion master mix (M0531L) according to the manufacturer's instructions, unless specified otherwise.
[0144] The general rationale or procedure for selecting the appropriate ORFs for a given pathway was to examine a list of pathway-relevant genes as specified in the literature. Using this set of pathway-relevant genes as a target, BLAST searches were run, looking for genes in three groups: (1) similar genes found in microbial hosts that are phylogenetically close to the ones already listed in the literature, (2) similar genes found in microbes that are phylogenetically distant from the microbial host of the targeted gene, and (3) homologs that are similar to the target gene but that are found in the wild-type methanotroph or non-methanotroph organism that is to be used as the expression host. An example of the above strategy would be to target the kivD gene (encoding alpha-ketoisovalerate decarboxylase) from Lactococcus lactis: the first group would contain genes from species similar to L. lactis, including Lactococcus itself; the second group would be genes similar to kivD, but found in organisms phylogenetically distant from L. lactis; and finally the last group would include a kivD gene in a microbe of interest, specifically, Methylococcus capsulatus (Bath). Thus, in certain embodiments of the invention, the exemplary polynucleotide and polypeptide sequences set forth in Table 1 are used to identify similar or homologous polynucleotide, genes, ORFs and polypeptides found in microbial hosts that are (1) phylogenetically close to the ones already listed, (2) found in microbes that are phylogenetically distant from the microbial host of the targeted sequence, and (3) homologs that are similar to the target gene but that are found in the wild-type methanotroph or non-methanotroph organism that is to be used as the expression host.
[0145] For example, genes encoding similar proteins or polypeptides to those of the invention may isolated directly by using all or a portion of a nucleic acid (e.g., see Table 1, below) or a primer sequence (e.g., see Table 2, below) as DNA hybridization probes to screen libraries from any desired microorgansim using methodology well known to those skilled in the art. Specific oligonucleotide probes based upon these nucleic acid sequences can be designed and synthesized by methods known in the art (Sambrook et al., 1989; Ausubel et al., 1987). Moreover, the entire sequence can be used directly to synthesize DNA probes by methods known to the skilled artisan such as random primers, DNA labeling, nick translation, or end-labeling techniques, or RNA probes using available in vitro transcription systems. In addition, specific primers can be designed and used to amplify a part of or the full-length of the instant sequence. The resulting amplification products can be labeled directly during amplification reactions or labeled after amplification reactions, and used as probes to isolate full-length DNA fragments under conditions of appropriate stringency.
[0146] Alternatively a nucleic acid sequence of the invention may be employed as a hybridization reagent for the identification of homologs. The basic components of a nucleic acid hybridization test include a probe, a sample suspected of containing the gene or gene fragment of interest, and a specific hybridization method. Probes are typically single stranded nucleic acid sequences which are complementary to the nucleic acid sequences to be detected. Probes are hybridizable to the nucleic acid sequence to be detected. The probe length can vary from 5 bases to tens of thousands of bases, and will depend upon the specific test to be done. Typically a probe length of about 15 bases to about 30 bases is suitable. Only part of the probe molecule need be complementary to the nucleic acid sequence to be detected. In addition, the complementarity between the probe and the target sequence need not be perfect. base. Hybridization methods are well defined and know in the art.
[0147] An important component of these engineered operons is the promoter sequence. The promoter must be chosen based on its compatibility with the transcriptional machinery of the host organism, as well as its ability to tune the desired level of gene expression (e.g., high or low). For example, one may introduce the strong pmxaF or pmmoX promoters from a methanotroph to generate high expression levels in a methanotrophic or non-methanotroph host. Alternatively, one can introduce a promoter from the Anderson promoter collection, which is a library of constitutive sigma70 bacterial promoters (http://partsregistry.org/Promoters/Catalog/Anderson; Registry of Standard Biological Parts), such as J23100 (strong) or J23115 (weak), to modulate expression of different ORFs or combinations of ORFs. Inducible promoters, whose activity is controlled by the addition of exogenous small molecule activators, such as IPTG, arabinose or salicylate, can also be used to provide temporal control of gene expression. However, regardless of the choice of promoter, its effect on host expression must be empirically tested in vivo to be certain of its effectiveness for achieving the desired level of expression.
[0148] These different combinatorial permutations of the cassette can be synthesized, cloned and expressed in the target host organism (via chemical transformation, electroporation, or conjugation of the DNA) so that the production of a multi-carbon product can be compared. The best candidate or candidates can then be further engineered to provide additional improvements in product yield by repeating the design-build-test cycle.
[0149] In one embodiment, the host microorganism for expressing the plasmid is a methanotroph, and plasmid vector(s) containing the metabolic pathway expression cassettes are readily mobilized into these organisms via conjugation. Various methods for bacterial conjugation are known in the art, and one of the most widely used methods takes advantage of a strain of E. coli S17-1, which has an RP4 plasmid (with the RK2 tra genes for transfer of genetic material) inserted into the chromosome for mobilizing oriT (RP4)-carrying plasmids (Simon et al. 1983; Simon, 1984).
[0150] The transfer of plasmid containing RP4-mob from E. coli to methanotrophs, as further described in the Examples section, was based on the conjugation methods described previously (Martin & Murrell, 1995; Ali, 2006). A 10 ml overnight E. coli S17-1 pir culture, containing RP4-mob plasmid, was collected on a 0.2 m pore-size nitrocellulose filter (Millipore). The E. coli donor strain was washed twice with 50 ml NMS. A 50 ml methanotroph culture grown to mid exponential phase (A.sub.540 of 0.2-0.5) was also collected on the same filter and washed again with 50 mL NMS medium. The filter was placed on an NMS agar plate containing 0.02% (w/v) proteose peptone and incubated for 24 hours at 30 C. with methane except for M. capsulatus, which was incubated at 37 C. for 24 hours.
[0151] Following incubation, the cells were washed with 10 ml NMS and collected by centrifugation (7,000g for 10 min) before re-suspending the cells in 1 ml NMS. Aliquots (50-100 l) of the cells were spread onto NMS plates containing selective antibiotics and incubated at the appropriate temperature. Colonies typically formed on the plates after 8-12 days. (Note: the E. coli S17-1 pir strain has chromosomally integrated conjugal transfer functions, thus allowing transfer of plasmid to occur by means of a bi-parental mating without a helper plasmid). Transconjugants can also be purified by serial cultivation in liquid medium containing the appropriate antibiotics for selection, followed by plating onto selective NMS agar plates to obtain single colonies.
[0152] In an alternative method for expressing metabolic pathway genes in a microbial host, the biosynthetic pathway genes are inserted directly into the chromosome. Methods for chromosomal modification include both non-targeted and targeted deletions and insertions. For example, non-targeted insertions can be achieved by using transposon mutagenesis to make insertion mutants or gene knockouts in vitro using the EZ-Tn5 <KAN-2> Insertion Kit (Epicentre). Briefly, the procedure is as follows, according to the manufacturer: Preparation: prepare 0.2 g of recombinant DNA for the EZ-Tn5<KAN-2> insertion reaction. Day 1: perform the 2-hour in vitro EZ-Tn5<KAN-2> insertion reaction; transform competent recA-E. coli with 1 l of the reaction mix and select for kanamycin-resistant transposon insertion clones on kanamycin plates overnight. Day 2: prepare DNA from kanamycin-resistant colonies, (and optionally map the EZ-Tn5<KAN-2> Transposon insertion sites and optionally (DNA) sequence chosen clones bi-directionally using the unlabeled forward and reverse transposon-specific primers supplied in the kit.
[0153] For targeted modifications, various methods have been developed based on RecA-dependent homologous recombination (Hamilton et al., 1989; Link et al., 1997; Posfai et al., 1999). However, using antibiotic resistance markers for deletion/insertion is limited by the number of different antibiotics that can be used in a given target organism. For this reason, markerless insertion methods have been developed. For example, Yu et al. (2008) describe a deletion procedure in which expression of the A-Red recombinase genes (gam, bet and exo) and the I-SceI endonuclease gene are controlled by tightly regulated promoters ParaB and PrhaB. Arabinose and rhamnose added to cultures to induce ParaB and PrhaB are used and depleted by the bacteria. Thus, by changing the carbon source in the medium from arabinose to rhamnose in bacteria that contain the pREDI plasmid, one can replace a targeted genomic region with a markerless deletion cassette and subsequently delete the selection markers that were introduced.
[0154] Sun et al. (2008) also describe methods for sequence-specific insertion or deletion of genes within a bacterial genome. This method permits multiple markerless insertions and scarless deletions in the targeted genome. In the Sun et al. method, a target gene can be deleted in two steps. In the first step, a linear DNA fragment is created that carries the cat (chloramphenicol resistance) gene and sacB (a levansucrase gene that confers sensitivity to sucrose). The fragment is flanked by long (500 bp) regions of DNA that are homologous to the regions that flank the targeted deletion site. The DNA fragment is electroporated into the host cell, which already contains plasmid pKD46, a vector containing the genes for Red recombination. Homologous recombination then directs the replacement of the targeted gene. Medium containing chloramphenicol is used to select for cells that contain the desired insertion or deletion. In the second step, a second DNA fragment that contains the desired deletion or insertion is electroporated into host cells that contain the pKD46 plasmid. By plating the resulting cells on medium containing sucrose, one can select for transformants in which the cat-sacB cassette has been replaced. These candidates are also screened for sensitivity to chloramphenicol, and the deletion can be confirmed by PCR and sequencing. By repeating the process, multiple deletions and/or insertions can be achieved. The pKD46 plasmid can then be removed by culturing the cells at 37 C. Thus, by using various genes encoding the isobutanol, butanol, fatty alcohol and fatty acid ester biosynthetic pathways, these pathways can be inserted into the genome of a methanotroph (or non-methanotroph), and unwanted genes (e.g., genes that encode for enzymes that produce competing products) can be removed.
[0155] U.S. Patent Publication No. 2006/0057726 describes using sacB gene and the pGP704 suicide vector to engineer markerless insertions into single carbon (C1) metabolizing bacteria. Yomantas et al. (2010) describes methods for markerless substitutions in the genome of the methylotrophic bacterium Methylophilus methylotrophus.
[0156] Several methanotroph strains were evaluated according to the present invention as potential hosts for pathway engineering. Of the well characterized methanotroph strains, Methylosinus trichosporium OB3b (NCIMB 11131) and Methylococcus capsulatus str. Bath (NCIMB 11853) were examined for their case of transformability (via conjugation), growth rate, and suitability for industrial fermentation. Both strains can be cultivated in liquid or agar containing Nitrate Mineral Salts (NMS) medium (Whittenbury et al., 1970; Bowman, 2000). Although both strains were found to transform with approximately equal efficiency, Methylococcus capsulatus (Bath) has the advantage of growing about twice as fast as M. trichosporium (ca. 24-30 to reach saturation in shake flask growth). In addition, the ability of M. capsulatus (Bath) to grow more readily at 45 C. is an advantage in industrial cultivation, since this relatively high temperature will impede the growth of other potentially contaminating microorganisms. Furthermore, the complete genome sequence of M. capsulatus (Bath) has been published (Ward et al., 2004), and as such, manipulation of its genome via genetic engineering is readily available to one of skill in the art. Thus, in certain embodiments, M. capsulatus (Bath) is used as a model organism for further development of genetically modified host microorganisms.
[0157] Following conjugation, positive methanotroph trans-conjugants were purified on NMS agar containing the appropriate antibiotic selection (e.g., 15 g/ml kanamycin for selecting the plasmid and counter-selecting the untransformed methanotroph host cells, and 10 g/ml for counter-selecting the E. coli donor cells). Alternatively, transconjugants can be purified by serial cultivation in liquid medium containing the appropriate antibiotics for selection, followed by plating onto selective NMS agar plates to obtain single colonies. Colonies were used to inoculate small (5-10 ml) starter cultures in liquid NMS medium containing, for example, 15 g/ml kanamycin in 125-ml flasks. The flasks were stoppered with tight-fitting Suba Seals to create a closed atmosphere inside the flasks. A volume of gas corresponding to 20% of the total volume of the flask and composed of 95% methane and 5% carbon dioxide was injected via a sterile syringe and 23-gauge needle into each flask. Flasks were shaken at 200 rpm and 45 C. When these cultures achieved an optical density of A.sub.540>0.5 (after about 24 hours), a 1:100 dilution of these cells was used to inoculate 125 ml (or larger volume) cultures, and the same growth protocol was followed. Growth in shake flasks is most robust when the liquid volume is maintained at about 5-10% of the nominal volume of the flask so that good aeration of the liquid is achieved. These flasks were then used for the subsequent assays of product formation. In certain examples related to 2-KIV feeding experiments, only the ketoacid intermediate was added along with the methane and CO.sub.2 at the zero time point.
[0158] After approximately 72 hours of growth, the cultures were harvested for analysis by gas chromatography. The sealed flasks were first chilled for at least 1 hour on ice, to concentrate any volatile organic compounds from the vapor phase into the liquid phase. After opening the flasks, an aliquot of the culture was diluted 1:2 with ethyl acetate in a clean 50 ml tube to extract and concentrate the isobutanol, butanol, fatty alcohols or fatty acid esters. After vortexing or shaking (and centrifugation to separate the phases), a small volume of the organic layer (approximately 1 ml) was filtered through a 0.2 m PTFE filter, and 1 l of the purified extract was then injected into an Agilent 7890A GC equipped with a Leap Technologies (Carrboro, NC) CombiPAL autosampler for analysis. Appropriate purified standards were included to generate a standard curve and determine the concentration of the targeted product. Each measurement included a positive control and a negative control (e.g., a wild-type sample or other appropriate background control) with each sample set. Additional details of the methods used for the specific products are given in the Examples section. Strains with the highest levels of production were designated for further scale-up in 1-10 liter fermentors.
[0159] During the analysis of the engineered host strains, unexpectedly high levels of isobutanol and butanol consumption (up to 30 mM after 72 hours of growth) was observed even in wild-type cultures of M. capsulatus (Bath), and therefore it was important to find mutant strains that can produce these products at a rate that is greater than their inherent rate of consumption. In certain embodiments of the invention, the competing alcohol dehydrogenase and alcohol oxidase activities are identified, and reduced or eliminated by gene knockouts, as described above.
[0160] For initial fermentation scale-up in the 1-10 liter range, methods similar to those described in Theisen et al. (2005) and U.S. Pat. No. 4,594,324 can be used, with specific modifications for M. capsulatus (Bath). A fermentation system such as the Sartorius-Stedim Biostat A plus system (Goettingen, Germany) can be used, or other equivalent fermentation systems and methods for methanotroph fermentation (e.g., see Jiang et al., 2010). An Applikon ADI 1030 Bio Controller and ADI 1035 BioConsole (Applikon Biotechnology Inc., Foster City, CA) can also be used for the 10 liter vessel.
[0161] The starting inoculum is created by inoculating a large colony of M. capsulatus (Bath) containing the desired plasmid from a plate culture into 10 ml of sterile NMS medium containing kanamycin, as described above. After 24 to 48 hours, when the optical density (A.sub.540) of the culture is greater than 0.5, five starter flasks of NMS medium are inoculated at 1:100 dilution. The liquid volumes in these starter inocula can range in size from 20 ml each for a 1 liter fermentor to 200 ml each for a 10 liter fermentor (i.e., about a 10% inoculum).
[0162] After autoclaving the NMS medium in the fermentor vessel, the phosphate salts portion of the NMS medium and the kanamycin (both sterilized) are added to the vessel. The same inlet can be used to inject the starter cultures. Air is supplied as oil-free compressor air, and the methane carbon source is supplied from a pre-mixed tank (Airgas) containing 95% methane and 5% CO.sub.2. The air and methane are mixed to 15-20% methane using equipment that is rated intrinsically safe or explosion proof to eliminate the possibility of sparking or static electricity, which could lead to an explosion. The gas flow rate depends on the fermentor size and culture density, but a value of 0.75 liters per minute for 10 liters is typical. The gas mixture is fed into the fermentor, and the entire culture is mixed with an impeller rotating at approximately 200 rpm for agitation, the rate of which may be increased during growth. For maintenance of the culture pH at 6.8, 0.1 M HCl or 1 M NaOH is added as needed. The temperature is maintained at 45 C. by a thermostatic jacket. The effluent gas is fed through a water-jacketed condenser to reduce liquid loss at 45 C., and vented to a fume hood.
[0163] The fermentation is monitored (via pH and dissolved oxygen probes) and controlled using Sartorius BioPAT MFCS bioprocess control software (Sartorius Corp, Bohemia, NY). A dissolved oxygen concentration below 1% saturation with air (typically 0.2-0.3%) is desirable to avoid wasting methane. Periodically, small samples of the fermentation broth are removed by sterile transfer and used to measure the optical density of the culture. These samples can also be used to monitor product formation using the methods described above and in the Examples section. Purity of the culture can also be checked by plating a small sample onto R2A agar, which allows most organisms other than methanotrophs to grow. Cultures achieve an optical density (A.sub.540) of greater than 9 after about 48 hours. For M. capsulatus (Bath), 1 ml of culture with A.sub.540 equal to 1 corresponds to about 0.23-0.25 mg of dry weight of biomass. When the maximum cell density or product concentration is achieved, the culture can be harvested and analyzed.
[0164] For large-scale commercial fermentation, a system based on the fermentor design employed by Norferm (Norefem, AS; Stavanger, Norway) for production of single-cell protein can be used (Bothe et al., 2002; EP 1419234; U.S. Publication No. 2009/0263877). The largest system has a total volume of 300 m.sup.3 (300,000 liters) and an annual production capacity of 10,000 tons of biomass (van Laere et al., 2005). Publications such as EP 1419234, U.S. Publication No. 2009/0263877 and Villadsen (2012), and references therein, describe a loop reactor and bioprocess methods for culturing methanotrophs at the commercial scale. The advantage of this design is that nutrient gases such as methane and oxygen are supplied to the system in such a way that exposure of the cells to nutrient-depleted culture medium or to unduly high concentrations of nutrient gases is minimized.
[0165] However, when using wet natural gas as a nutrient feedstock, the problem of acetate and propionate toxicity (resulting from the oxidation of ethane and propane, respectively) may need to be addressed (Bothe et al., 2002; Eiteman & Altman, 2006). A genetic approach is to eliminate (knock-out) or knock-down the ethanol and propanol dehydrogenases and acetaldehyde/propionaldehyde dehydrogenases that convert the ethanol and propanol to the corresponding acids. Another approach is to introduce the genes for acetate assimilation from an organism that can use it as a carbon source, such as E. coli (Wolfe, 2005). For example, AMP-ACS (acetate: CoA ligase [AMP forming]; EC 6.2.1.1) catalyzes the conversion of acetate and ATP to an enzyme-bound acetyladenylate (acetyl-AMP) and pyrophosphate. In a subsequent step, it reacts the acetyl-AMP with CoASH (CoenzymeA-SH) to acetyl-CoA and free AMP. Similarly, AMP-ACS can activate and assimilate propionate (Wolfe, 2005). In this way, the two potentially harmful organic acids are converted into the useful intermediate, acetyl-CoA. These genes can be cloned and expressed in a methanotroph host by the methods described above.
[0166] Another aspect of the commercial production of multicarbon compounds from methane using the present invention involves recovering and purifying the desired product from the fermentation broth. The method to be used depends on the physico-chemical properties of the product and the nature and composition of the fermentation medium and cells. For example, U.S. Pat. No. 8,101,808 describes methods for recovering C3-C6 alcohols from fermentation broth using continuous flash evaporation and phase separation processing. Thus, the biologically produced multi-carbon compounds of the invention may be isolated from the fermentation medium using methods known in the art for Acetone-butanol-ethanol (ABE) fermentations For example, solids may be removed from the fermentation medium by centrifugation, filtration, decantation, wherein the multi-carbon compounds of the invention may be isolated from the fermentation medium using methods such as distillation, azeotropic distillation, liquid-liquid extraction, adsorption, gas stripping, membrane evaporation, or pervaporation.
[0167] In certain embodiments the invention, the fermentation process produces greater than about 7% (v/v) concentration of the desired multi-carbon product in the fermentation broth, and the product is separated from the rest of the medium using membrane separation technology to achieve about a 12% or greater concentration of the product, at which point relatively small molecules (such as isobutanol) can be further purified by phase separation in an integrated system (Hickey & Slater, 1990; Neel, 1995; Hgg, 1998; Liu et al., 2011). Continuous recovery of the product from the fermentation medium has the advantage of possibly reducing the toxicity effects of the multi-carbon products.
[0168] For longer-chain alcohols, such as fatty alcohols, U.S. Pat. No. 8,268,599 describes methods for separating these components from the aqueous phase of the fermentation by bi-phasic separation, whereby the immiscibility of the product compounds with the fermentation broth allows the organic phase to be collected and removed. This separation can also reduce the toxic effects of the product on the host microbial cells.
[0169] U.S. Publication No. 2007/0251141 describes methods for recovering fatty acid methyl esters (FAMEs) from a liquid suspension by adding urea and creating a phase separation whereby the saturated and unsaturated FAMEs can be recovered separately. Membrane separation methods can also be applied to purifying fatty acid ester products such as biodiesel (Saleh, 2011).
[0170] In certain embodiments, a methane substrate of the invention is provided or obtained from a natural gas source, wherein the natural gas is wet natural gas or dry natural gas. Natural gas is referred to as dry natural gas when it is almost pure methane, having had most of the other commonly associated hydrocarbons removed. When other hydrocarbons are present, the natural gas is referred to as wet. Wet natural gas typically comprises about 70-90% methane, about 0-20% ethane, propane and butane (combined total), about 0-8% CO.sub.2, about 0-5% N2, about 0-5% H.sub.2S and trace amounts of oxygen, helium, argon, neon and xenon. In certain other embodiments, a methane substrate of the invention is provided or obtained from methane emissions, or methane off-gases, which are generated by a variety of natural and human-influenced processes, including anaerobic decomposition in solid waste landfills, enteric fermentation in ruminant animals, organic solids decomposition in digesters and wastewater treatment operations, and methane leakage in fossil fuel recovery, transport, and processing systems.
[0171] Table 1 below, provides exemplary polynucleotide and polypeptide sequences for implementing various embodiments of the present invention. These sequences are not meant to limit or exclude the use of other polynucleotide sequences encoding polypeptides or enzymes useful for producing multi-carbon compounds according to the present invention. For example, one of skill in the art can search gene sequence databases (or genome databases) and/or protein sequence databases (e.g., via BLAST or other sequence search algorithms) to identify homologous polynucleotides encoding one or more enzyme activities based on the reference sequences set forth in Table 1. Alternatively, a homologous polynucleotide may be isolated directly by using all or a portion of a nucleic acid sequence set forth in Table 1 (or a primer sequence set forth below in Table 2) as DNA hybridization probes to screen libraries from any desired microorgansim and/or PCR amplify a desired polynucleotide sequence using methodology well known to those skilled in the art.
TABLE-US-00001 TABLE 1 Exemplary Nucleic Acid and Polypeptide Sequences Described in the Invention Pathway or Nucleic acid Polypeptide Reaction Gene Name SEQ Enzyme Name SEQ ID Organism isobutanol MCA1837 SEQ ID NO: 1 ALS SEQ ID NO: 2 M. capsulatus, Bath isobutanol MCA2272 SEQ ID NO: 3 KARI SEQ ID NO: 4 M. capsulatus, Bath isobutanol MCA2082 SEQ ID NO: 5 DHAD SEQ ID NO: 6 M. capsulatus, Bath isobutanol MCA0996 SEQ ID NO: 7 KDC SEQ ID NO: 8 M. capsulatus, Bath isobutanol YMR318C SEQ ID NO: 9 ADH SEQ ID NO: 10 S. cerevisiae isobutanol MtKDC SEQ ID NO: 82 KDC SEQ ID NO: 162 M. trichosporium isobutanol MtADH SEQ ID NO: 83 ADH SEQ ID NO: 163 M. trichosporium isobutanol McADH-2a SEQ ID NO: 84 ADH SEQ ID NO: 164 M. capsulatus, Bath isobutanol McADH-2b SEQ ID NO: 85 ADH SEQ ID NO: 165 M. capsulatus, Bath Isobutanol LlkivD SEQ ID NO: 86 KDC SEQ ID NO: 166 L. lactis Isobutanol ScPDC6 SEQ ID NO: 87 KDC SEQ ID NO: 167 S. cerevisiae Isobutanol ScARO10 SEQ ID NO: 88 KDC SEQ ID NO: 168 S. cerevisiae Isobutanol ScADH2 SEQ ID NO: 89 ADH SEQ ID NO: 169 S. cerevisiae Isobutanol ScPDC1 SEQ ID NO: 90 KDC SEQ ID NO: 170 S. cerevisiae isobutanol CaPDC SEQ ID NO: 91 KDC SEQ ID NO: 171 C. acetobutylicum CH.sub.4 to CH.sub.3OH MCA1798 SEQ ID NO: 11 pmoC subunit 1 SEQ ID NO: 12 M. capsulatus, Bath CH.sub.4 to CH.sub.3OH MCA1797 SEQ ID NO: 13 pmoA subunit 1 SEQ ID NO: 14 M. capsulatus, Bath CH.sub.4 to CH.sub.3OH MCA1796 SEQ ID NO: 15 pmoB subunit 1 SEQ ID NO: 16 M. capsulatus, Bath CH.sub.4 to CH.sub.3OH MCA2855 SEQ ID NO: 17 pmoC subunit 2 SEQ ID NO: 18 M. capsulatus, Bath CH.sub.4 to CH.sub.3OH MCA2854 SEQ ID NO: 19 pmoA subunit 2 SEQ ID NO: 20 M. capsulatus, Bath CH.sub.4 to CH.sub.3OH MCA2853 SEQ ID NO: 21 pmoB subunit 2 SEQ ID NO: 22 M. capsulatus, Bath CH.sub.4 to CH.sub.3OH MCA1194 SEQ ID NO: 23 mmoX SEQ ID NO: 24 M. capsulatus, Bath CH.sub.4 to CH.sub.3OH MCA1195 SEQ ID NO: 25 mmoY SEQ ID NO: 26 M. capsulatus, Bath CH.sub.4 to CH.sub.3OH MCA1196 SEQ ID NO: 27 mmoB SEQ ID NO: 28 M. capsulatus, Bath CH.sub.4 to CH.sub.3OH MCA1198 SEQ ID NO: 29 mmoZ SEQ ID NO: 30 M. capsulatus, Bath CH.sub.4 to CH.sub.3OH MCA1199 SEQ ID NO: 31 mmoD SEQ ID NO: 32 M. capsulatus, Bath CH.sub.4 to CH.sub.3OH MCA1200 SEQ ID NO: 33 mmoC SEQ ID NO: 34 M. capsulatus, Bath CH.sub.3OH to H.sub.2CO MCA0779 SEQ ID NO: 35 mxaF SEQ ID NO: 36 M. capsulatus, Bath CH.sub.3OH to H.sub.2CO MCA0780 SEQ ID NO: 37 mxaJ SEQ ID NO: 38 M. capsulatus, Bath CH.sub.3OH to H.sub.2CO MCA0781 SEQ ID NO: 39 mxaG SEQ ID NO: 40 M. capsulatus, Bath CH.sub.3OH to H.sub.2CO MCA0782 SEQ ID NO: 41 mxaI SEQ ID NO: 42 M. capsulatus, Bath CH.sub.3OH to H.sub.2CO MCA0783 SEQ ID NO: 43 mxaR SEQ ID NO: 44 M. capsulatus, Bath CH.sub.3OH to H.sub.2CO MCA0785 SEQ ID NO: 45 mxaA SEQ ID NO: 46 M. capsulatus, Bath CH.sub.3OH to H.sub.2CO MCA0786 SEQ ID NO: 47 mxaC SEQ ID NO: 48 M. capsulatus, Bath CH.sub.3OH to H.sub.2CO MCA0787 SEQ ID NO: 49 mxaK SEQ ID NO: 50 M. capsulatus, Bath CH.sub.3OH to H.sub.2CO MCA0788 SEQ ID NO: 51 mxaL SEQ ID NO: 52 M. capsulatus, Bath CH.sub.3OH to H.sub.2CO MCA0789 SEQ ID NO: 53 mxaD SEQ ID NO: 54 M. capsulatus, Bath 1-butanol MCA0354 SEQ ID NO: 55 Threonine- SEQ ID NO: 56 M. capsulatus, Bath ammonia-lyase 1-butanol MCA2275 SEQ ID NO: 57 2-ethylmalate SEQ ID NO: 58 M. capsulatus, Bath synthase 1-butanol MCA2065 SEQ ID NO: 59 Isopropyl malate SEQ ID NO: 60 M. capsulatus, Bath dehydratase, large subunit 1-butanol MCA2064 SEQ ID NO: 61 Isopropyl malate SEQ ID NO: 62 M. capsulatus, Bath dehydratase, small subunit 1-butanol VIMSS17191 SEQ ID NO: 160 tdcB SEQ ID NO: 161 E. coli 1-butanol MCA0996 SEQ ID NO: 7 KDC SEQ ID NO: 8 M. capsulatus, Bath 1-butanol YMR318C SEQ ID NO: 9 ADH SEQ ID NO: 10 S. cerevisiae 1-butanol MtKDC SEQ ID NO: 82 KDC SEQ ID NO: 162 M. trichosporium 1-butanol MtADH SEQ ID NO: 83 ADH SEQ ID NO: 163 M. trichosporium 1-butanol McADH-2a SEQ ID NO: 84 ADH SEQ ID NO: 164 M. capsulatus, Bath 1-butanol McADH-2b SEQ ID NO: 85 ADH SEQ ID NO: 165 M. capsulatus, Bath 1-butanol LlkivD SEQ ID NO: 86 KDC SEQ ID NO: 166 L. lactis 1-butanol ScPDC6 SEQ ID NO: 87 KDC SEQ ID NO: 167 S. cerevisiae 1-butanol ScARO10 SEQ ID NO: 88 KDC SEQ ID NO: 168 S. cerevisiae 1-butanol ScADH2 SEQ ID NO: 89 ADH SEQ ID NO: 169 S. cerevisiae 1-butanol ScPDC1 SEQ ID NO: 90 KDC SEQ ID NO: 170 S. cerevisiae 1-butanol CaPDC SEQ ID NO: 91 KDC SEQ ID NO: 171 C. acetobutylicum Fatty alcohol FAR SEQ ID NO: 65 FAR SEQ ID NO: 66 M. algicola Fatty acid ester Ab-wax-dgaT SEQ ID NO: 67 wax-dgaT SEQ ID NO: 68 A. baylyi Fatty acid ester Psyc_0223 SEQ ID NO: 69 PaWES SEQ ID NO: 70 P. arcticus Fatty acid ester ROP_02100 SEQ ID NO: 71 RoWES1 SEQ ID NO: 72 R. opacus Fatty acid ester ROP_13050 SEQ ID NO: 73 RoWES2 SEQ ID NO: 74 R. opacus Fatty acid ester ROP_54550 SEQ ID NO: 75 RoWES3 SEQ ID NO: 76 R. opacus Fatty acid ester ROP_26950 SEQ ID NO: 77 RoWES4 SEQ ID NO: 78 R. opacus 2,3-butanediol YAL060W SEQ ID NO: 156 Bdh1 SEQ ID NO: 157 S. cerevisiae RuMP MCA3049 SEQ ID NO: 160 HPS SEQ ID NO: 161 M. capsulatus, Bath RuMP MCA3050 SEQ ID NO: 162 HPS/PHI SEQ ID NO: 163 M. capsulatus, Bath
TABLE-US-00002 TABLE 2 Plasmid, Primer, Promoter and Gene Fragment Sequences Described in the Invention Name Nucleic acid SEQ ID pCM132 SEQ ID NO: 79 pJSvec SEQ ID NO: 80 pMZT3 SEQ ID NO: 81 JPS00082 SEQ ID NO: 92 JPS00031 SEQ ID NO: 93 JPS00032 SEQ ID NO: 94 GMV257 SEQ ID NO: 95 JPS00118 SEQ ID NO: 96 JPS00119 SEQ ID NO: 97 ESG00087 SEQ ID NO: 98 GMV251 SEQ ID NO: 99 rnpB SEQ ID NO: 100 JPS00161 SEQ ID NO: 101 JPS00162 SEQ ID NO: 102 JPS00163 SEQ ID NO: 103 JPS00164 SEQ ID NO: 104 JPS00172 SEQ ID NO: 105 JPS00173 SEQ ID NO: 106 JPS00174 SEQ ID NO: 107 JPS00176 SEQ ID NO: 108 JPS00177 SEQ ID NO: 109 JPS00157 SEQ ID NO: 110 JPS00178 SEQ ID NO: 111 Me-AM1 PmxaF SEQ ID NO: 112 JPS00169 SEQ ID NO: 113 GMV00251 SEQ ID NO: 114 JPS00170 SEQ ID NO: 115 JPS00171 SEQ ID NO: 116 JPS00153 SEQ ID NO: 117 JPS00151 SEQ ID NO: 118 JPS00154 SEQ ID NO: 119 JPS00183 SEQ ID NO: 120 JPS00185 SEQ ID NO: 121 J23100 SEQ ID NO: 122 J23100 hybrid SEQ ID NO: 123 J23115 SEQ ID NO: 124 GMV00233 SEQ ID NO: 125 GMV00235 SEQ ID NO: 126 GMV00433 SEQ ID NO: 127 GMV00434 SEQ ID NO: 128 GMV00435 SEQ ID NO: 129 GMV00436 SEQ ID NO: 130 GMV00437 SEQ ID NO: 131 GMV00438 SEQ ID NO: 132 GMV00439 SEQ ID NO: 133 GMV00440 SEQ ID NO: 134 GMV00441 SEQ ID NO: 135 GMV00442 SEQ ID NO: 136 ESG00084 SEQ ID NO: 137 ESG00088 SEQ ID NO: 138 pMZT37 SEQ ID NO: 139 MaFAR-g1 SEQ ID NO: 140 MaFAR-g2 SEQ ID NO: 141 MaFAR-g3 SEQ ID NO: 142 MaFAR-g4 SEQ ID NO: 143 GMV410 SEQ ID NO: 144 GMV411 SEQ ID NO: 145 GMV412 SEQ ID NO: 146 GMV413 SEQ ID NO: 147 GMV414 SEQ ID NO: 148 GMV415 SEQ ID NO: 149 GMV416 SEQ ID NO: 150 GMV417 SEQ ID NO: 151 GMV418 SEQ ID NO: 152 GMV419 SEQ ID NO: 153 GMV420 SEQ ID NO: 154 GMV421 SEQ ID NO: 155 GMV422 SEQ ID NO: 158 GMV423 SEQ ID NO: 159
EXAMPLES
[0172] The present invention is further defined in the following Examples. It should be understood that these examples, while indicating certain embodiments of the invention, are given by way of illustration only. From the above discussion and these Examples, one skilled in the art can ascertain the essential characteristics of this invention, and without departing from the spirit and scope thereof, can make various changes and modifications of the invention to adapt it to various uses and conditions.
Example 1
Biosynthetic Production of Isobutanol from Methane
[0173] Initial experiments were performed to confirm and validate enzymatic activity of isobutanol pathway enzymes at the relatively high temperatures (i.e., 45 C.) requisite for growth of one preferred methanotroph host organism, Methylococcus capsulatus (Bath). Thus, in this example, the methanotroph M. capsulatus was engineered in the first series of experiments to overexpress two isobutanol pathway enzymes, ketoacid decarboxylase (KDC) and alcohol dehydrogenase (ADH), prior to introducing the full complement of five isobutanol pathway enzymes (Atsumi et al., 2010) into M. capsulatus. Following the functional validation of KDC and ADH activity in M. capsulatus (set forth below), the complete five-gene isobutanol pathway was introduced into M. capsulatus, the results of which are set forth below.
Gene Selection, Synthesis and Cloning
[0174] For the two-gene (isobutanol) pathway experiments (and for the downstream section of the five-gene isobutanol pathway set forth below), ketoacid decarboxylase (KDC) and alcohol dehydrogenase (ADH) genes were amplified by colony PCR from Methylosinus trichosporium (strain: OB3b, National Collection of Industrial, Food and Marine Bacteria (NCIMB) Accession No: 11131) and Methylococcus capsulatus (Bath). The Methylosinus trichosporium gene, MtKDC, encoding KDC is set forth in SEQ ID NO: 82, Methylosinus trichosporium gene, MtADH, encoding ADH is set forth in SEQ ID NO: 83. The Methylococcus capsulatus (Bath) gene, McKDC, encoding KDC is set forth in SEQ ID NO:7, the Methylococcus capsulatus (Bath) genes, McADH-2a and McADH-2b, encoding two ADH2 homologs, are set forth in SEQ ID NO:84 and SEQ ID NO:85, respectively.
[0175] Other KDC and ADH genes such as L1KIVD: Lactococcus lactis KDC (SEQ ID NO: 86); ScPDC6: Saccharomyces cerevisiae PDC6 (SEQ ID NO:87); ScARO10: S. cerevisiae ARO10 (SEQ ID NO:88); ScADH2: S. cerevisiae ADH2 (SEQ ID NO:89); ScPDC1: S. cerevisiae PDC1 (SEQ ID NO:90); CaPDC: Clostridium acetobutylicum PDC (SEQ ID NO:91) were codon optimized for expression in M. capsulatus and de novo synthesized by GenScript (Piscataway, NJ). Various KDC and ADH combinations were cloned with a constitutive promoter (J23115) or inducible (Ptrc) promoter into plasmid pCM132 (Accession No. AF327720; SEQ ID NO:79) with the Clontech In-Fusion kit (Mountain View, CA). A gene for the ds-Red protein was used as a control. Plasmids were transformed into E. coli S17-1 for conjugation.
Vector Inserts
[0176] Vector inserts contain the DNA fragments that are to be carried in the plasmid. The vector inserts were designed as exchangeable parts to the vector backbone described above. In one embodiment of the 2-gene pathway example, the plasmids were designed to contain two inserts made up of Methylococcus capsulatus KDC (MCA0996; SEQ ID NO:7) and Saccharomyces cerevisiae ADH6 (YMR318C; SEQ ID NO:9) genes. Both genes were amplified from genomic DNA of their respective hosts, with the primers described above in Tables 2 and below in Table 3.
TABLE-US-00003 TABLE 3 Plasmid insert modules, templates and primers Modules Plasmid PCR rxn: Vector Backbone Insert 1 Insert 2 Insert 3 pJS0025 template pJSvec MCA0996 (M. YMR318C (S. capsulatus DNA) cerevisiae DNA) primer 1 JPS0082 JPS0032 JPS00118 primer 2 JPS0031 GMV00257 JPS00119 pGMV145 template pMZT3 MCA0996 (M. YMR318C (S. capsulatus DNA) cerevisiae DNA) primer 1 JPS0082 GMV00251 JPS00118 primer 2 ESG00087 GMV00257 JPS00119 pJS034 template pGMV145 IDT gBlock synthesized rnpB DNA primer 1 JPS00161 JPS00163 primer 2 JPS00162 JPS00164 pJS041 template pJS034 MCA1837 (M. MCA2272 (M. MCA2082 pJS041n capsulatus DNA) capsulatus DNA) (M. capsulatus DNA) primer 1 JPS00162 JPS00173 JPS00176 JPS00157 primer 2 JPS00172 JPS00174 JPS00177 JPS00178 pJS048 template pJS034 IDT gBlock synthesized Me-AM1 PmxaF DNA primer 1 JPS00169 JPS00170 primer 2 GMV251 JPS00171 pJS038 template pGMV145 MCA1837 (M. capsulatus DNA) primer 1 JPS00153 JPS00151 primer 2 GMV251 JPS00154 pJS042 template pJS048 pJS038 pJS042n primer 1 JPS00162 JPS00173 primer 2 JPS00172 JPS00178 pJS050 template pJS041n pJS041n primer 1 JPS00183 JPS00174 primer 2 JPS00176 JPS00185
[0177] The modular parts (i.e., vector backbone and vector inserts) were PCR amplified (as listed in Table 3) with NEB Phusion master mix (New England Biolabs; Ipswich, MA) according to the manufacturer's instructions and in vitro assembled with the Clontech InFusion HD Cloning System kit (Clontech; Mountain View, CA) according to the manufacturer's instructions to generate circular plasmid listed below.
[0178] The in vitro assembled plasmids (2 l of the InFusion reaction) were transformed into chemically competent NEB Turbo E. coli cells, screened by colony PCR, purified, and subsequently sequence verified.
[0179] The plasmid pJSvec (SEQ ID NO:80) served as the template for the vector backbone with an inducible promoter and consisted of the pCM132 cloning vector (SEQ ID NO: 79), lacIq, and the IPTG-inducible pTrc promoter.
[0180] The plasmid pMZT3 (SEQ ID NO:81) served as the template for the vector backbone with a constitutive promoter and consisted of the pCM132 (SEQ ID NO:79) cloning vector and E. coli J23115 promoter (SEQ ID NO:124).
[0181] The plasmid pJS0025 was designed to express M. capsulatus KDC (MCA0996; SEQ ID NO:7) and S. cerevisiae ADH6 (YMR 318C; SEQ ID NO:9) from the inducible promoter.
[0182] The plasmid pGMV145 was designed to express M. capsulatus KDC (MCA0996; SEQ ID NO:7) and S. cerevisiae ADH6 (YMR 318C; SEQ ID NO:9) from the constitutive promoter.
[0183] The plasmid pJS034 introduced a second terminator sequence into pGMV145. The pGMV145 vector backbone was PCR amplified with primers JPS00161 (SEQ ID NO: 101)/JPS00162 (SEQ ID NO:102) and KOD mastermix. The insert contained DNA sequence for rnpB (SEQ ID NO:100) synthesized as a gBlock from Integrated DNA Technologies (Coralville, IA) and amplified with JPS00163 (SEQ ID NO:103)/JPS00164 (SEQ ID NO:104) primers.
Expression of the Full Five-Gene Pathway for Methane-to-Isobutanol Conversion
[0184] In order to synthesize isobutanol from methane (i.e., via pyruvate), without the need to exogenously supply a ketoacid intermediate, the pJS041 and pJS04In plasmids were designed to express all five isobutanol pathway genes: (1) M. capsulatus KDC (MCA0996; SEQ ID NO:7) and (2) S. cerevisiae ADH6 (YMR318C; SEQ ID NO:9) from the J23115 constitutive promoter (SEQ ID NO:124), and (3) M. capsulatus ilvK (MCA1837; SEQ ID NO: 1), (4) M. capsulatus ilvC (MCA2272; SEQ ID NO:3), and (5) M. capsulatus ilvD (MCA2082; SEQ ID NO:5) from the J23100 constitutive promoter (see,
TABLE-US-00004 TABLE4 Sequencecomparisonbetweenthehybrid promotersinplasmidspJS041andpJS042and thecanonicalpromotersJ23115andJ23100 J23115 TTTATAGCTAGCTCAGCC (SEQIDNO:124) CTTGGTACAATGCTAGC pJS041-hybrid TTGACGGCTAGCTCAGCC (SEQIDNO:123) CTTGGTACAATGCTAGC J23100 TTGACGGCTAGCTCAGTC (SEQIDNO:122) CTAGGTACAGTGCTAGC
[0185] The pJS048 plasmid replaced the J23100 promoter with the MxaF promoter (SEQ ID NO: 112) from Methylobacterium extorquens AM-1 in pJS034.
[0186] The pJS050 plasmid was designed to express five genes: M. capsulatus KDC (MCA0996; SEQ ID NO:7) and S. cerevisiae ADH6 (YMR318C; SEQ ID NO:9) from the J23115 constitutive promoter and M. capsulatus ilvK (MCA1837; SEQ ID NO:1), M. capsulatus ilvC (MCA2272; SEQ ID NO:3), and M. capsulatus ilvD (MCA2082; SEQ ID NO: 5) from the J23115 constitutive promoter.
Conjugations
[0187] The method for conjugal transfer of RP4-mob-containing plasmids into M. trichosporium and M. capsulatus (Bath) was based on the method described previously (Martin & Murrell, 1995; Stafford et. al., 2003). Briefly, 10 ml of a 16 hour culture of E. coli S17-1 carrying the plasmid was collected on a sterile 47 mm, 0.2 m pore-size, nitrocellulose filter (Millipore). The cells were washed with 50 ml NMS medium without antibiotic. A fresh 50 ml culture of the M. trichosporium or M. capsulatus (Bath) recipient grown to an optical density (A.sub.540) of 0.2-0.4 (mid-exponential phase of growth) was collected on the same filter as the E. coli S17-1 host cells. The cells were washed with 50 ml NMS and the filter was placed on an NMS agar plate supplemented with 0.02% (w/v) Proteose Peptone (Difco Laboratories, Detroit, MI) and incubated for 24 hours at 30 C. (for M. trichosporium) or 37 C. (M. capsulatus (Bath)) in the presence of 20-25% methane (CH.sub.4) (v/v) in air. After incubation, the cells from the conjugation plate were washed from the filter with 10 ml of NMS, pelleted by centrifugation at 7,000g, and re-suspended in 1 ml of NMS. 150 l aliquots were spread onto selective NMS plates containing 10 g/ml nalidixic acid to select against E. coli and 15 g/ml kanamycin for plasmid selection and incubated at 30 C. or 45 C. for M. trichosporium or M. capsulatus, respectively. The remaining cells were grown in NMS liquid containing 10 g/ml nalidixic acid and 15 g/ml kanamycin (Sigma, St. Louis, MO) as a secondary selection process. Cells grown in liquid selection were serially passaged three times, before spreading onto selective NMS plates for clone isolation.
M. capsulatus Growth Conditions
[0188] From a saturated starter culture, M. capsulatus (Bath) cells were diluted 1:100 into 10 ml of fresh NMS containing 15 g/ml kanamycin in a 125-ml shake flask. For ketoacid feeding experiments, cultures were treated with 1 g/L 2-ketovalerate (CAS #1821 Feb. 9) or 8 g/L 2-ketoisovalerate (CAS #3715-19-5) with or without the inducer, 0.1 mM isopropylthiogalactoside (IPTG). The flasks were closed with Suba-seals, injected with 20-25% CH.sub.4 (v/v) in air, and incubated at 45 C. for 0-120 hours.
Extraction of Alcohols from the Growth Medium
[0189] 1. Isobutanol production: The shake-flask samples were prepared for extraction by cooling them on ice for 1 hour, which ensures that the volatile organic compounds (VOC's) in the vapor phase were not lost to the atmosphere after the Suba-seal is opened.
[0190] 2. If extracting from a 9-10 ml culture, all of the culture was transferred to a 50 ml tube. For samples with high isobutanol productions (e.g., pGMV 145), 10 ml of ethyl acetate was added for extraction. For samples with low isobutanol production, only 3 ml of ethyl acetate was used. Once ethyl acetate was added to the cultures, they were subjected to either vortexing (1-2 minutes) or shaking at room temperature (for 1 hour) for efficient extraction.
[0191] 3. The tubes were then centrifuged at 4000 rpm for 20 minutes in an Eppendorf 5810 centrifuge equipped with an A-4-81 rotor.
[0192] 4. One (1) ml of the organic layer was then filtered (0.2 m PTFE membrane) and transferred to 2 ml glass Agilent gas chromatography vials for analysis.
GC-FID Analysis for Isobutanol
[0193] The extracted alcohol compounds were quantified with the Agilent 7890A gas chromatograph (GC) with flame ionization detector and PAL auto-sampler. An HP InnoWax capillary column (30 m, 0.32-mm internal diameter, 0.25-mm film thickness; Agilent Technologies, Santa Clara, CA) was used to separate the alcohols. The GC oven temperature was initially set at 35 C. for 1 minute and ramped at rate of 10 C./minute until 85 C. was reached and held for 1 minute. A second temperature ramp of 80 C./minute up to 240 C. was performed and held for 2 minutes. Hydrogen gas was the carrier gas used with 9.3 psi constant inlet pressure. The inlet and detector were maintained at 240 C. A 1 l sample was injected in split injection mode with a 25:1 split ratio.
[0194] When the two-gene KDC/ADH pathway was expressed in M. capsulatus and the isobutanol production was measured (using exogenous 2-KIV feeding), the following results were observed. A concentration of 2-KIV greater than about 4 g/L had a toxic effect on growth, wherein a 2-KIV concentration of about 2 g/L yielded the best results (
[0195] The best two-gene combination with a constitutive promoter (J23115; SEQ ID NO: 124) was M. capsulatus KDC and S. cerevisiae ADH6 (plasmid pGMV145), wherein harvesting after 48-72 hours produced the most isobutanol (
[0196] When the complete five-gene isobutanol pathway was introduced into a host strain, plasmid pJS041 yielded the highest levels of isobutanol production, with a measured titer of about 0.001 g/liter (
[0197] In certain embodiments, the production of isobutanol from methane substrate in a host strain (i.e., expressing the five-gene isobutanol pathway, e.g. via plasmid pJS041) is further optimized by genetic manipulations described above, as well as by cultivating the host strain in a fermentor culture with continuous CH.sub.4 perfusion, instead of batch addition of CH.sub.4 to the culture medium (as was done for the shake flasks experiments). In other embodiments, the production of increased isobutanol titers from methane in a host strain is further optimized via manipulations to the fermentation process (batch fed or perfusion), such as feeding additional media components as they are depleted (phosphate, nitrate, etc.) and maintaining the pH by continuously adding acid or base.
Example 2
Biosynthetic Production of 1-Butanol from Methane
[0198] A ketoacid pathway analogous to that described in Example 1, but designed to produce 1-butanol (n-butanol) is engineered in a single carbon (C1) metabolizing microbial host, such as M. capsulatus (Bath). In this example, L-threonine (which is ultimately generated from methane via phospoenolpyruvate) is first de-aminated to 2-ketobutyrate (2-oxobutanoate) by the action of threonine dehydratase (also referred to in the art as threonine ammonia-lyase (EC 4.3.1.19) encoded by the genes ilvA or tdcB) (Shen & Liao, 2008). The tdcB gene product has the biotechnological advantage that the enzyme is a catabolic enzyme, and is not feedback inhibited by L-valine or L-isoleucine (Guillouet et al., 1999).
[0199] In the second reaction step, the reaction catalyzed by leuA (encoding isopropylmalate synthase/2-ethylmalate synthase (EC 2.3.3.6)) combines 2-ketobutyrate, acetyl-CoA, and H.sub.2O to create (R)-2-ethylmalate. In the third reaction step, the gene product of leuC and leuD (encoding the two subunits of isopropylmalate isomerase) converts 2-ethylmalate into 3-ethylmalate. In the fourth reaction step, the gene product of leuB (encoding the enzyme 3-isopropylmalate dehydrogenase) converts 3-ethylmalate into 2-ketovalerate). At this stage, the same two enzymes used in the previous example, KDC (acting as a 2-ketovalerate decarboxylase) and ADH2 (alcohol dehydrogenase), are used to convert 2-ketovalerate into 1-butanol.
[0200] An alternate pathway (the citramalate pathway) from phosphoenolpyruvate and pyruvate to 2-ketobutyrate has also been described for making 1-butanol (Atsumi & Liao, 2008).
[0201] As described, above, the plasmids generated in this study are based on the broad-host-range pCM132 (Accession No. AF327720, SEQ ID NO:79) cloning vector described by Marx & Lidstrom (2001). In this embodiment, the use of the Clontech (catalog no. 639647) InFusion HD Cloning System kit is one example of how to construct plasmids, but is not meant to limit or exclude other methods that are known in the art.
Vector Backbones
[0202] Vector backbones contain the components of the plasmid that will remain constant. The broad-host range pCM132 vector was modified to produce vector backbones for the plasmids in this study. The pCM132 vector consisted of the following components: trrnB terminator, kanamycin resistance gene, trfA, IncP oriT, IncP oriV, colE1 ori, and lacZ. This parental vector was modified to replace lacZ with a vector insert that contains promoter sequence to produce plasmids pMZT3 (SEQ ID NO:81) and pMZT37 (SEQ ID NO: 139).
Vector Inserts
[0203] Vector inserts contain DNA to be added to a vector backbone. The inserts were designed as exchangeable (modular) parts to the vector and in this example consist of Methylococcus capsulatus KDC (MCA0996; SEQ ID NO:7), leuA (MCA2275; SEQ ID NO: 57), leuCDB (MCA2063; SEQ ID NO:63, MCA2064; SEQ ID NO:61 and MCA2065; SEQ ID NO:59), Saccharomyces cerevisiae ADH6 (YMR318C; SEQ ID NO:9), and M. capsulatus ilvA (MCA0354; SEQ ID NO:55) or E. coli tdcB (SEQ ID NO:160) genes. The genes were amplified from genomic DNA of their respective hosts with the primers described in Table 5.
[0204] The modular parts (vector backbone and vector insert) were PCR amplified as listed in Table 4 with NEB Phusion master mix according to the manufacturer's instructions and in vitro assembled with the Clontech InFusion HD Cloning System kit according to the manufacturer's instructions to generate circular plasmid. The in vitro assembled plasmids (2 l of the InFusion reaction) were transformed into chemically competent NEB Turbo E. coli cells, screened for by colony PCR, purified, and subsequently sequence verified.
[0205] The pGMV145 plasmid was designed to express M. capsulatus KDC (MCA0996; SEQ ID NO:7) and S. cerevisiae ADH6 (YMR318C; SEQ ID NO:9) from the constitutive promoter.
[0206] The pJS034 plasmid introduced a second terminator sequence into pGMV145. The pGMV145 vector backbone was PCR amplified with primers JPS00161 (SEQ ID NO: 101)/JPS00162 (SEQ ID NO:102) and KOD mastermix. The insert was rnpB DNA synthesized as a gBlock from IDT and amplified with JPS00163 (SEQ ID NO: 103)/JPS00164 (SEQ ID NO:104) primers.
[0207] The pGMV165 plasmid was designed to express 3 genes: M. capsulatus ilvA (MCA0354; SEQ ID NO:55), M. capsulatus KDC (MCA0996; SEQ ID NO:7) and S. cerevisiae ADH6 (YMR318C; SEQ ID NO:9) from the J23115 (SEQ ID NO:124) constitutive promoter.
[0208] The pGMV166 plasmid was designed to express 3 genes: E. coli tdcB (SEQ ID NO: 160), M. capsulatus KDC (MCA0996; SEQ ID NO:7) and S. cerevisiae ADH6 (YMR318C; SEQ ID NO:9) from the J23115 (SEQ ID NO:124) constitutive promoter.
[0209] The pGMV167 plasmid was designed to express 7 genes: M. capsulatus ilvA (MCA0354; SEQ ID NO:55), M. capsulatus KDC (MCA0996; SEQ ID NO:7) and S. cerevisiae ADH6 (YMR318C; SEQ ID NO:9) from the J23115 (SEQ ID NO:124) constitutive promoter and M. capsulatus leuCDB (MCA2063; SEQ ID NO:63, MCA2064; SEQ ID NO:61 and MCA2065; SEQ ID NO:59) and M. capsulatus leuA (MCA2275; SEQ ID NO: 57) from second J23115 (SEQ ID NO:124) constitutive promoter.
[0210] The pGMV168 plasmid was designed to express 7 genes: E. coli tdcB (SEQ ID NO: 160), M. capsulatus KDC (MCA0996; SEQ ID NO:7) and S. cerevisiae ADH6 (YMR318C; SEQ ID NO:9) from the J23115 constitutive promoter and M. capsulatus leuCDB (MCA2063; SEQ ID NO:63, MCA2064; SEQ ID NO:61 and MCA2065; SEQ ID NO: 59) and leuA (MCA2275; SEQ ID NO:57) from a second J23115 constitutive promoter.
[0211] Host strains modified with these plasmids were grown on methane as described in the examples above, harvested, extracted, and analyzed for 1-butanol production.
TABLE-US-00005 TABLE 5 Insert Modules, Templates and Primers for 1-Butanol Production Modules PCR Vector Plasmid rxn: backbone Insert 1 Insert 2 Insert 3 pGMV145 template pMZT3 MCA0996 (M. YMR318C (S. capsulatus DNA) cerevisiae DNA) primer 1 JPS0082 GMV00251 JPS00118 primer 2 ESG00087 GMV00257 JPS00119 pJS034 template pGMV145 IDT gBlock synthesized rnpB DNA primer 1 JPS00161 JPS00163 primer 2 JPS00162 JPS00164 pGMV165 template pJS034 pJS034 MCA0354 (M. capsulatus DNA) primer 1 GMV435 GMV433 GMV431 primer 2 ESG000087 GMV434 GMV432 pGMV166 template pJS034 pJS034 tdcB (E. coli DNA) primer 1 GMV435 GMV433 GMV436 primer 2 ESG000087 GMV434 GMV437 pGMV167 template pGMV165 pGMV165 MCA2063-2065 (M. MCA2275 (M. capsulatus DNA) capsulatus DNA) primer 1 JPS163 GMV235 GMV439 GMV441 primer 2 GMV233 GMV438 GMV440 GMV442 pGMV168 template pGMV166 pGMV166 MCA2063-2065 (M. MCA2275 (M. capsulatus DNA) capsulatus DNA) primer 1 JPS163 GMV235 GMV439 GMV441 primer 2 GMV233 GMV438 GMV440 GMV442
Example 3
Biosynthetic Production of Fatty Alcohols from Methane
[0212] Conversion of methane to diesel components requires engineering the native metabolism of methanotrophs. The two principal native pathways that can be engineered for increased production of diesel components are the fatty acid pathway and isoprenoid pathway. In the current example, the invention describes the use of the fatty acid pathway for synthesis of diesel (wax ester) components.
[0213] Fatty acids are an important source of energy and adenosine triphosphate (ATP) for many cellular organisms. Excess fatty acids, glucose, and other nutrients can be stored efficiently as fat. All cell membranes are built up of phospholipids, each of which contains fatty acids. Fatty acids are also used for protein modification. Fatty acid synthesis is the creation of fatty acids from acetyl-CoA and malonyl-CoA precursors through action of enzymes called fatty acid synthases. Fatty acid chain length and degree of saturation depends on the host microorganism. With regard to M. capsulatus (Bath), the primary fatty acids are C16 with saturated or mono unsaturated carbon chains.
[0214] The conversion of methane to diesel components requires the over-expression of specific heterologous (exogenous) enzymes within a methanotroph (or non-methanotroph) host microorganism, wherein the over-expression of specific heterologous (exogenous) enzymes can divert the flux from native fatty acid synthesis to compounds of interest. Key intermediates of the fatty acid pathway are the fatty acyl-ACP molecules. Thus, the over-expression of specific heterologous enzymes in a host microorganism divert the flux from acyl-ACP to diesel components such as fatty acids, fatty alcohols, fatty esters and derivatives thereof. Thus, in certain embodiments, a host microorganism has been engineered to over-express specific enzymes such as a fatty acyl ACP reductase (FAR), a fatty acyl CoA reductase (CAR) and wax ester synthases (WES) for diverting flux from native compounds to compounds of interest. Active expression of these enzymes results in the conversion of methane to diesel components via FARs, CARs and WES enzymes cloned and expressed in a host microorganism (e.g., M. capsulatus (Bath)).
[0215] A biosynthetic pathway analogous to that described in Example 1, but designed to produce fatty alcohols can be engineered in a (C1) metabolizing host microorganism, such as M. capsulatus. In this example, fatty acyl-CoA (which is ultimately generated from methane via pyruvate) is converted directly into fatty alcohols by the heterologous overexpression of a fatty-acyl-CoA reductase (FAR).
Construction of Methanotroph Plasmids for Fatty Alcohol Production
[0216] As described, above, the plasmids generated in this study are based on the broad-host-range pCM132 (Accession No. AF327720) cloning vector (SEQ ID NO:79) described by Marx & Lidstrom (2001). In this embodiment, the use of the Clontech (catalog no. 639647) InFusion HD Cloning System kit is one example of how to construct plasmids, but is not meant to limit or exclude other methods that are known in the art.
Vector Backbones
[0217] Vector backbones contain the components of the plasmid that will remain constant. The broad-host range pCM132 vector was modified to produce vector backbones for the plasmids in this study. The pCM132 vector consisted of the following components: trrnB terminator, kanamycin resistance gene, trfA, IncP oriT, IncP oriV, colE1 ori, and lacZ. This parental vector was modified to replace lacZ with a vector insert that contains promoter sequence to produce plasmids pMZT3 (SEQ ID NO:81) and pMZT37 (SEQ ID NO: 139).
Vector Inserts
[0218] Vector inserts contain DNA to be added to the vector backbone. The inserts were designed as exchangeable (modular) parts to the vector and in this embodiment consist of the following components. In this example, the plasmids were designed to contain one insert: Marinobacter algicola fatty acid reductase (MaFAR; SEQ ID NO:65), also known as a fatty acyl-CoA reductase. The MaFAR gene was codon optimized and synthesized as a series of 4 gBlocks from Integrated DNA Technologies (Coralville, IA). The synthesized DNA was designed to include pivot regions to allow proper assembly by InFusion.
Assembly of the Constructs
[0219] The modular parts (vector backbone and vector insert) were PCR amplified as listed in Table 4 with NEB Phusion master mix according to the manufacturer's instructions and in vitro assembled with the Clontech InFusion HD Cloning System kit according to the manufacturer's instructions to generate circular plasmid. The in vitro assembled plasmids (2 l of the InFusion reaction) were transformed into chemically competent NEB Turbo E. coli cells, screened for by colony PCR, purified, and subsequently sequence verified.
[0220] Plasmid pMZT3 (SEQ ID NO:81) served as the template for the vector backbone with a constitutive promoter and consisted of the pCM132 cloning vector, E. coli J23115 promoter. The vector backbone was PCR amplified from the pMZT3 template with primers ESG00084 (SEQ ID NO:137)/ESG00087 (SEQ ID NO:98).
[0221] Plasmid pMZT37 (SEQ ID NO:139) served as the template for the vector backbone with a constitutive promoter and consisted of the pCM132 cloning vector, E. coli J23100 promoter. The vector backbone was PCR amplified from the pMZT3 template with primers ESG00084 (SEQ ID NO:137)/ESG00088 (SEQ ID NO:138).
[0222] The pGMV147 plasmid was designed to express M. algicola FAR gene (SEQ ID NO: 65) from the J23115 constitutive promoter (SEQ ID NO:124). The modules of this plasmid included the PCR amplified pMZT3 vector backbone and four synthesized DNA gene fragments from IDT (MaFAR-g1; SEQ ID NO:140, MaFAR-g2; SEQ ID NO:141, MaFAR-g3; SEQ ID NO:142 and MaFAR-g4; SEQ ID NO:143).
[0223] The pGMV148 plasmid was designed to express M. algicola FAR gene (SEQ ID NO: 65) from the J23110 constitutive promoter (SEQ ID NO:122). The modules of this plasmid included the PCR amplified pMZT37 vector backbone and four synthesized DNA gene fragments from IDT (MaFAR-g1; SEQ ID NO:140, MaFAR-g2; SEQ ID NO:141, MaFAR-g3; SEQ ID NO:142 and MaFAR-g4; SEQ ID NO:143).
[0224] Gas chromatography results after various host strains were grown on methane in shake flasks, extracted, and analyzed as described above, are set forth in
TABLE-US-00006 TABLE 6 Insert Modules, Templates and Primers for Fatty Alcohol Production Modules Plasmid PCR rxn: Vector backbone Insert 1 Insert 2 Insert 3 Insert 4 pGMV147 template pMZT3 MaFAR-g1 MaFAR-g2 MaFAR-g3 MaFAR-g4 primer 1 ESG00084 primer 2 ESG00087 pGMV148 template pMZT37 MaFAR-g1 MaFAR-g2 MaFAR-g3 MaFAR-g4 primer 1 ESG00084 primer 2 ESG00088
Example 4
Biosynthetic Production of Fatty Acid Methyl Esters from Methane
Construction of Methanotroph Plasmids for Fatty Acid Ester (Wax Ester) Production
[0225] The plasmids generated in this example are based on the broad-host-range pCM132 (Accession no. AF327720, SEQ ID NO: 79) cloning vector described by Marx & Lidstrom (2001). In this embodiment, the use of the Clontech (catalogue no. 639647) InFusion HD Cloning System kit is one example of how to construct plasmids, but is not meant to limit or exclude other methods that are known in the art.
Vector Backbones
[0226] Vector backbones contain the components of the plasmid that will remain constant. The broad-host range pCM132 vector was modified to produce vector backbones for the plasmids in this study. The pCM132 vector consisted of the following components: trrnB terminator, kanamycin resistance gene, trfA, IncP oriT, IncP oriV, colE1 ori, and lacZ. This parental vector was modified to replace lacZ with a vector insert that contains promoter sequence to produce plasmids and pMZT3 and pMZT37.
Vector Inserts
[0227] Vector inserts contain DNA to be added to a vector backbone. The inserts were designed as exchangeable (modular) parts to the vector and in this embodiment consist of a wax ester synthase (WES) derived from Acinetobacter sp. ADP1 (SEQ ID NO:67), Psychrobacter arcticum 273-4 (SEQ ID NO:69) or Rhodococcus opcaus B4 (SEQ ID NO: 71, SEQ ID NO:73, SEQ ID NO:75 or SEQ ID NO:77). The WES genes were codon-optimized and synthesized by GenScript.
Assembly of the Constructs
[0228] The modular parts (vector backbone and vector insert) were PCR amplified as listed in Table 7 with NEB Phusion master mix according to the manufacturer's instructions and in vitro assembled with the Clontech InFusion HD Cloning System kit according to the manufacturer's instructions to generate circular plasmid. The in vitro assembled plasmids (2 l of the InFusion reaction) were transformed into chemically competent NEB Turbo E. coli cells, screened for by colony PCR, purified, and subsequently sequence verified.
[0229] Plasmid pMZT3 (SEQ ID NO:81) served as the template for the vector backbone with a constitutive promoter and consisted of the pCM132 cloning vector, E. coli J23115 promoter. The vector backbone was PCR amplified from the pMZT3 template with primers ESG00084 (SEQ ID NO:137)/ESG00087 (SEQ ID NO:98).
[0230] Plasmid pMZT37 (SEQ ID NO:139) served as the template for the vector backbone with a constitutive promoter and consisted of the pCM132 cloning vector, E. coli J23100 promoter. The vector backbone was PCR amplified from the pMZT3 template with primers ESG00084 (SEQ ID NO:137)/ESG00088 (SEQ ID NO:138).
[0231] The pGMV153 plasmid was designed to express Acinetobacter sp. ADP1 WES gene (wax-dgaT; SEQ ID NO:67) from the J23115 constitutive promoter. The modules of this plasmid included the PCR amplified pMZT3 vector backbone and the codon-optimized synthesized gene.
[0232] The pGMV154 plasmid was designed to express Psychrobacter arcticum 273-4 WES gene (Psyc_0223; SEQ ID NO:69) from the J23115 constitutive promoter. The modules of this plasmid included the PCR amplified pMZT3 vector backbone and the codon-optimized synthesized gene.
[0233] The pGMV155 plasmid was designed to express Rhodococcus opcaus B4 WES gene (ROP_02100; SEQ ID NO:71) from the J23115 constitutive promoter. The modules of this plasmid included the PCR amplified pMZT3 vector backbone and the codon-optimized synthesized gene.
[0234] The pGMV156 plasmid was designed to express Rhodococcus opcaus B4 WES gene (ROP_13050; SEQ ID NO:73) from the J23115 constitutive promoter. The modules of this plasmid included the PCR amplified pMZT3 vector backbone and the codon-optimized synthesized gene.
[0235] The pGMV157 plasmid was designed to express Rhodococcus opcaus B4 WS gene (ROP_26950; SEQ ID NO:77) from the J23115 constitutive promoter. The modules of this plasmid included the PCR amplified pMZT3 vector backbone and the codon-optimized synthesized gene.
[0236] The pGMV158 plasmid was designed to express Rhodococcus opcaus B4 WES gene (ROP_54550; SEQ ID NO:75) from the J23115 constitutive promoter. The modules of this plasmid included the PCR amplified pMZT3 vector backbone and the codon-optimized synthesized gene.
[0237] The pGMV159 plasmid was designed to express Acinetobacter sp. ADP1 WES gene (wax-dgaT; SEQ ID NO:67) from the J23100 constitutive promoter. The modules of this plasmid included the PCR amplified pMZT3 vector backbone and the codon-optimized synthesized gene.
[0238] The pGMV160 plasmid was designed to express Psychrobacter arcticum 273-4 WES gene (Psyc_0223; SEQ ID NO:69) from the J23100 constitutive promoter. The modules of this plasmid included the PCR amplified pMZT3 vector backbone and the codon-optimized synthesized gene.
[0239] The pGMV161 plasmid was designed to express Rhodococcus opcaus B4 WES gene (ROP_02100; SEQ ID NO:71) from the J23100 constitutive promoter. The modules of this plasmid included the PCR amplified pMZT3 vector backbone and the codon-optimized synthesized gene.
[0240] The pGMV162 plasmid was designed to express Rhodococcus opcaus B4 WES gene (ROP_13050; SEQ ID NO:73) from the J23100 constitutive promoter. The modules of this plasmid included the PCR amplified pMZT3 vector backbone and the codon-optimized synthesized gene.
[0241] The pGMV163 plasmid was designed to express Rhodococcus opcaus B4 WES gene (ROP_26950; SEQ ID NO:77) from the J23100 constitutive promoter. The modules of this plasmid included the PCR amplified pMZT3 vector backbone and the codon-optimized synthesized gene.
[0242] The pGMV164 plasmid was designed to express Rhodococcus opcaus B4 WES gene (ROP_54550; SEQ ID NO:75) from the J23100 constitutive promoter. The modules of this plasmid included the PCR amplified pMZT3 vector backbone and the codon-optimized synthesized gene.
[0243] Strains modified with these plasmids are grown on methane as described in the examples above, harvested, extracted, and analyzed for fatty acid ester production.
TABLE-US-00007 TABLE 7 Insert Modules, Templates and Primers for Fatty Acid Ester Production Modules Vector Plasmid PCR rxn: backbone Insert template pMZT3 pUC57-AbWS (WS-dgaT) pGMV153 primer 1 ESG00084 GMV410 primer 2 ESG00087 GMV416 pGMV154 template pMZT3 pUC57-PaWS (Psyc_0223) primer 1 ESG00084 GMV411 primer 2 ESG00087 GMV417 template pMZT3 pUC57-RoWS (ROP_02100) pGMV155 primer 1 ESG00084 GMV412 primer 2 ESG00087 GMV418 pGMV156 template pMZT3 pUC57-RoWS (ROP_13050) primer 1 ESG00084 GMV413 primer 2 ESG00087 GMV419 template pMZT3 pUC57-RoWS (ROP_26950) pGMV157 primer 1 ESG00084 GMV414 primer 2 ESG00087 GMV420 pGMV158 template pMZT3 pUC57-RoWS (ROP_54550) primer 1 ESG00084 GMV415 primer 2 ESG00087 GMV421 template pMZT37 pUC57-AbWS (WS-dgaT) pGMV159 primer 1 ESG00084 GMV410 primer 2 ESG00088 GMV416 pGMV160 template pMZT37 pUC57-PaWS (Psyc_0223) primer 1 ESG00084 GMV411 primer 2 ESG00088 GMV417 template pMZT37 pUC57-RoWS (ROP_02100) pGMV161 primer 1 ESG00084 GMV412 primer 2 ESG00088 GMV418 pGMV162 template pMZT37 pUC57-RoWS (ROP_13050) primer 1 ESG00084 GMV413 primer 2 ESG00088 GMV419 template pMZT37 pUC57-RoWS (ROP_26950) pGMV163 primer 1 ESG00084 GMV414 primer 2 ESG00088 GMV420 pGMV164 template pMZT37 pUC57-RoWS (ROP_54550) primer 1 ESG00084 GMV415 primer 2 ESG00088 GMV421
Example 5
Biosynthetic Production of 2,3-Butanediol from Methane
[0244] The four-carbon (C4) diol 2,3-butanediol is an important intermediate for the chemical industry. At the commercial scale, it is mostly generated from petroleum. It serves as a precursor for the production of various commodity and specialty chemicals, such as the solvent methyl ethyl ketone (MEK), gamma-butyrolactone (GBL), and 1,3-butadiene. The potential production of these downstream commercial products amounts to about 32 million tons per year, with a value of about $43 billion (Kpke et al., 2011).
[0245] Biological production of 2,3-butanediol from methane requires engineering the native (or endogenous) metabolism of methanotrophs to take advantage of their endogenous production of (R)-acetoin (
Construction of Methanotroph Plasmids for 2,3-Butanediol Production
[0246] As described, above, the plasmids generated in this study are based on the broad-host-range pCM132 (Accession no. AF327720, SEQ ID NO: 79) cloning vector described by Marx & Lidstrom (2001). In this embodiment, the use of the Clontech (catalogue 639647) InFusion HD Cloning System kit is one example of how to construct plasmids, but is not meant to limit or exclude other methods that are known in the art. Sequences for the ORF and PCR primers are presented below in Table 1.
Vector Backbones
[0247] Vector backbones contain the components of the plasmid that will remain constant. The broad-host range pCM132 vector was modified to produce vector backbones for the plasmids in this example. The pCM132 vector consists of the following components: trrnB terminator, kanamycin resistance gene, trfA, IncP oriT, IncP oriV, colE1 ori, and lacZ. This parental vector has been modified to replace lacZ with a vector insert that contains promoter sequence to produce plasmid pMZT3, which was used for this example.
Vector Inserts
[0248] Vector inserts contain DNA to be added to the vector backbone. The inserts were designed as exchangeable (modular) parts to the vector, and in this embodiment consists of the components listed in Table 1 and Table 8. In this example, the plasmids were designed to contain one insert: Saccharomyces cerevisiae (R,R)-butanediol dehydrogenase (Standard name: Bdh1p (EC 1.1.1.4); SEQ ID NO:156; Systematic gene name: YAL060W).
[0249] The BDH1 gene (SEQ ID NO:156) was codon optimized and synthesized by Integrated DNA Technologies (Coralville, IA).
Assembly of the Constructs
[0250] The modular parts (vector backbone and vector insert) were PCR amplified as listed in Table 8 with NEB Phusion master mix according to the manufacturer's instructions and in vitro assembled with the Clontech InFusion HD Cloning System kit according to the manufacturer's instructions to generate circular plasmid. The in vitro assembled plasmids (2 l of the InFusion reaction) were transformed into chemically competent NEB Turbo E. coli cells, screened for by colony PCR, purified, and subsequently sequence verified.
[0251] Plasmid pMZT3 served as the template for the vector backbone with a constitutive promoter and consisted of the pCM132 cloning vector, E. coli J23115 promoter. The vector backbone was PCR amplified from the pMZT3 template with primers ESG00084 (SEQ ID NO: 137)/ESG00087 (SEQ ID NO:98).
[0252] The pGMV111 plasmid was designed to express the S. cerevisiae BDH1 gene (SEQ ID NO: 156) from the J23115 constitutive promoter. The modules of this plasmid included the PCR amplified pMZT3 vector backbone and the ScBDH1 insert amplified from the shuttle vector pUC57-ScBDH1 template using primers GMV268 (SEQ ID NO: 158)/GMV271 (SEQ ID NO:159). The plasmid was conjugated from E. coli donor strain S17-1 into the M. capsulatus (Bath) recipient as described above Example 1. The transconjugant strain was purified by repeated rounds of antibiotic selection using kanamycin and naladixic acid to remove the parent cells, as described in Example 1 above.
[0253] Cells expressing the pGMV111 plasmid were cultivated in liquid NMS medium in sealed shake flasks in the presence of 20% methane at 45 C. as described above in Example 1, for about 72 hours with 200 rpm shaking. For UPLC analysis, proteins and other debris were separated from the 2,3-butanediol in the growth medium using 2% (wt/vol.) 5-sulfosalicylic acid and centrifugation as described in Kpke et al. (2011). Extracted samples can be analyzed using a BioRad (Hercules, CA) Fast Acid column on a Waters (Milford, MA) Acquity H-class UPLC equipped with a #2414 Refractive Index Detector. Other conditions are as follows: the mobile phase is 5 mM H.sub.2SO4, the flow rate is 0.4 ml/min, the column is maintained at 40 C, and the product is detected at 410 nm.
[0254] Methods for the processing of biologically produced 1,3-propanediol and 2,3-butanediol are further described by Xiu & Zeng, 2008.
[0255] For GC analysis, the 2,3-butanediol can be extracted from the culture medium with ethyl acetate, as described in Xiao et al., (2012). The extracted sample is analyzed on an Agilent (Santa Clara, CA) 7890A GC equipped with a Leap Technologies CombiPAL autosampler and a flame ionization detector. Either an Agilent HP-INNOWax or HP-5 MS GC column can be used to separate the components according to the method of Xiao et al. (2012). Alternatively, the samples can be analyzed on a Waters Acquity H-Class UPLC equipped with a Waters 2414 Refractive Index detector using a method similar to that of Kpke et al. (2011). A BioRad (Hercules, CA) Fast Acid Column operated at 40 C. with a flow rate of 0.4 ml/minute and a 5 mM H.sub.2SO4 mobile phase can be used to perform the separation. Samples for either GC or UPLC can be quantitated against a series of known concentrations of purified (D-()-, L-(+)-, and meso-)2,3-butanediol standards (Sigma, St. Louis, MO).
[0256] At the industrial fermentation scale, the 2,3-butanediol product can be extracted from the fermentation medium using one of the following methods: steam stripping, solvent extraction, aqueous two-phase extraction, reactive extraction, and pervaporation. These methods are described in Xiu & Zeng (2008).
TABLE-US-00008 TABLE 8 Modules Vector Plasmid PCR reaction: backbone Insert pGMV111 template pMZT3 pUC57- ScBDH1 primer 1 ESG00084 GMV268 primer 2 ESG00087 GMV271
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TABLE-US-00009 SequenceListing 1 SequenceListingInformation 1-1 FileName 35016-001C3.xml 1-2 DTDVersion V1_3 1-3 SoftwareName WIPOSequence 1-4 SoftwareVersion 2.3.0 1-5 ProductionDate 2024May24 1-6 Originalfreetextlanguage code 1-7 NonEnglishfreetext languagecode 2 GeneralInformation 2-1 Currentapplication: US IPOffice 2-2 Currentapplication: Applicationnumber 2-3 Currentapplication: Filingdate 2-4 Currentapplication: 35016-001C3 Applicantfilereference 2-5 Earliestpriority US application:IPOffice 2-6 Earliestpriority 61/782,830 application: Applicationnumber 2-7 Earliestpriority 2013Mar.14 application: Filingdate 2-8en Applicantname Coleman,WilliamJ 2-8 Applicantname:NameLatin 2-9 Inventorname 2-9 Inventorname:NameLatin 2-10en Inventiontitle BiologicalProductionofMulti- CarbonCompoundsFromMethane 2-11 SequenceTotalQuantity 175 3-1 Sequences 3-1-1 SequenceNumber[ID] 1 3-1-2 MoleculeType DNA 3-1-3 Length 1662 source1...1662 3-1-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-1-5 Residues atgcgtgaaacgatacctccccgcaccggcgccgacctgctggtcgactccctccaggcg 60 ctgggcgtcgaatacgtcttcggcgtgcccggcggcgcgatactcccgatcctgaacgtg 120 ctggccgaccgcggcccgcgcttcatcgtttgccgggacgaaaccggcgccgccttcatg 180 gcccagtcctggggccggatcaccggccggcccggcgtggtgctcaccacctccggcccc 240 ggcctcatcaacgccgtctgtggcgtcgctaccgccacagaggaccgcgacccgctggtc 300 gtcatcaccggccaggtgccgcgggccgtgcaattcaagcagagccacatgaacctggat 360 tcggtcggcctgttcgcgccgatcaccaaatggagcgtcgaggtcgaggaaccgaatact 420 gtatcggaaatcctggtcaacgccttccgcaccgcgcagacgccgtgcgccggagccgtc 480 cacgtctcggtaccgaacgacatgctcaccgcgccggtcaccgcgcaggccctggcgccg 540 gccgaacccgccgtctggggaacggccccggccgccgtcgtcgaacgcgcggcgtccctg 600 ctgaacgatgccaaagccccggccatcctgctcggattgcgggccagcacacctggagcg 660 gcggcggcggtccggcgtttcctggagcggcatccgctgccggtggcgatgaccttcgaa 720 gccgccggcaccctgtcccgcgatctggtcgatcagttcgtcggccgggtcggctacgtg 780 ctcaaccagccgggcgacgaggtgctgcgccaagccgatctggtactcacgatcggctac 840 gacccgatcgaatacgaaccttccgcctggatctcaccgcagtcgcaggcgatccacctg 900 gatgccctgcccgccgccgtcgaccgggcctaccaccctgccgccgaactggtcggcgac 960 atcgccgccaacctggccgcgctcggcagcctgctccgaatcgaggatcgagccggacgc 1020 cccgccgtcgccgcggcgcggcggcgtctgctggaggagcaagcccgcggcgcagcactg 1080 accggtatgccgatccaccccttgcgcttcattcacgaccttcgggccacgctggacgac 1140 gaggcgacggtgacctgcgacgtcggcgcccacgagatctggatggcccgctacttcttc 1200 tgctacgccccgcgtcacctgctgttcagcatgggccaccagaccatgggcgtcgccctg 1260 ccctgggccatcggcgcggccctggcccggcccggcaagaaagtggtttcggtatccggc 1320 gacggctccttcctcatgacctgcatggaactggaaaccgcggtgcgcctcaaactgccg 1380 atcgtgcacatcgtctggaaagacggcggctacaacctgatccacagcctgcagatgcgc 1440 gactatgggcgcagcttcggcgccgagttcggccccaccgacttcgtcaaactggcggag 1500 gccttcggcgcgatcgggtaccggatcgagtccgcggacgggatcgtccctgtgctgaac 1560 cgggcgctcgcggccgacgcgccggtgctgatcgaagtgcccatcgactacagcgacaac 1620 gtccacctggtcgaggcgatcgacgcctcggcgcagcactga 1662 3-2 Sequences 3-2-1 SequenceNumber[ID] 2 3-2-2 MoleculeType AA 3-2-3 Length 553 source1...553 3-2-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualiferValue 3-2-5 Residues MRETIPPRTGADLLVDSLQALGVEYVFGVPGGAILPILNVLADRGPRFIVCRDETGAAFM 60 AQSWGRITGRPGVVLTTSGPGLINAVCGVATATEDRDPLVVITGQVPRAVQFKQSHMNLD 120 SVGLFAPITKWSVEVEEPNTVSEILVNAFRTAQTPCAGAVHVSVPNDMLTAPVTAQALAP 180 AEPAVWGTAPAAVVERAASLLNDAKAPAILLGLRASTPGAAAAVRRFLERHPLPVAMTFE 240 AAGTLSRDLVDQFVGRVGYVLNQPGDEVLRQADLVLTIGYDPIEYEPSAWISPQSQAIHL 300 DALPAAVDRAYHPAAELVGDIAANLAALGSLLRIEDRAGRPAVAAARRRLLEEQARGAAL 360 TGMPIHPLRFIHDLRATLDDEATVTCDVGAHEIWMARYFFCYAPRHLLFSMGHQTMGVAL 420 PWAIGAALARPGKKVVSVSGDGSFLMTCMELETAVRLKLPIVHIVWKDGGYNLIHSLQMR 480 DYGRSFGAEFGPTDFVKLAEAFGAIGYRIESADGIVPVLNRALAADAPVLIEVPIDYSDN 540 VHLVEAIDASAQH 553 3-3 Sequences 3-3-1 SequenceNumber[ID] 3 3-3-2 MoleculeType DNA 3-3-3 Length 1017 source1...1017 3-3-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualiferValue 3-3-5 Residues atgcagatttactacgacaaagacgccgacctttccatcatccagggaaagaaggttgcc 60 atcatcggctacggctcgcagggccacgcccacgccaacaacctcaaggattccggagtg 120 caggtcgtggtggggctgcgtccgggttcggcttccgccaagaaggccgagaacgccggc 180 ctcgcggtcgcctcggtcgaggatgcggtcaaacaggcggacgtcatcatgatcctggcg 240 ccggacgagcatcaggcccgcctctacaatgaacagatcgcgccgaacatcaagcagggc 300 gccgccctcgccttcgcccacggcttcaacatccacttcgagcagatcaccccgcgcgcc 360 gacctcgacgtgatcatgatcgcgcccaagggtcccggccatctggtacgttccacctac 420 acccagggcggcggcgtgccctcgctgatcgccgtgtaccagaatgccagcgggcgcgcc 480 aaggaactcgcgctgtcctatgcttcggccaatggcggcggtcgggctggtatcatcgag 540 accaccttccgcgaagagaccgaaaccgatctgttcggcgaacaggccgtcctgtgtggc 600 ggcgccaccgcactggtgcaggcgggtttcgagacgctggtcgaagccggttatgcgccc 660 gagatggcctatttcgagtgtctgcacgaactcaagctgatcgtcgacctgatgtacgaa 720 ggcggcatcgccaacatgcgttattcgatctccaatacggcagagtacggcgacctgacc 780 cgtggtccgcgcatcgtcaccgagcagaccaagcaggaaatgaagaaaatcctgcgcgag 840 atccagaccggcgaattcgcccgtgagttcattttggaaaaccaggccggagccgccacc 900 ctgaaagcgaaacgccgtctcggccgagagcatctcatcgagagcgtgggcgccaggctg 960 cgcgacatgatgccgtggatcaaggccaaccgcattgtggacacgagcaagaactga 1017 3-4 Sequences 3-4-1 SequenceNumber[ID] 4 3-4-2 MoleculeType AA 3-4-3 Length 338 source1...338 3-4-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-4-5 Residues MQIYYDKDADLSIIQGKKVAIIGYGSQGHAHANNLKDSGVQVVVGLRPGSASAKKAENAG 60 LAVASVEDAVKQADVIMILAPDEHQARLYNEQIAPNIKQGAALAFAHGFNIHFEQITPRA 120 DLDVIMIAPKGPGHLVRSTYTQGGGVPSLIAVYQNASGRAKELALSYASANGGGRAGIIE 180 TTFREETETDLFGEQAVLCGGATALVQAGFETLVEAGYAPEMAYFECLHELKLIVDLMYE 240 GGIANMRYSISNTAEYGDLTRGPRIVTEQTKQEMKKILREIQTGEFAREFILENQAGAAT 300 LKAKRRLGREHLIESVGARLRDMMPWIKANRIVDTSKN 338 3-5 Sequences 3-5-1 SequenceNumber[ID] 5 3-5-2 MoleculeType DNA 3-5-3 Length 1689 source1...1689 3-5-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-5-5 Residues atgaccgacaagcacccccgtccccattcgtcccaggtcgtcgacggcatggagcgcgcc 60 ccgagccgcgcgatgctgcacgccgtcggcttcgccgatgccgacttcgccaaaccgcag 120 atcggcatcgcttccacctgggcgatggtgacgccgtgcaacatgcacatcaacaagctc 180 gccgaggacgcagcacgcggcgtcgacggcggcggcggcaaggcagtgatcttcaacacc 240 atcaccatttccgacggcatctcgatgggcaccgaaggaatgaaatactccctcgtgtcg 300 cgggaagtcatcgccgactcgatcgaaaccgtggtggcctgtcagggttatgacggcgtg 360 gtcgccatcggcggctgcgacaagaacatgcccggctgcctgatcgccctcgcccgcctc 420 aaccgtccggcggtgttcgtctatggcggcaccatcctgccgggctgccacgacggcaag 480 aagctggacgtggtgtcggtgttcgaagcggtcggcgcccgcgccaaccaccgcatcgac 540 gatgccgaactgcacgccatcgaatccaatgccatccccggtccgggctcctgcggtggc 600 atgtataccgccaacaccatggcctccgccatcgaggcattagggatgagcctgccgggc 660 agttcggcccaggtggccatttcccgcgccaaggaactggattgcgagcgggccggcgcg 720 caggtcctcaagctcctggacctggggctcaaaccccgcgacatcatgaccaagaaggcg 780 ttcgagaacgccatcacggtggtgatcgccctgggcggctccaccaacgccgtgctgcac 840 ctcctggccatggccaacgcctgcggcgtcgacctgaagctcgacgatttcacccgcatc 900 gggcgcaaagtgccgatgctggcggatctgaaacccagcggcagatactccatggccgaa 960 ctggtggaaatcggcggcatccagccgctgatgaagaccttgctggacgcgggactcctg 1020 cacggcgactgcatgaccgtaaccggcaagaccctggaagaaaacctggccgacgcgccc 1080 gactacccggccggacaagacatgatccggtcgctggacaaccccatcaaaaaggacagc 1140 catctggtgatcctcaagggcaacctggcgccggaaggcgcggtcgccaagatcaccggc 1200 aaggaaggactgagcttcaccggcaccgcccgcgtattcgactgcgaggaagcggcgctc 1260 acggccatcctcgacggcacgatcgtgaaaggcgacgtcatcgtcatccgctatgaaggc 1320 cccaagggcggccccggcatgcgcgagatgctctcgccgacctcggcggtcatgggcaag 1380 ggattgggcaaggaggtcgccctcatcaccgacggccgcttttccggcggcacccacggc 1440 ttcgtggtcggccacatcacgccggaagcctacaccggcggccccctggcgatcgtccgg 1500 gacggcgataccatcaccatcgacgccgagacccgcgaattgagcctgcacgtcaccgac 1560 gatgaaatcggccggcgcctggcgcagtggactcaaccggcgccgcgctacaccaagggc 1620 gtgctggccaaatacgccaggttggtgagcccggcctcggaaggcgccgtcaccgacgac 1680 ggcctctga 1689 3-6 Sequences 3-6-1 SequenceNumber[ID] 6 3-6-2 MoleculeType AA 3-6-3 Length 562 source1...562 3-6-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-6-5 Residues MTDKHPRPHSSQVVDGMERAPSRAMLHAVGFADADFAKPQIGIASTWAMVTPCNMHINKL 60 AEDAARGVDGGGGKAVIFNTITISDGISMGTEGMKYSLVSREVIADSIETVVACQGYDGV 120 VAIGGCDKNMPGCLIALARLNRPAVFVYGGTILPGCHDGKKLDVVSVFEAVGARANHRID 180 DAELHAIESNAIPGPGSCGGMYTANTMASAIEALGMSLPGSSAQVAISRAKELDCERAGA 240 QVLKLLDLGLKPRDIMTKKAFENAITVVIALGGSTNAVLHLLAMANACGVDLKLDDFTRI 300 GRKVPMLADLKPSGRYSMAELVEIGGIQPLMKTLLDAGLLHGDCMTVTGKTLEENLADAP 360 DYPAGQDMIRSLDNPIKKDSHLVILKGNLAPEGAVAKITGKEGLSFTGTARVFDCEEAAL 420 TAILDGTIVKGDVIVIRYEGPKGGPGMREMLSPTSAVMGKGLGKEVALITDGRFSGGTHG 480 FVVGHITPEAYTGGPLAIVRDGDTITIDAETRELSLHVTDDEIGRRLAQWTQPAPRYTKG 540 VLAKYARLVSPASEGAVTDDGL 562 3-7 Sequences 3-7-1 SequenceNumber[ID] 7 3-7-2 MoleculeType DNA 3-7-3 Length 1650 source1...1650 3-7-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-7-5 Residues atgggcacggttgagcctggcgctatcggacaacatctgctcgcctgcctttaccaggcg 60 ggcgtcgggcacatcttcggcgttcccggcgattacgtgctgggcttctatgatctgatg 120 gccaaaggtcccgtccggcatatcgggaccacgcgggaggacaccgccgccttcgccgcc 180 gacggctatgcccgctgccggggcatgggcgcgctggcggtgacttacggggtcggtgcg 240 ctcaacaccgtcaacgccgtcgccggcgcctatgcggaatcctcgccggtggtggtcatc 300 agcggtgcgccgggggtgcgcgagcaaagggaagacccgttgatccaccaccgcttcggg 360 ccgttccggttccagcgcgagatattcgaacggatcacctgcgccgccgtggtgctggac 420 gatccggtgatcgccttccggcaggtggagcgtgcgctcgcggccgcccgtcagcactgc 480 aagccggtgtacatcgagattcccgccgaccgggtgatggcgccgggatatccgattcca 540 caggaaaccccggaaacgccttccagcgacgattcggccctggcggaggcggtcgccgag 600 gccgcggagctcctgggccgtgcggtgtcgccggtgatccttgcaggcgtcgagttgcac 660 cggcgagggctccaggacgccctcgtcggcctcgtcgagcaggcgcgcctgccggtggcg 720 gcgaccttgaccggcaagtcggtgttcgccgagcgccatcccgcctatctgggggtgtac 780 gagggtgcgatgagcacggaaaacgcgcgctacatggtcgagcagtccgacctcctgctg 840 atgctcggggtcacgctgaacgatgtcgacacgggcatctacacggcgcgtctcgatccg 900 cagcgcatcgtccgcgcagcccagaacgaggtcgtgattcgccatcaccgctatccccgc 960 gtcctgctcgcggacttcgtcacggccctggcgcggtccgtcaaggcccggggcgaggcg 1020 tttccgatgccggcggggccggaaccgtgggactttcccgcgccggaccggccgatgacg 1080 atcgcccggctggtggagcggctcgaccgcgcgctgacctccgacatgatcgtagtgtgc 1140 gacgtcggcgactgcctgttcgcagccaccgacctgcgcgtgcacgagcgcagcgaattc 1200 ctggcgtccgccttctatacctcgatggggttcgcggtgcccgccgccctcggggcccag 1260 atcgcccgtccggaccaccgggcgctgatcctggtcggcgacggtgccttccagatgacc 1320 ggaacggagctgtcgacccatgcccgtctcggcctggcgcccatcgtggtggtgctcgac 1380 aatcgcggttacagcaccgagcgcttcatcctcgacggagccttcaacgacatcgccgac 1440 tggcgcttccaccggctgggcgaggtgttcggccccctacagggctacgacgcgcccgac 1500 gaagcggcgttcgaaaacgcgctcagcgaagcgctggtcaaccgaaacatgccgagcctc 1560 atcaacgtccgtctttcccccggcgatgcctcgatagccatgaagcgtctcgccgggcat 1620 ctgcagtgccgggtcaagggcgagggctga 1650 3-8 Sequences 3-8-1 SequenceNumber[ID] 8 3-8-2 MoleculeType AA 3-8-3 Length 549 3-8-4-1 source1...549 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-8-5 Residues MGTVEPGAIGQHLLACLYQAGVGHIFGVPGDYVLGFYDLMAKGPVRHIGTTREDTAAFAA 60 DGYARCRGMGALAVTYGVGALNTVNAVAGAYAESSPVVVISGAPGVREQREDPLIHHRFG 120 PFRFQREIFERITCAAVVLDDPVIAFRQVERALAAARQHCKPVYIEIPADRVMAPGYPIP 180 QETPETPSSDDSALAEAVAEAAELLGRAVSPVILAGVELHRRGLQDALVGLVEQARLPVA 240 ATLTGKSVFAERHPAYLGVYEGAMSTENARYMVEQSDLLLMLGVTLNDVDTGIYTARLDP 300 QRIVRAAQNEVVIRHHRYPRVLLADFVTALARSVKARGEAFPMPAGPEPWDFPAPDRPMT 360 IARLVERLDRALTSDMIVVCDVGDCLFAATDLRVHERSEFLASAFYTSMGFAVPAALGAQ 420 IARPDHRALILVGDGAFQMTGTELSTHARLGLAPIVVVLDNRGYSTERFILDGAFNDIAD 480 WRFHRLGEVFGPLQGYDAPDEAAFENALSEALVNRNMPSLINVRLSPGDASIAMKRLAGH 540 LQCRVKGEG 549 3-9 Sequences 3-9-1 SequenceNumber[ID] 9 3-9-2 MoleculeType DNA 3-9-3 Length 1083 3-9-4-1 source1...1083 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Saccharomycescerevisiae NonEnglishQualifierValue 3-9-5 Residues atgtcttatcctgagaaatttgaaggtatcgctattcaatcacacgaagattggaaaaac 60 ccaaagaagacaaagtatgacccaaaaccattttacgatcatgacattgacattaagatc 120 gaagcatgtggtgtctgcggtagtgatattcattgtgcagctggtcattggggcaatatg 180 aagatgccgctagtcgttggtcatgaaatcgttggtaaagttgtcaagctagggcccaag 240 tcaaacagtgggttgaaagtcggtcaacgtgttggtgtaggtgctcaagtcttttcatgc 300 ttggaatgtgaccgttgtaagaatgataatgaaccatactgcaccaagtttgttaccaca 360 tacagtcagccttatgaagacggctatgtgtcgcagggtggctatgcaaactacgtcaga 420 gttcatgaacattttgtggtgcctatcccagagaatattccatcacatttggctgctcca 480 ctattatgtggtggtttgactgtgtactctccattggttcgtaacggttgcggtccaggt 540 aaaaaagttggtatagttggtcttggtggtatcggcagtatgggtacattgatttccaaa 600 gccatgggggcagagacgtatgttatttctcgttcttcgagaaaaagagaagatgcaatg 660 aagatgggcgccgatcactacattgctacattagaagaaggtgattggggtgaaaagtac 720 tttgacaccttcgacctgattgtagtctgtgcttcctcccttaccgacattgacttcaac 780 attatgccaaaggctatgaaggttggtggtagaattgtctcaatctctataccagaacaa 840 cacgaaatgttatcgctaaagccatatggcttaaaggctgtctccatttcttacagtgct 900 ttaggttccatcaaagaattgaaccaactcttgaaattagtctctgaaaaagatatcaaa 960 atttgggtggaaacattacctgttggtgaagccggcgtccatgaagccttcgaaaggatg 1020 gaaaagggtgacgttagatatagatttaccttagtcggctacgacaaagaattttcagac 1080 tag 1083 3-10 Sequences 3-10-1 SequenceNumber[ID] 10 3-10-2 MoleculeType AA 3-10-3 Length 360 source1...360 3-10-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Saccharomycescerevisiae NonEnglishQualifierValue 3-10-5 Residues MSYPEKFEGIAIQSHEDWKNPKKTKYDPKPFYDHDIDIKIEACGVCGSDIHCAAGHWGNM 60 KMPLVVGHEIVGKVVKLGPKSNSGLKVGQRVGVGAQVFSCLECDRCKNDNEPYCTKFVTT 120 YSQPYEDGYVSQGGYANYVRVHEHFVVPIPENIPSHLAAPLLCGGLTVYSPLVRNGCGPG 180 KKVGIVGLGGIGSMGTLISKAMGAETYVISRSSRKREDAMKMGADHYIATLEEGDWGEKY 240 FDTFDLIVVCASSLTDIDFNIMPKAMKVGGRIVSISIPEQHEMLSLKPYGLKAVSISYSA 300 LGSIKELNQLLKLVSEKDIKIWVETLPVGEAGVHEAFERMEKGDVRYRFTLVGYDKEFSD 360 3-11 Sequences 3-11-1 SequenceNumber[ID] 11 3-11-2 MoleculeType DNA 3-11-3 Length 783 source1...783 3-11-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-11-5 Residues atggcagcaacaaccattggtggtgcagctgcggcggaagcgccgctgctggacaagaag 60 tggctcacgttcgcactggcgatttacaccgtgttctacctgtgggtgcggtggtacgaa 120 ggtgtctatggctggtccgccggactggactcgttcgcgccggagttcgagacctactgg 180 atgaatttcctgtacaccgagatcgtcctggagatcgtgacggcttcgatcctgtggggc 240 tatctctggaagacccgcgaccgcaacctggccgcgctgaccccgcgtgaagagctgcgc 300 cgcaacttcacccacctggtgtggctggtggcctacgcctgggccatctactggggcgca 360 tcctacttcaccgagcaggacggcacctggcatcagacgatcgtgcgcgacaccgacttc 420 acgccgtcgcacatcatcgagttctatctgagctacccgatctacatcatcaccggtttt 480 gcggcgttcatctacgccaagacgcgtctgccgttcttcgcgaagggcatctcgctgccg 540 tacctggtgctggtggtgggtccgttcatgattctgccgaacgtgggtctgaacgaatgg 600 ggccacaccttctggttcatggaagagctgttcgtggcgccgctgcactacggcttcgtg 660 atcttcggctggctggcactggccgtcatgggcaccctgacccagaccttctacagcttc 720 gctcagggcgggctggggcagtcgctctgtgaagccgtggacgaaggcttgatcgcgaaa 780 taa 783 3-12 Sequences 3-12-1 SequenceNumber[ID] 12 3-12-2 MoleculeType AA 3-12-3 Length 260 source1...260 3-12-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-12-5 Residues MAATTIGGAAAAEAPLLDKKWLTFALAIYTVFYLWVRWYEGVYGWSAGLDSFAPEFETYW 60 MNFLYTEIVLEIVTASILWGYLWKTRDRNLAALTPREELRRNFTHLVWLVAYAWAIYWGA 120 SYFTEQDGTWHQTIVRDTDFTPSHIIEFYLSYPIYIITGFAAFIYAKTRLPFFAKGISLP 180 YLVLVVGPFMILPNVGLNEWGHTFWFMEELFVAPLHYGFVIFGWLALAVMGTLTQTFYSF 240 AQGGLGQSLCEAVDEGLIAK 260 3-13 Sequences 3-13-1 SequenceNumber[ID] 13 3-13-2 MoleculeType DNA 3-13-3 Length 744 source1...744 3-13-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-13-5 Residues atgagtgctgcgcaatctgcggttcgttcgcacgccgaagcggtccaggtatcccggacc 60 atcgactggatggcgttgttcgtggtgtttttcgtgatcgtgggctcgtaccacattcat 120 gccatgctcaccatgggtgactgggacttctggtcggactggaaagaccgtcgactgtgg 180 gtcacggtgaccccgatcgtactggtcaccttcccggcggccgtacaatcctacctgtgg 240 gagcggtatcgtctgccctggggagccaccgtgtgcgtcctgggtctgctgctgggcgag 300 tggatcaaccgttatttcaacttctggggctggacctacttcccgatcaacttcgtgttc 360 cctgcctcgctggtgccgggcgccatcatcctggacaccgtgctgatgctgtcgggcagc 420 tacctgttcaccgcgatcgtcggtgcgatgggctggggtctgatcttctacccgggcaac 480 tggccgatcatcgcgccgctgcacgtgccggtggaatacaacggcatgctgatgtcgatc 540 gccgacatccagggttacaactatgtgcgtacgggtacgcctgagtacatccgcatggta 600 gagaagggcaccctgcgtaccttcggtaaggacgtggcgccggtatcggcattcttctcc 660 gcgttcatgtcgatcctgatctacttcatgtggcacttcatcggtcgctggttctccaac 720 gaacggttcctgcagagcacctga 744 3-14 Sequences 3-14-1 SequenceNumer[ID] 14 3-14-2 MoleculeType AA 3-14-3 Length 247 source1...247 3-14-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-14-5 Residues MSAAQSAVRSHAEAVQVSRTIDWMALFVVFFVIVGSYHIHAMLTMGDWDFWSDWKDRRLW 60 VTVTPIVLVTFPAAVQSYLWERYRLPWGATVCVLGLLLGEWINRYFNFWGWTYFPINFVF 120 PASLVPGAIILDTVLMLSGSYLFTAIVGAMGWGLIFYPGNWPIIAPLHVPVEYNGMLMSI 180 ADIQGYNYVRTGTPEYIRMVEKGTLRTFGKDVAPVSAFFSAFMSILIYFMWHFIGRWFSN 240 ERFLQST 247 3-15 Sequences 3-15-1 SequenceNumer[ID] 15 3-15-2 MoleculeType DNA 3-15-3 Length 1245 source1...1245 3-15-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-15-5 Residues atgaaaacaataaaggaccggattgcaaaatggtctgcaatcggactgctgtccgccgtg 60 gcagcgaccgccttctatgcgccgagcgccagcgcccacggtgagaaatcgcaggccgcg 120 ttcatgcgtatgcgtaccatccactggtacgacctgagctggtcgaaagagaaagtcaag 180 atcaacgagaccgtggaaatcaaaggcaagttccacgtgttcgaaggctggccggaaacg 240 gtcgacgaaccggatgtggcgttcctgaacgtcggcatgccgggtccggtgttcatccgc 300 aaggaatcgtacatcggcggtcagctggtgccgcgttccgtacgtctggaaatcggcaag 360 acctatgacttccgggttgtcctcaaagcccgtcgtccgggtgactggcacgttcacacc 420 atgatgaacgtccagggcggtggaccgatcatcggtcccggcaaatggatcaccgtggaa 480 ggctccatgagtgaattccgcaaccccgtcaccaccctgaccggtcagacggtggacctg 540 gagaactacaacgaaggcaacacctatttctggcacgccttctggttcgccatcggagtt 600 gcctggatcggctactggtcgcgtcgaccgatcttcatcccccgtctgctgatggtggat 660 gccggtcgtgcggatgaactggtgtccgccaccgaccgcaaggtggcgatgggcttcctg 720 gccgccaccatcctgatcgtggtcatggccatgtccagcgccaacagcaagtacccgatc 780 accatcccgctgcaggccggcaccatgcgtggcatgaagccgctggaactgccggcgccg 840 acggtatcggtgaaagtggaagacgccacctaccgggtaccgggccgcgccatgcggatg 900 aagctgaccatcaccaaccacggcaacagcccgatccggctgggtgagttctacaccgcc 960 tcggtgcgtttcctggattccgacgtgtacaaggacaccaccggctatccggaagacctg 1020 ctggccgaagacggcctgagcgtcagcgacaacagcccgctggctccgggtgagacgcgc 1080 acggtcgacgtgacggcgtccgacgcggcgtgggaagtgtaccgtctgtccgacatcatc 1140 tacgatccggacagccgtttcgccggtctgctgttcttcttcgacgccactggcaaccgc 1200 caggtcgtccagatcgacgcaccgctgatcccgtcgttcatgtaa 1245 3-16 Sequences 3-16-1 SequenceNumber[ID] 16 3-16-2 MoleculeType AA 3-16-3 Length 414 source1...414 3-16-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-16-5 Residues MKTIKDRIAKWSAIGLLSAVAATAFYAPSASAHGEKSQAAFMRMRTIHWYDLSWSKEKVK 60 INETVEIKGKFHVFEGWPETVDEPDVAFLNVGMPGPVFIRKESYIGGQLVPRSVRLEIGK 120 TYDFRVVLKARRPGDWHVHTMMNVQGGGPIIGPGKWITVEGSMSEFRNPVTTLTGQTVDL 180 ENYNEGNTYFWHAFWFAIGVAWIGYWSRRPIFIPRLLMVDAGRADELVSATDRKVAMGFL 240 AATILIVVMAMSSANSKYPITIPLQAGTMRGMKPLELPAPTVSVKVEDATYRVPGRAMRM 300 KLTITNHGNSPIRLGEFYTASVRFLDSDVYKDTTGYPEDLLAEDGLSVSDNSPLAPGETR 360 TVDVTASDAAWEVYRLSDIIYDPDSRFAGLLFFFDATGNRQVVQIDAPLIPSFM 414 3-17 Sequences 3-17-1 SequenceNumber[ID] 17 3-17-2 MoleculeType DNA 3-17-3 Length 783 source1...783 3-17-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-17-5 Residues atggcagcaacaaccattggtggtgcagctgcggcggaagcgccgctgctggacaagaag 60 tggctcacgttcgcactggcgatttacaccgtgttctacctgtgggtgcggtggtacgaa 120 ggtgtctatggctggtccgccggactggactcgttcgcgccggagttcgagacctactgg 180 atgaatttcctgtacaccgagatcgtcctggagatcgtgacggcttcgatcctgtggggc 240 tatctctggaagacccgcgaccgcaacctggccgcgctgaccccgcgtgaagagctgcgc 300 cgcaacttcacccacctggtgtggctggtggcctacgcctgggccatctactggggcgca 360 tcctacttcaccgagcaggacggcacctggcatcagacgatcgtgcgcgacaccgacttc 420 acgccgtcgcacatcatcgagttctatctgagctacccgatctacatcatcaccggtttt 480 gcggcgttcatctacgccaagacgcgtctgccgttcttcgcgaagggcatctcgctgccg 540 tacctggtgctggtggtgggtccgttcatgattctgccgaacgtgggtctgaacgaatgg 600 ggccacaccttctggttcatggaagagctgttcgtggcgccgctgcactacggcttcgtg 660 atcttcggctggctggcactggccgtcatgggcaccctgacccagaccttctacagcttc 720 gctcagggcgggctggggcagtcgctctgtgaagccgtggacgaaggcttgatcgcgaaa 780 taa 783 3-18 Sequences 3-18-1 SequenceNumber[ID] 18 3-18-2 MoleculeType AA 3-18-3 Length 260 source1...260 3-18-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-18-5 Residues MAATTIGGAAAAEAPLLDKKWLTFALAIYTVFYLWVRWYEGVYGWSAGLDSFAPEFETYW 60 MNFLYTEIVLEIVTASILWGYLWKTRDRNLAALTPREELRRNFTHLVWLVAYAWAIYWGA 120 SYFTEQDGTWHQTIVRDTDFTPSHIIEFYLSYPIYIITGFAAFIYAKTRLPFFAKGISLP 180 YLVLVVGPFMILPNVGLNEWGHTFWFMEELFVAPLHYGFVIFGWLALAVMGTLTQTFYSF 240 AQGGLGQSLCEAVDEGLIAK 260 3-19 Sequences 3-19-1 SequenceNumber[ID] 19 3-19-2 MoleculeType DNA 3-19-3 Length 744 source1...744 3-19-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-19-5 Residues atgagtgctgcgcaatctgcggttcgttcgcacgccgaagcggtccaggtatcccggacc 60 atcgactggatggcgttgttcgtggtgtttttcgtgatcgtgggctcgtaccacattcac 120 gccatgctcaccatgggtgactgggacttctggtcggactggaaagaccgtcgactgtgg 180 gtcacggtgaccccgatcgtactggtcaccttcccggcggccgtacaatcctacctgtgg 240 gagcggtatcgtctgccctggggagccaccgtgtgcgtcctgggtctgctgctgggcgag 300 tggatcaaccgttatttcaacttctggggctggacctacttcccgatcaacttcgtgttc 360 cctgcctcgctggtgccgggcgccatcatcctggacaccgtgctgatgctgtcgggcagc 420 tacctgttcaccgcgatcgtcggtgcgatgggctggggtctgatcttctacccgggcaac 480 tggccgatcatcgcgccgctgcacgtgccggtggaatacaacggcatgctgatgtcgatc 540 gccgacatccagggttacaactatgtgcgtacgggtacgcctgagtacatccgcatggta 600 gagaagggcaccctgcgtaccttcggtaaggacgtggcgccggtatcggcattcttctcc 660 gcgttcatgtcgatcctgatctacttcatgtggcacttcatcggtcgctggttctccaac 720 gaacggttcctgcagagcacctga 744 3-20 Sequences 3-20-1 SequenceNumber[ID] 20 3-20-2 MoleculeType AA 3-20-3 Length 247 source1...247 3-20-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-20-5 Residues MSAAQSAVRSHAEAVQVSRTIDWMALFVVFFVIVGSYHIHAMLTMGDWDFWSDWKDRRLW 60 VTVTPIVLVTFPAAVQSYLWERYRLPWGATVCVLGLLLGEWINRYFNFWGWTYFPINFVE 120 PASLVPGAIILDTVLMLSGSYLFTAIVGAMGWGLIFYPGNWPIIAPLHVPVEYNGMLMSI 180 ADIQGYNYVRTGTPEYIRMVEKGTLRTFGKDVAPVSAFFSAFMSILIYFMWHFIGRWFSN 240 ERFLQST 247 3-21 Sequences 3-21-1 SequenceNumber[ID] 21 3-21-2 MoleculeType DNA 3-21-3 Length 1245 source1...1245 3-21-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-21-5 Residues atgaaaacaataaaggaccggattgcaaaatggtctgcaatcggactgctgtccgccgtg 60 gcagcgaccgccttctatgcgccgagcgccagcgcccacggtgagaaatcgcaggccgcg 120 ttcatgcgtatgcgtaccatccactggtacgacctgagctggtcgaaagagaaagtcaag 180 atcaacgagaccgtggaaatcaaaggcaagttccacgtgttcgaaggctggccggaaacg 240 gtcgacgaaccggatgtggcgttcctgaacgtcggcatgccgggtccggtgttcatccgc 300 aaggaatcgtacatcggcggtcagctggtgccgcgttccgtacgtctggaaatcggcaag 360 acctatgacttccgggttgtcctcaaagcccgtcgtccgggtgactggcacgttcacacc 420 atgatgaacgtccagggcggtggaccgatcatcggtcccggcaaatggatcaccgtggaa 480 ggctccatgagtgaattccgcaaccccgtcaccaccctgaccggtcagacggtggacctg 540 gagaactacaacgaaggcaacacctatttctggcacgccttctggttcgccatcggagtt 600 gcctggatcggctactggtcgcgtcgaccgatcttcatcccccgtctgctgatggtggat 660 gccggtcgtgcggacgaactggtgtccgccaccgaccgcaaggtggcgatgggcttcctg 720 gccgccaccatcctgatcgtggtcatggccatgtccagcgccaacagcaagtacccgatc 780 accatcccgctgcaggccggcaccatgcgtggcatgaagccgctggaactgccggcgccg 840 acggtatcggtgaaagtggaagacgccacctaccgggtaccgggccgcgccatgcggatg 900 aagctgaccatcaccaaccacggcaacagcccgatccggctgggtgagttctacaccgcc 960 tcggtgcgtttcctggattccgacgtgtacaaggacaccaccggctatccggaagacctg 1020 ctggccgaagacggcctgagcgtcagcgacaacagcccgctggctccgggtgagacccgc 1080 acggtcgacgtgacggcgtccgacgcggcgtgggaagtgtaccgtctgtccgacatcatc 1140 tacgatccggacagccgtttcgccggtctgctgttcttcttcgacgccactggcaaccgc 1200 caggtcgtccagatcgacgcaccgctgatcccgtcgttcatgtaa 1245 3-22 Sequences 3-22-1 SequenceNumber[ID] 22 3-22-2 MoleculeType AA 3-22-3 Length 414 source1...414 3-22-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-22-5 Residues MKTIKDRIAKWSAIGLLSAVAATAFYAPSASAHGEKSQAAFMRMRTIHWYDLSWSKEKVK 60 INETVEIKGKFHVFEGWPETVDEPDVAFLNVGMPGPVFIRKESYIGGQLVPRSVRLEIGK 120 TYDFRVVLKARRPGDWHVHTMMNVQGGGPIIGPGKWITVEGSMSEFRNPVTTLTGQTVDL 180 ENYNEGNTYFWHAFWFAIGVAWIGYWSRRPIFIPRLLMVDAGRADELVSATDRKVAMGFL 240 AATILIVVMAMSSANSKYPITIPLQAGTMRGMKPLELPAPTVSVKVEDATYRVPGRAMRM 300 KLTITNHGNSPIRLGEFYTASVRFLDSDVYKDTTGYPEDLLAEDGLSVSDNSPLAPGETR 360 TVDVTASDAAWEVYRLSDIIYDPDSRFAGLLFFFDATGNRQVVQIDAPLIPSFM 414 3-23 Sequences 3-23-1 SequenceNumber[ID] 23 3-23-2 MoleculeType DNA 3-23-3 Length 1584 source1...1584 3-23-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-23-5 Residues atggcacttagcaccgcaaccaaggccgcgacggacgcgctggctgccaatcgggcaccc 60 accagcgtgaatgcacaggaagtgcaccgttggctccagagcttcaactgggatttcaag 120 aacaaccggaccaagtacgccaccaagtacaagatggcgaacgagaccaaggaacagttc 180 aagctgatcgccaaggaatatgcgcgcatggaggcagtcaaggacgaaaggcagttcggt 240 agcctgcaggatgcgctgacccgcctcaacgccggtgttcgcgttcatccgaagtggaac 300 gagaccatgaaagtggtttcgaacttcctggaagtgggcgaatacaacgccatcgccgct 360 accgggatgctgtgggattccgcccaggcggcggaacagaagaacggctatctggcccag 420 gtgttggatgaaatccgccacacccaccagtgtgcctacgtcaactactacttcgcgaag 480 aacggccaggacccggccggtcacaacgatgctcgccgcacccgtaccatcggtccgctg 540 tggaagggcatgaagcgcgtgttttccgacggcttcatttccggcgacgccgtggaatgc 600 tccctcaacctgcagctggtgggtgaggcctgcttcaccaatccgctgatcgtcgcagtg 660 accgaatgggctgccgccaacggcgatgaaatcaccccgacggtgttcctgtcgatcgag 720 accgacgaactgcgccacatggccaacggttaccagaccgtcgtttccatcgccaacgat 780 ccggcttccgccaagtatctcaacacggacctgaacaacgccttctggacccagcagaag 840 tacttcacgccggtgttgggcatgctgttcgagtatggctccaagttcaaggtcgagccg 900 tgggtcaagacgtggaaccgctgggtgtacgaggactggggcggcatctggatcggccgt 960 ctgggcaagtacggggtggagtcgccgcgcagcctcaaggacgccaagcaggacgcttac 1020 tgggctcaccacgacctgtatctgctggcttatgcgctgtggccgaccggcttcttccgt 1080 ctggcgctgccggatcaggaagaaatggagtggttcgaggccaactaccccggctggtac 1140 gaccactacggcaagatctacgaggaatggcgcgcccgcggttgcgaggatccgtcctcg 1200 ggcttcatcccgctgatgtggttcatcgaaaacaaccatcccatctacatcgatcgcgtg 1260 tcgcaagtgccgttctgcccgagcttggccaagggcgccagcaccctgcgcgtgcacgag 1320 tacaacggccagatgcacaccttcagcgaccagtggggcgagcgcatgtggctggccgag 1380 ccggagcgctacgagtgccagaacatcttcgaacagtacgaaggacgcgaactgtcggaa 1440 gtgatcgccgaactgcacgggctgcgcagtgatggcaagaccctgatcgcccagccgcat 1500 gtccgtggcgacaagctgtggacgttggacgatatcaaacgcctgaactgcgtcttcaag 1560 aacccggtgaaggcattcaattga 1584 3-24 Sequences 3-24-1 SequenceNumber[ID] 24 3-24-2 MoleculeType AA 3-24-3 Length 527 source1...527 3-24-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-24-5 Residues MALSTATKAATDALAANRAPTSVNAQEVHRWLQSFNWDFKNNRTKYATKYKMANETKEQF 60 KLIAKEYARMEAVKDERQFGSLQDALTRLNAGVRVHPKWNETMKVVSNFLEVGEYNAIAA 120 TGMLWDSAQAAEQKNGYLAQVLDEIRHTHQCAYVNYYFAKNGQDPAGHNDARRTRTIGPL 180 WKGMKRVFSDGFISGDAVECSLNLQLVGEACFTNPLIVAVTEWAAANGDEITPTVFLSIE 240 TDELRHMANGYQTVVSIANDPASAKYLNTDLNNAFWTQQKYFTPVLGMLFEYGSKFKVEP 300 WVKTWNRWVYEDWGGIWIGRLGKYGVESPRSLKDAKQDAYWAHHDLYLLAYALWPTGFFR 360 LALPDQEEMEWFEANYPGWYDHYGKIYEEWRARGCEDPSSGFIPLMWFIENNHPIYIDRV 420 SQVPFCPSLAKGASTLRVHEYNGQMHTFSDQWGERMWLAEPERYECQNIFEQYEGRELSE 480 VIAELHGLRSDGKTLIAQPHVRGDKLWTLDDIKRLNCVFKNPVKAFN 527 3-25 Sequences 3-25-1 SequenceNumber[ID] 25 3-25-2 MoleculeType DNA 3-25-3 Length 1170 source1...1170 3-25-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-25-5 Residues atgagcatgttaggagaaagacgccgcggtctgaccgatccggaaatggcggccgtcatt 60 ttgaaggcgcttcctgaagctccgctggacggcaacaacaagatgggttatttcgtcacc 120 ccccgctggaaacgcttgacggaatatgaagccctgaccgtttatgcgcagcccaacgcc 180 gactggatcgccggcggcctggactggggcgactggacccagaaattccacggcggccgc 240 ccttcctggggcaacgagaccacggagctgcgcaccgtcgactggttcaagcaccgtgac 300 ccgctccgccgttggcatgcgccgtacgtcaaggacaaggccgaggaatggcgctacacc 360 gaccgcttcctgcagggttactccgccgacggtcagatccgggcgatgaacccgacctgg 420 cgggacgagttcatcaaccggtattggggcgccttcctgttcaacgaatacggattgttc 480 aacgctcattcgcagggcgcccgggaggcgctgtcggacgtaacccgcgtcagcctggct 540 ttctggggcttcgacaagatcgacatcgcccagatgatccaactcgaacggggtttcctc 600 gccaagatcgtacccggtttcgacgagtccacagcggtgccgaaggccgaatggacgaac 660 ggggaggtctacaagagcgcccgtctggccgtggaagggctgtggcaggaggtgttcgac 720 tggaacgagagcgctttctcggtgcacgccgtctatgacgcgctgttcggtcagttcgtc 780 cgccgcgagttctttcagcggctggctccccgcttcggcgacaatctgacgccattcttc 840 atcaaccaggcccagacatacttccagatcgccaagcagggcgtacaggatctgtattac 900 aactgtctgggtgacgatccggagttcagcgattacaaccgtaccgtgatgcgcaactgg 960 accggcaagtggctggagcccacgatcgccgctctgcgcgacttcatggggctgtttgcg 1020 aagctgccggcgggcaccactgacaaggaagaaatcaccgcgtccctgtaccgggtggtc 1080 gacgactggatcgaggactacgccagcaggatcgacttcaaggcggaccgcgatcagatc 1140 gttaaagcggttctggcaggattgaaataa 1170 3-26 Sequences 3-26-1 SequenceNumber[ID] 26 3-26-2 MoleculeType AA 3-26-3 Length 389 source1...389 3-26-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-26-5 Residues MSMLGERRRGLTDPEMAAVILKALPEAPLDGNNKMGYFVTPRWKRLTEYEALTVYAQPNA 60 DWIAGGLDWGDWTQKFHGGRPSWGNETTELRTVDWFKHRDPLRRWHAPYVKDKAEEWRYT 120 DRFLQGYSADGQIRAMNPTWRDEFINRYWGAFLFNEYGLFNAHSQGAREALSDVTRVSLA 180 FWGFDKIDIAQMIQLERGFLAKIVPGFDESTAVPKAEWTNGEVYKSARLAVEGLWQEVFD 240 WNESAFSVHAVYDALFGQFVRREFFQRLAPRFGDNLTPFFINQAQTYFQIAKQGVQDLYY 300 NCLGDDPEFSDYNRTVMRNWTGKWLEPTIAALRDFMGLFAKLPAGTTDKEEITASLYRVV 360 DDWIEDYASRIDFKADRDQIVKAVLAGLK 389 3-27 Sequences 3-27-1 SequenceNumber[ID] 27 3-27-2 MoleculeType DNA 3-27-3 Length 426 source1...426 3-27-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue Residues 3-27-5 atgagcgtaaacagcaacgcatacgacgccggcatcatgggcctgaaaggcaaggacttc 60 gccgatcagttctttgccgacgaaaaccaagtggtccatgaaagcgacacggtcgttctg 120 gtcctcaagaagtcggacgagatcaatacctttatcgaggagatccttctgacggactac 180 aagaagaacgtcaatccgacggtaaacgtggaagaccgcgcgggttactggtggatcaag 240 gccaacggcaagatcgaggtcgattgcgacgagatttccgagctgttggggcggcagttc 300 aacgtctacgacttcctcgtcgacgtttcctccaccatcggccgggcctataccctgggc 360 aacaagttcaccattaccagtgagctgatgggcctggaccgcaagctcgaagactatcac 420 gcttaa 426 3-28 Sequences 3-28-1 SequenceNumber[ID] 28 3-28-2 MoleculeType AA 3-28-3 Length 138 source1...138 3-28-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-28-5 Residues MSVNSNAYDAGIMGLKGKDFADQFFADENQVVHESDTVVLVLKKSDEINTFIEEILLTDY 60 KKNVNPTVNVEDRAGYWWIKANGKIEVDCDEISELLGRQFNVYDFLVDVSSTIGRAYTLG 120 NKFTITSELMGLDRKLED 138 3-29 Sequences 3-29-1 SequenceNumber[ID] 29 3-29-2 MoleculeType DNA 3-29-3 Length 513 source1...513 3-29-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-29-5 Residues atggcgaaactgggtatacacagcaacgacacccgcgacgcctgggtgaacaagatcgcg 60 cagctcaacaccctggaaaaagcggccgagatgctgaagcagttccggatggaccacacc 120 acgccgttccgcaacagctacgaactggacaacgactacctctggatcgaggccaagctc 180 gaagagaaggtcgccgtcctcaaggcacgcgccttcaacgaggtggacttccgtcataag 240 accgctttcggcgaggatgccaagtccgttctggacggcaccgtcgcgaagatgaacgcg 300 gccaaggacaagtgggaggcggagaagatccatatcggtttccgccaggcctacaagccg 360 ccgatcatgccggtgaactatttcctggacggcgagcgtcagttggggacccggctgatg 420 gaactgcgcaacctcaactactacgacacgccgctggaagaactgcgcaaacagcgcggt 480 gtgcgggtggtgcatctgcagtcgccgcactga 513 3-30 Sequences 3-30-1 SequenceNumber[ID] 30 3-30-2 MoleculeType AA 3-30-3 Length 170 source1..170 3-30-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-30-5 Residues MAKLGIHSNDTRDAWVNKIAQLNTLEKAAEMLKQFRMDHTTPFRNSYELDNDYLWIEAKL 60 EEKVAVLKARAFNEVDFRHKTAFGEDAKSVLDGTVAKMNAAKDKWEAEKIHIGFRQAYKP 120 PIMPVNYFLDGERQLGTRLMELRNLNYYDTPLEELRKQRGVRVVHLQSPH 170 3-31 Sequences 3-31-1 SequenceNumber[ID] 31 3-31-2 MoleculeType DNA 3-31-3 Length 312 source1...312 3-31-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-31-5 Residues atggtcgaatcggcatttcagccattttcgggcgacgcagacgaatggttcgaggaacca 60 cggccccaggccggtttcttcccttccgcggactggcatctgctcaaacgggacgagacc 120 tacgcagcctatgccaaggatctcgatttcatgtggcggtgggtcatcgtccgggaagaa 180 aggatcgtccaggagggttgctcgatcagcctggagtcgtcgatccgcgccgtgacgcac 240 gtactgaattattttggtatgaccgaacaacgcgccccggcagaggaccggaccggcgga 300 gttcaacattga 312 3-32 Sequences 3-32-1 SequenceNumber[ID] 32 3-32-2 MoleculeType AA 3-32-3 Length 103 source1...103 3-32-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-32-5 Residues MVESAFQPFSGDADEWFEEPRPQAGFFPSADWHLLKRDETYAAYAKDLDFMWRWVIVREE 60 RIVQEGCSISLESSIRAVTHVLNYFGMTEQRAPAEDRTGGVQH 103 3-33 Sequences 3-33-1 SequenceNumber[ID] 33 3-33-2 MoleculeType DNA 3-33-3 Length 1047 source1...1047 3-33-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-33-5 Residues atgcagcgagttcacactatcacggcggtgacggaggatggcgaatcgctccgcttcgaa 60 tgccgttcggacgaggacgtcatcaccgccgccctgcgccagaacatctttctgatgtcg 120 tcctgccgggagggcggctgtgcgacctgcaaggccttgtgcagcgaaggggactacgac 180 ctcaagggctgcagcgttcaggcgctgccgccggaagaggaggaggaagggttggtgttg 240 ttgtgccggacctacccgaagaccgacctggaaatcgaactgccctatacccattgccgc 300 atcagttttggtgaggtcggcagtttcgaggcggaggtcgtcggcctcaactgggtttcg 360 agcaacaccgtccagtttcttttgcagaagcggcccgacgagtgcggcaaccgtggcgtg 420 aaattcgaacccggtcagttcatggacctgaccatccccggcaccgatgtctcccgctcc 480 tactcgccggcgaaccttcctaatcccgaaggccgcctggagttcctgatccgcgtgtta 540 ccggagggacggttttcggactacctgcgcaatgacgcgcgtgtcggacaggtcctctcg 600 gtcaaagggccactgggcgtgttcggtctcaaggagcggggcatggcgccgcgctatttc 660 gtggccggcggcaccgggttggcgccggtggtctcgatggtgcggcagatgcaggagtgg 720 accgcgccgaacgagacccgcatctatttcggtgtgaacaccgagccggaattgttctac 780 atcgacgagctcaaatccctggaacgatcgatgcgcaatctcaccgtgaaggcctgtgtc 840 tggcacccgagcggggactgggaaggcgagcagggctcgcccatcgatgcgttgcgggaa 900 gacctggagtcctccgacgccaacccggacatttatttgtgcggtccgccgggcatgatc 960 gatgccgcctgcgagctggtacgcagccgcggtatccccggcgaacaggtcttcttcgaa 1020 aaattcctgccgtccggggcggcctga 1047 3-34 Sequences 3-34-1 SequenceNumber[ID] 34 3-34-2 MoleculeType AA 3-34-3 Length 348 source1...348 3-34-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-34-5 Residues MQRVHTITAVTEDGESLRFECRSDEDVITAALRQNIFLMSSCREGGCATCKALCSEGDYD 60 LKGCSVQALPPEEEEEGLVLLCRTYPKTDLEIELPYTHCRISFGEVGSFEAEVVGLNWVS 120 SNTVQFLLQKRPDECGNRGVKFEPGQFMDLTIPGTDVSRSYSPANLPNPEGRLEFLIRVL 180 PEGRFSDYLRNDARVGQVLSVKGPLGVFGLKERGMAPRYFVAGGTGLAPVVSMVRQMQEW 240 TAPNETRIYFGVNTEPELFYIDELKSLERSMRNLTVKACVWHPSGDWEGEQGSPIDALRE 300 DLESSDANPDIYLCGPPGMIDAACELVRSRGIPGEQVFFEKFLPSGAA 348 3-35 Sequences 3-35-1 SequenceNumber[ID] 35 3-35-2 MoleculeType DNA 3-35-3 Length 1806 source1...1806 3-35-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-35-5 Residues atgcaaatttgcaaactggcaagtggctgcggcgggtcgatgctggcgatggccgccgtg 60 ctagccgcgcaatccacgcacgccaattcggagctggaccggctgtcgaaggacgaccgg 120 aactgggtcatgcagaccaaggactacagcgccacccacttcagccggctgacggaaatc 180 aatagccacaacgtcaagaacctgaaggtggcctggaccctgtccaccggcacgttgcat 240 ggccacgaaggtgcgccgttggtggtggacggcatcatgtacatccacacgccgttcccc 300 aacaacgtctatgcagtcgacctgaacgacacccgcaagatgctgtggcagtacaagccc 360 aagcagaatccggcggcccgcgcggtggcttgctgcgacgtggtcaaccgcggtctggcc 420 tacgtgccggccggcgagcacggtccggcgaagatcttcctcaaccagcttgacggccac 480 atcgtcgcactcaacgccaagaccggcgaagagatatggaagatggaaaattccgacatc 540 gccatgggctccaccctcaccggcgcgcctttcgtggtgaaggacaaggtactggtaggt 600 tcggccggggccgagctgggcgtgcgtggctacgtcacggcctataacatcaaggacggc 660 aagcaggagtggcgggcctatgccaccggtcccgacgaagacttgttgctggacaaggac 720 ttcaacaaggacaacccgcattacggtcagttcggcctggggctctcaacctgggagggt 780 gatgcctggaagatcggcggcggcaccaattggggctggtatgcctatgatcccaagttg 840 gacatgatctactacggttccggcaatccggcaccctggaacgagaccatgcggcccggc 900 gacaacaaatggaccatgaccatctggggccgcgacgccgacaccggccgcgccaagttc 960 ggctaccagaagacgccgcacgacgagtgggattacgccggtgtcaactacatgggtctg 1020 tccgaacaggaagtggacggcaagctgacgccgctgctgacccatcccgaccgcaacggt 1080 ctggtgtatacgctgaaccgggaaaccggcgccctggtcaatgccttcaagatcgatgac 1140 accgtcaactgggtgaaaaaggtcgatctgaagaccggcctgccgatccgcgatccggag 1200 tacagcacccgcatggaccacaatgccaaaggcatctgtccctcggccatgggctatcac 1260 aaccagggcatcgagtcctacgatccggacaagaagctgttcttcatgggcgtgaaccac 1320 atctgcatggactgggagccgttcatgctgccctaccgcgccggccagttctttgtgggg 1380 gcgaccctcaacatgtatccgggacccaaggggatgctgggtcaggtcaaggcgatgaac 1440 gcggtcaccggcaagatggaatgggaagtgccggagaagtttgcggtctggggtggcacc 1500 ttggcgaccgccggcgacctcgtgttctacggtaccctcgacggcttcatcaaggcccgc 1560 gacacccgtaccggcgagctgaagtggcagttccagttgccctccggcgtgatcggccat 1620 cccatcacctatcagcacaacggcaagcaatacattgccatctactccggcgtcggcggc 1680 tggccaggagtagggctggtattcgacctgaaggacccgaccgcaggtctgggagctgtg 1740 ggtgcgttcagggaactggcgcattacacccagatgggtggatcggtgttcgtgttctcg 1800 ctttga 1806 3-36 Sequences 3-36-1 SequenceNumber[ID] 36 3-36-2 MoleculeType AA 3-36-3 Length 601 source1...601 3-36-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-36-5 Residues MQICKLASGCGGSMLAMAAVLAAQSTHANSELDRLSKDDRNWVMQTKDYSATHFSRLTEI 60 NSHNVKNLKVAWTLSTGTLHGHEGAPLVVDGIMYIHTPFPNNVYAVDLNDTRKMLWQYKP 120 KQNPAARAVACCDVVNRGLAYVPAGEHGPAKIFLNQLDGHIVALNAKTGEEIWKMENSDI 180 AMGSTLTGAPFVVKDKVLVGSAGAELGVRGYVTAYNIKDGKQEWRAYATGPDEDLLLDKD 240 FNKDNPHYGQFGLGLSTWEGDAWKIGGGTNWGWYAYDPKLDMIYYGSGNPAPWNETMRPG 300 DNKWTMTIWGRDADTGRAKFGYQKTPHDEWDYAGVNYMGLSEQEVDGKLTPLLTHPDRNG 360 LVYTLNRETGALVNAFKIDDTVNWVKKVDLKTGLPIRDPEYSTRMDHNAKGICPSAMGYH 420 NQGIESYDPDKKLFFMGVNHICMDWEPFMLPYRAGQFFVGATLNMYPGPKGMLGQVKAMN 480 AVTGKMEWEVPEKFAVWGGTLATAGDLVFYGTLDGFIKARDTRTGELKWQFQLPSGVIGH 540 PITYQHNGKQYIAIYSGVGGWPGVGLVFDLKDPTAGLGAVGAFRELAHYTQMGGSVFVFS 600 L 601 3-37 Sequences 3-37-1 SequenceNumber[ID] 37 3-37-2 MoleculeType DNA 3-37-3 Length 573 source1...573 3-37-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-37-5 Residues atgctcaccagcagtccttattaccggtccggctacgtattcgtctaccgcaaggacacg 60 ggactgagcatccaagattggaacagcgcggcactgaagaccgtgaagcggatcgcattc 120 atgccggataccccggctgagacgatgatccgcaccatcggccgctacaacgacatgttc 180 aactacatgcactctctggtcggtttcaagtcgcggcgtaaccagtacgtgcgctacgac 240 ccggccaagctggtggcggaagtcgccgacggcaacgcggaagtcgcggtgttgtggggg 300 ccggcggcggcgcgctatgtcagaggggcggggctggccatgaccgtcatccccgacgac 360 aaccggcggtccgacggcgagaaagtgccccaccactattcgacttccgtcggcgtgcgc 420 aagggcgaggaggccctgctcaagcagatcgaccaggttctggcccgcttcggcaaggaa 480 gtgaatgcggtgctggaggcggaaggcattccgctgttgcccatggatgaaaaaccggcc 540 aggacggcttcccatgatcgaaggaaaggctag 573 3-38 Sequences 3-38-1 SequenceNumber[ID] 38 3-38-2 MoleculeType AA 3-38-3 Length 190 source1...190 3-38-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-38-5 Residues MLTSSPYYRSGYVFVYRKDTGLSIQDWNSAALKTVKRIAFMPDTPAETMIRTIGRYNDMF 60 NYMHSLVGFKSRRNQYVRYDPAKLVAEVADGNAEVAVLWGPAAARYVRGAGLAMTVIPDD 120 NRRSDGEKVPHHYSTSVGVRKGEEALLKQIDQVLARFGKEVNAVLEAEGIPLLPMDEKPA 180 RTASHDRRKG 190 3-39 Sequences 3-39-1 SequenceNumber[ID] 39 3-39-2 MoleculeType DNA 3-39-3 Length 480 source1...480 3-39-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-39-5 Residues atgaagctgaagaatgcgaggttcgacgtggctggcatgtgtgtcgccgggttgttggcg 60 ctgcccgcgcaggccgacattaccctgcggcatgccgtcaccggcgagacgctggagttg 120 tcctacgccaaggcgggcggcgacacgcaagccgtcaagcagttcctgcagaccggcaag 180 aacccttacaacggcaacaaggaggtagtggaacagggacatagtctgtatctgtcagcc 240 tgttccggctgccacggccatgaggccgaaggcaagctcggtccgggattggcggacgac 300 tattggacctatccccgcgcggccaccgacgtcggtttgttcgaaatcctgttcggcggc 360 gcgcagggcatgatggggccgcagtacgtcaacctcaacaatgacgaaatgctcaagatc 420 atggcctggatccgcagcctttaccggggcgatccagccaaggccgaatggctgaaatga 480 3-40 Sequences 3-40-1 SequenceNumber[ID] 40 3-40-2 MoleculeType AA 3-40-3 Length 159 source1...159 3-40-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-40-5 Residues MKLKNARFDVAGMCVAGLLALPAQADITLRHAVTGETLELSYAKAGGDTQAVKQFLQTGK 60 NPYNGNKEVVEQGHSLYLSACSGCHGHEAEGKLGPGLADDYWTYPRAATDVGLFEILFGG 120 AQGMMGPQYVNLNNDEMLKIMAWIRSLYRGDPAKAEWLK 159 3-41 Sequences 3-41-1 SequenceNumber[ID] 41 3-41-2 MoleculeType DNA 3-41-3 Length 285 source1...285 3-41-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-41-5 Residues atgatgcagaaaacgagtttcgtcgcggccgccatggccgtttcgttcgcggcgggtgtc 60 caggcctatgacggtacccactgcaaggcgcccggaaactgctgggagcccaagcccggt 120 tatccggacaaggtcgccggcagcaagtacgaccccaagcatgacccgaacgagctcaac 180 aagcaggcggagtcgatcaaggcgatggaagcccgcaaccagaagcgcgtggagaactac 240 gccaagaccggcaagttcgtctacaaggtcgaagacatcaaatga 285 3-42 Sequences 3-42-1 SequenceNumber[ID] 42 3-42-2 MoleculeType AA 3-42-3 Length 94 source1...94 3-42-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-42-5 Residues MMQKTSFVAAAMAVSFAAGVQAYDGTHCKAPGNCWEPKPGYPDKVAGSKYDPKHDPNELN 60 KQAESIKAMEARNQKRVENYAKTGKFVYKVEDIK 94 3-43 Sequences 3-43-1 SequenceNumber[ID] 43 3-43-2 MoleculeType DNA 3-43-3 Length 1020 source1...1020 3-43-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-43-5 Residues atgaatctagacaccccccttgccgacggcctggaacgcgccaaacgcttcgaacagggg 60 ctgcagcaagtcgtgctcgggcaggagcgcccgatccgcctgctgaccctggccgtgttc 120 gcccgcggtcatgcgctgctcgaaggcggcgtcggcgtcgggaagaccaccttgctccgt 180 gcggtggcgcgcggcatcggcggcgattacgagcggatcgagggcaccatcgacctgatg 240 ccgaacgatctggtctattacacctacctggacgagcaaggtaggccgggcgtcgcgccg 300 gggcctttgctcaagcacggggagcagctttccatttttttcttcaacgagatcaaccgc 360 gcccggccccaggtgcattccctcctgctacgggtcatggccgagcgcagcgtgtcggct 420 ttcaaccgcgagtaccggtttccgtacctgcaggtgttcgccgaccgcaaccgggtggaa 480 aaggaggagactttcgaattgcccgcggcggcgcgcgaccgcttcatgctcgaaatcgcc 540 atcgagccgccggccgatcctgcgcatatcgaccaaatcctgttcgacccgcgtttctac 600 gatcccgaccggctggtcgcgtccgcgccggccgatacgctctcgttccgtgaactcaac 660 ggcattgccgaagccctgcaaggcggcatccacgtcagcgcccgtctcagatcctatgtc 720 caggatctgtggcgcgcgacccggcggccggaggatttcggcatcgctctccacgaggcg 780 gattccggcgacatgatcgaggccggttccagtccccgcggcatgagctacttggtccgg 840 ctggcgcgggtgcaggcgtggctcagtggccgggaccgggtcgagccggaggacgttcaa 900 tacgtgttcgctccggcggtcggccaccgcatcttcctcaagccggtctacgaataccgc 960 cgcgccgagctgatcccggagctggtcggcaagctgatccgccggatcgcggcgccatga 1020 3-44 Sequences 3-44-1 SequenceNumber[ID] 44 3-44-2 MoleculeType AA 3-44-3 Length 339 source1...339 3-44-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-44-5 Residues MNLDTPLADGLERAKRFEQGLQQVVLGQERPIRLLTLAVFARGHALLEGGVGVGKTTLLR 60 AVARGIGGDYERIEGTIDLMPNDLVYYTYLDEQGRPGVAPGPLLKHGEQLSIFFFNEINR 120 ARPQVHSLLLRVMAERSVSAFNREYRFPYLQVFADRNRVEKEETFELPAAARDRFMLEIA 180 IEPPADPAHIDQILFDPRFYDPDRLVASAPADTLSFRELNGIAEALQGGIHVSARLRSYV 240 QDLWRATRRPEDFGIALHEADSGDMIEAGSSPRGMSYLVRLARVQAWLSGRDRVEPEDVQ 300 YVFAPAVGHRIFLKPVYEYRRAELIPELVGKLIRRIAAP 339 3-45 Sequences 3-45-1 SequenceNumber[ID] 45 3-45-2 MoleculeType DNA 3-45-3 Length 873 source1...873 3-45-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-45-5 Residues gtggtttggtctctcctgccggtcgcggccttggtatcggttccacttcatggcgccact 60 tcgctctcgttcgacacgccccgcgccttcggctacgtcatcggtgatctcatccgccac 120 gaggttcgggtcgaaaccgatgcggggcagggaatagaggctgcgtccctgcccaaggaa 180 ggctggatcaaccgctggctgctgctgcggcgggtcgaagtccgccgcgagggcaggcac 240 cggatactgacgctggaataccagactttctacgccccgttggaagtgaagaacctcacg 300 attcccggcttcgagctgcaactggccggttcgggcgaacggttggcggtcccggactgg 360 actttcaccaccgcgccgatccgggagctgtcggtgctgcgcgccgaaggcccgtcgatg 420 cgtccggacgccgcaccggcgccgctgccgactctcggccccgccgccgcgagcgtcggt 480 tccggcctcgcagccacgggcgcgctggcctggtgggcctatctgagcgcctggctgccg 540 ttcgtgtcgcgcggccgtcatttcgccgaggcccgccgggtgctgcgggatctgcgcggc 600 ctgggagacagccgggaggcattgcgcagaggtttttcctgtctgcaccaggctttcaat 660 cggacttcgggtgagccgctgttcatcgaagggctggacgagttcttccggagccatccg 720 gcctacgatctcttgcgggacgagatccaggacttcttcctggcctcgtatgaagtcttt 780 ttcggagagggcgcaccggcgccgtcgttcgacctggcgcgcatggaggcgttggcccgt 840 tcgtgccagcttgccgaaaggaggcggccatga 873 3-46 Sequences 3-46-1 SequenceNumber[ID] 46 3-46-2 MoleculeType AA 3-46-3 Length 290 source1...290 3-46-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-46-5 Residues VVWSLLPVAALVSVPLHGATSLSFDTPRAFGYVIGDLIRHEVRVETDAGQGIEAASLPKE 60 GWINRWLLLRRVEVRREGRHRILTLEYQTFYAPLEVKNLTIPGFELQLAGSGERLAVPDW 120 TFTTAPIRELSVLRAEGPSMRPDAAPAPLPTLGPAAASVGSGLAATGALAWWAYLSAWLP 180 FVSRGRHFAEARRVLRDLRGLGDSREALRRGFSCLHQAFNRTSGEPLFIEGLDEFFRSHP 240 AYDLLRDEIQDFFLASYEVFFGEGAPAPSFDLARMEALARSCQLAERRRP 290 3-47 Sequences 3-47-1 SequenceNumber[ID] 47 3-47-2 MoleculeType DNA 3-47-3 Length 984 source1...984 3-47-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-47-5 Residues atgaccgattgggcgctggacacgccgtacttgctgtggggcctgccgctggcgctgctt 60 ccgttgtggcggttgccgctgcgccctgccccgtgttcctggcatgcattgttgcccgcc 120 gatactgcgtcgcgggccgtcgacctgagtctgcgcctcgccggtgccggcgccatcctg 180 gcgctgctgctgggcagtgccggtctgcatcggcgcgagtacaccgtcgaacgcaccggc 240 tacggcgcccacatggtgctgctgctggaccgcagccgcagcatggatgacagcttcgca 300 gggcgtactcccacgggcggcgaggaatccaagtccgccgcggcggagcgcctcctgagc 360 ggtttcgtctcgagcggacgcaacgatctggtcggggtcgccgccttcagcacctccccg 420 ttgttcgtgctgccgctgaccgacaacaaggctgcggtgctggcggcggtccacgccatg 480 aagctgccgggtctggcgcagacgcatgtgagcaaggggctggcgatggcgctttcgtat 540 ttcggcgacgattcgaccgcgggttcgcgtatcgtcctgctggtgtccgacggtgccgcc 600 gaggtggacccggacagcgagctgaagctgcgccgctggttcaaggagaagggcgtacgg 660 ctgtactggatattcctgcgcaccgcgggcagccacggtatcttcgaaactccggacaac 720 ccggaggaagacaacgcccaggcgcggcccgagcgctatctgcatctgtttttcaacagt 780 ctgggcatcccctaccgcgcctacgaggcggaagacgccgacgccctcaagcgcgccatc 840 gccgacgtcgaccgcgaggagcagcggccgctgcgctatgccgagcgggtgccgcggcgg 900 gatctgcaagccttttgttatctggcggcggcgctggctctggcctggctggtcgccgcg 960 aagggcatggaggtggcgcgatga 984 3-48 Sequences 3-48-1 SequenceNumber[ID] 48 3-48-2 MoleculeType AA 3-48-3 Length 327 source1...327 3-48-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-48-5 Residues MTDWALDTPYLLWGLPLALLPLWRLPLRPAPCSWHALLPADTASRAVDLSLRLAGAGAIL 60 ALLLGSAGLHRREYTVERTGYGAHMVLLLDRSRSMDDSFAGRTPTGGEESKSAAAERLLS 120 GFVSSGRNDLVGVAAFSTSPLFVLPLTDNKAAVLAAVHAMKLPGLAQTHVSKGLAMALSY 180 FGDDSTAGSRIVLLVSDGAAEVDPDSELKLRRWFKEKGVRLYWIFLRTAGSHGIFETPDN 240 PEEDNAQARPERYLHLFFNSLGIPYRAYEAEDADALKRAIADVDREEQRPLRYAERVPRR 300 DLQAFCYLAAALALAWLVAAKGMEVAR 327 3-49 Sequences 3-49-1 SequenceNumber[ID] 49 3-49-2 MoleculeType DNA 3-49-3 Length 519 source1...519 3-49-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-49-5 Residues atgctggccttgtcggcgttgctggagctcaggcaatggcggaaggccgcggcggccaat 60 gccgatatcgccgagctgctgggggggcacgacatcgccccggaacggctggcggcggca 120 tcgccccaagtcctgttggcgcgggccgtgtatttcgtgcggcacgagcgctacggcgac 180 gcgctggagctgctgaacctgctggagacccggggcgatggcgccttccgcgccgacgtg 240 tattacaaccagggcaatctgcagcttgcccaggctctggaccgcgtcgaaaaatcggaa 300 atggaccaggcccgggtcttcgccgaactggccaaggaagcctaccggcgtgccttgtcg 360 ctggcacccggccactgggacgccaaatacaacctggaagtggccatgcgcctcatgccc 420 gaaatggaccgggtcagccctgccgatgacgaggcgcccgcggctgaatccaaacggctg 480 tggacaggtttgcccggactcccgcgaggcctgccttga 519 3-50 Sequences 3-50-1 SequenceNumber[ID] 50 3-50-2 MoleculeType AA 3-50-3 Length 172 source1...172 3-50-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-50-5 Residues MLALSALLELRQWRKAAAANADIAELLGGHDIAPERLAAASPQVLLARAVYFVRHERYGD 60 ALELLNLLETRGDGAFRADVYYNQGNLQLAQALDRVEKSEMDQARVFAELAKEAYRRALS 120 LAPGHWDAKYNLEVAMRLMPEMDRVSPADDEAPAAESKRLWTGLPGLPRGLP 172 3-51 Sequences 3-51-1 SequenceNumber[ID] 51 3-51-2 MoleculeType DNA 3-51-3 Length 972 source1...972 3-51-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-51-5 Residues ttgagcatctggcggcagcgcgttgccgatccggtttttgccggcctgattgtagccctc 60 cttctggcggtagccgcctgtttcccgctccggctggtgctggagcggctggtgttcagc 120 cacatcgtcgtcgtcgacatcacccgcagcatgaacgtcgaggactaccggcgaggcgcg 180 cgcgccgtgtcgcggctggaattcgtcaggcagagcctgatcggcgccgtggccgacctg 240 ccctgcggctccgctgtgggggtgggcgttttcaccgaacgcgagccggcgctactgttc 300 gagccgatcgaaacctgcgccggcttttccgccatcagcgccgccatcgaacagctcgac 360 tggcgcatggcctgggctgccgacagtctgatcgccgcaggtctgcacaacaccctggat 420 ttgctggggcgcggcgatgcggacgtgattttcgtcaccgacggccatgaggcgccgcca 480 ctcaatccccgctactgcccggacttcagcgacctcagaggcaaggtccgggggctgatc 540 gtcggagtgggaggactgagcctctcgcccatccccaagtacgacgagtcggggcggcgt 600 tcgggcgtttatggcgaggacgaagtcccgcagcgctcgagcttcggcctgtcggagctg 660 ccgcccgagcagatcgagggctaccacgcccgcaacgctcccttcggcagcgagagagcc 720 gggggcacggaacatctgtcccagctcaaggaaggatatttgcgccagctcgccgaagcc 780 gccggcctgggctaccaccgcctggaatcgcccgaaggactgggccgcgctctcacggca 840 ccggccttggcgcggcgccagcggatcgccacagacgtccgctggattcccgccgccctg 900 gcgctcgccgtactgatggcggtgtatctgcgggtgctgctgccgcgtcctggattttca 960 acctcaaactga 972 3-52 Sequences 3-52-1 SequenceNumber[ID] 52 3-52-2 MoleculeType AA 3-52-3 Length 323 source1...323 3-52-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-52-5 Residues LSIWRQRVADPVFAGLIVALLLAVAACFPLRLVLERLVFSHIVVVDITRSMNVEDYRRGA 60 RAVSRLEFVRQSLIGAVADLPCGSAVGVGVFTEREPALLFEPIETCAGFSAISAAIEQLD 120 WRMAWAADSLIAAGLHNTLDLLGRGDADVIFVTDGHEAPPLNPRYCPDFSDLRGKVRGLI 180 VGVGGLSLSPIPKYDESGRRSGVYGEDEVPQRSSFGLSELPPEQIEGYHARNAPFGSERA 240 GGTEHLSQLKEGYLRQLAEAAGLGYHRLESPEGLGRALTAPALARRQRIATDVRWIPAAL 300 ALAVLMAVYLRVLLPRPGFSTSN 323 3-53 Sequences 3-53-1 SequenceNumber[ID] 53 3-53-2 MoleculeType DNA 3-53-3 Length 525 source1...525 3-53-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-53-5 Residues atgaaaccgatgctcatcctgaccgcgttgctgttcgcctccgtttcgttggcgcacgga 60 cccaccccccaaaaggtcgtcgagaccgtggagatcgcggctcccgtggaccgggtctgg 120 aacgccgtgaaggatttcggtgccatcgcgcagtggaatcccgctctggccaagagcgaa 180 agcaccggcggcaacaccaccggcgagaagcgcatcctccattttcccaacggcgagcag 240 ctcaccgaggaactcgatgcctacgacccggcagcccacgaatacacctaccggctgggc 300 aaggacaacgtcaaggcgctgccggccagttcctactccgccgtgctcaaggtcaaggcc 360 accgagacgggcagccagatcgaatggaagagtcggctctatcgcggcgataccggaaac 420 ttcccgccggacgagctgaacgacgaggccgccgttgcggcgatgcagaggtttttccgc 480 gccgggctggacaatctcaagaaaagtcttgggcccctcgaatga 525 3-54 Sequences 3-54-1 SequenceNumber[ID] 54 3-54-2 MoleculeType AA 3-54-3 Length 174 source1...174 3-54-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-54-5 Residues MKPMLILTALLFASVSLAHGPTPQKVVETVEIAAPVDRVWNAVKDFGAIAQWNPALAKSE 60 STGGNTTGEKRILHFPNGEQLTEELDAYDPAAHEYTYRLGKDNVKALPASSYSAVLKVKA 120 TETGSQIEWKSRLYRGDTGNFPPDELNDEAAVAAMQRFFRAGLDNLKKSLGPLE 174 3-55 Sequences 3-55-1 SequenceNumber[ID] 55 3-55-2 MoleculeType DNA 3-55-3 Length 1530 source1...1530 3-55-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-55-5 Residues atgctgcaaaaatacatagagaagattctgcgcgcccgtgtctacgacgttgcccaggag 60 accccgctggacccggcgcccggcctgtcgcggcggctggacaacacggtgctgatcaag 120 cgcgaggacctgcagccggtgttctcgttcaagctgcgcggcgcctacaacaagatcgcc 180 tcgctcacacccgaggcgcgcgcggccggcgtgatcgcggcctccgccggcaaccacgcc 240 cagggcgtggcactggcggcgcagcggctgggcatccgcgccgtgatcgtgatgccttgc 300 accaccccgcatatcaaggtcgatgcggtgcgcaaccgaggcggtgaggtcgtactgcat 360 ggcgacgcctatgacgaagcctacgaacatgcgctggaactggcccgcgaccagtgcctg 420 accttcgtccacccctacgacgatccggaagtcatcgccgggcaaggcaccatcggcatg 480 gaaatcctgcgccagcaccaggacgccatccacgccatcttcgtgcctgtgggcggcggc 540 ggattgatcgccggcatcgccgcctacgtcaagttcgtgcgcccggacatccgcgtcatc 600 ggcgtggaaccagtggactccgactgcctgcaccgggcgctgaaagccaagcggcgggtg 660 atcctgaagcaggtgggcctgttcgccgacggcgtcgcggtgaagcaggtcggcaaggaa 720 ccgttccatctcgcccaccagtgggtggacgaggtcgtgaccgtcgacaccgacgaaatc 780 tgcgccgccatcaaggacatcttcgacgacacccgctccatcgccgagccggcgggcgcg 840 ctgggcatcgccgggctcaagaaatacgtggccgaaacaggaatcaagaacgcgtgcctg 900 gtggcgatcgaaagcggcgccaacatcaacttcgaccggctgcgccacgtcgctgagcgc 960 gccgagatcggcgaaaagcgcgaactgctgctggcagtgacgatccccgagcggcccggc 1020 agcttcctcgaattctgccgggtgctgggccgccgcaacatcaccgaattcaactaccgc 1080 ttcttcgacgaaaaggccgcccaggtgttcgtcggcctcccggtggcgagcggcgcgatc 1140 gaccgcgaaagcctggtccgcgaattcgaacgccagggtttcggcgtgctcgacctgacc 1200 ggcaacgaactcgccatcgaacacatccgctacatggtcggcggccacgcgccgaaactg 1260 ctggacgaacaggtctacagcttcgaattccccgagcgacccggcgcgctgctgcgcttc 1320 ctgtccatcatgggcgggcgctggaacatcagcctgttccattaccgcaaccacggcgcc 1380 gccttcggccgggtactgatgggcatccaggtgccgaaaccggaacgcaaggccttccgg 1440 gaattcctcgaagccatcggctacgccttcaaggaggaaacccaaaatcccgcctaccgg 1500 ctgttcgcggggggcagcgagcgggggtga 1530 3-56 Sequences 3-56-1 SequenceNumber[ID] 56 3-56-2 MoleculeType AA 3-56-3 Length 509 source1...509 3-56-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-56-5 Residues MLQKYIEKILRARVYDVAQETPLDPAPGLSRRLDNTVLIKREDLQPVFSFKLRGAYNKIA 60 SLTPEARAAGVIAASAGNHAQGVALAAQRLGIRAVIVMPCTTPHIKVDAVRNRGGEVVLH 120 GDAYDEAYEHALELARDQCLTFVHPYDDPEVIAGQGTIGMEILRQHQDAIHAIFVPVGGG 180 GLIAGIAAYVKFVRPDIRVIGVEPVDSDCLHRALKAKRRVILKQVGLFADGVAVKQVGKE 240 PFHLAHQWVDEVVTVDTDEICAAIKDIFDDTRSIAEPAGALGIAGLKKYVAETGIKNACL 300 VAIESGANINFDRLRHVAERAEIGEKRELLLAVTIPERPGSFLEFCRVLGRRNITEFNYR 360 FFDEKAAQVFVGLPVASGAIDRESLVREFERQGFGVLDLTGNELAIEHIRYMVGGHAPKL 420 LDEQVYSFEFPERPGALLRFLSIMGGRWNISLFHYRNHGAAFGRVLMGIQVPKPERKAFR 480 EFLEAIGYAFKEETQNPAYRLFAGGSERG 509 3-57 Sequences 3-57-1 SequenceNumber[ID] 57 3-57-2 MoleculeType DNA 3-57-3 Length 1545 source1...545 3-57-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-57-5 Residues atgcacgacagactgatcattttcgacacgaccttgcgcgacggagagcagagccccggc 60 gcgtccatgacccgcgatgaaaaggtccgcatcgcccgggcgctggagcgtctgaaggtc 120 gacgtcatcgaggcgggctttcccgccgccagccccggcgatttcgaggccgtccaggcc 180 gtggcccggaccatcaaggacagcagggtctgcggcctggcccgcgccctcgaccgcgac 240 atcgaccgcgccggcgaagccctcaaggacgcccagcgcgcccgcatccacaccttcatc 300 gccacctcgcccatccacatgcggcacaagctgcagatgtcgcccgaccaggtggtggaa 360 tacgcggtcaaggccgtcaagcgggcccgccagtacaccgacgacgtggaattctcgccc 420 gaggacgccggacgctccgaggaggatttcctctgccgcatcctggaagccgtgatcgat 480 gcgggggcgaccacgctgaacatccccgacaccgtcggctacgccttcccggaacagttc 540 gggcacatgatcggccggctgatcgagcggattccgaactccgacaaggccgtgttctcg 600 gttcactgccacaacgacctgggactggcggtcgccaattcgctggccgccgtgctgcac 660 ggcgcgcgccaggtggaatgcaccatcaacgggctgggcgagcgggccggcaacgccgcg 720 ctggaagagatcgtcatggcggtgcgcacccgtaaagacatcttcccctgccacaccgac 780 atcgagacacgggaaatcgtggcctgctccaaactggtctccagcatcaccggtttcccg 840 atccagcccaacaaggccatcgtcggcgccaacgccttcgcccacgagtcgggcatccac 900 caggacggtgtgctcaagagccgggaaacctacgagatcatgagcgccgaggacgtgggg 960 tggagcaccaaccgcatggtgctgggcaaacattccggccgcaacgcgttccgtacccgg 1020 atgcaggaactcggcatcgagttcgcctcggaagaggaactgaactcggtgttccagcgc 1080 ttcaaggtgctggccgacaagaagcacgagatcttcgacgaggacctccaggccctcatc 1140 accgaagccggcgcagaagccgaagacgaacgggtcaagctggtcgcgctgcgggtctgc 1200 tcggaaacgggcgagattccccacgcccaggtcaccatcaaggtggacaacgaggaacgc 1260 accggcacatcgagcggcggcggcgccgtggacgccagcctcaaggccatcgaatcgctg 1320 ctgcacacggacaccgcgctgacgctgtactcggtcaacaacatcaccagcggcaccgac 1380 gcccagggcgaggtcaccgtgcggctcgagaaaggcgggcgcatcgtcaacggccagggc 1440 gccgataccgacatcgtgatcgcctcggccaaggcctacgtcaacgccgtgaacaagctg 1500 ctggcgcccatccagcgcacccacccgcaagtcggggatgtgtga 1545 3-58 Sequences 3-58-1 SequencesNumber[ID] 58 3-58-2 MoleculeType AA 3-58-3 Length 514 source1...514 3-58-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-58-5 Residues MHDRLIIFDTTLRDGEQSPGASMTRDEKVRIARALERLKVDVIEAGFPAASPGDFEAVQA 60 VARTIKDSRVCGLARALDRDIDRAGEALKDAQRARIHTFIATSPIHMRHKLQMSPDQVVE 120 YAVKAVKRARQYTDDVEFSPEDAGRSEEDFLCRILEAVIDAGATTLNIPDTVGYAFPEQF 180 GHMIGRLIERIPNSDKAVFSVHCHNDLGLAVANSLAAVLHGARQVECTINGLGERAGNAA 240 LEEIVMAVRTRKDIFPCHTDIETREIVACSKLVSSITGFPIQPNKAIVGANAFAHESGIH 300 QDGVLKSRETYEIMSAEDVGWSTNRMVLGKHSGRNAFRTRMQELGIEFASEEELNSVFQR 360 FKVLADKKHEIFDEDLQALITEAGAEAEDERVKLVALRVCSETGEIPHAQVTIKVDNEER 420 TGTSSGGGAVDASLKAIESLLHTDTALTLYSVNNITSGTDAQGEVTVRLEKGGRIVNGQG 480 ADTDIVIASAKAYVNAVNKLLAPIQRTHPQVGDV 514 3-59 Sequences 3-59-1 SequencesNumber[ID] 59 3-59-2 MoleculeType DNA 3-59-3 Length 1413 source1...413 3-59-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-59-5 Residues atgagcggaaaaaccctttacgacaagctgtgggacgaccacgtcgtgcatgtcgatgcg 60 gacggatcgtgcctgatctacatcgatcgtcatctaatccacgaggtgacctcgcctcag 120 gcattcgaagggctgcggatggcggggcgtgtaccctggcgggtggatgccaatcttgcg 180 gtggccgaccacaacgtccccaccgccgaccgcgacaggggtatcgccgatccggtgtcg 240 cgcctgcaggtggaaaccctggacaagaactgcgccgatttcggcatcaccgaattcgcg 300 atggacgacgtgcgccagggtatcgtgcatgtgatcgggcccgagcagggcgcgaccctg 360 ccgggcatgaccatcgtttgcggcgattcgcatacttcgactcacggtgctttcggggcg 420 ctcgccttcgggatcggcacttccgaggtcgagcacgtactggccacgcaatgcctggtg 480 cagcgcaaggcgaagaacatgctggtccgcgtccagggcaagctggcgccgggcgtgacg 540 gcgaaagatctggtactggcggtcatcggccgtatcggaaccgccggcggcaccggctac 600 accatcgaattcgctggcgaagccattcgcggcctgtcgatggaaggccggatgacggtc 660 tgcaacatggcgatcgaggcgggcgcacgtgccggcctggtggcggtggacgaagtcacg 720 ctcgactatctcgagggccgcccgttcgctccggcgggcgcgttgtgggagcgggcggtc 780 gaggcatggaaagacctgcacagcgatccggatgcggtattcgacaaggtcgtcgagatc 840 gatgccgccagcatcaagccgcaggtgacctggggaacttcgccggaacaggtcgtgccg 900 gtggatgccgaggtgcccgacccggccacggaagccgatcccgtgcggcgggaaagcatg 960 gagcgggcgctgcagtacatggatctcctgccgggcacgccaatcggcgcgatccgggtc 1020 gatcgggtgttcatcggctcctgcaccaatgccaggatcgaggatctgcgcgccgcggcg 1080 gaagtcgtccgggggcacaagcgcgctgccagcgtgaagcaggcactggtggtgcccggc 1140 tcgggtttggtcaagcggcaggcggagcaggaggggctggacaaggtgttcctcgaggcc 1200 ggtttcgaatggcgcgacccgggttgttccatgtgtctggcgatgaacgccgaccgcctg 1260 gaacccggcgagcgttgcgcctcgacctccaaccggaattttgaggggcgccagggctat 1320 ggcgggcgtacccatctggtgagtccggccatggcggctgcggcggccattcacgggcat 1380 ttcgtcgacatcaccgaaggagggcgcgcatga 1413 3-60 Sequences 3-60-1 SequencesNumber[ID] 60 3-60-2 MoleculeType AA 3-60-3 Length 470 source1...470 3-60-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-60-5 Residues MSGKTLYDKLWDDHVVHVDADGSCLIYIDRHLIHEVTSPQAFEGLRMAGRVPWRVDANLA 60 VADHNVPTADRDRGIADPVSRLQVETLDKNCADFGITEFAMDDVRQGIVHVIGPEGGATL 120 PGMTIVCGDSHTSTHGAFGALAFGIGTSEVEHVLATQCLVQRKAKNMLVRVQGKIAPGVT 180 AKDLVLAVIGRIGTAGGTGYTIEFAGEAIRGLSMEGRMTVCNMAIEAGARAGLVAVDEVT 240 LDYLEGRPFAPAGALWERAVEAWKDLHSDPDAVFDKVVEIDAASIKPQVTWGTSPEQVVP 300 VDAEVPDPATEADPVRRESMERALQYMDLLPGTPIGAIRVDRVFIGSCTNARIEDLRAAA 360 EVVRGHKRAASVKQALVVPGSGLVKRQAEQEGLDKVFLEAGFEWRDPGCSMCLAMNADRL 420 EPGERCASTSNRNFEGRQGYGGRTHLVSPAMAAAAAIHGHFVDIEGGRA 470 3-61 Sequences 3-61-1 SequencesNumber[ID] 61 3-61-2 MoleculeType DNA 3-61-3 Length 639 source1...639 3-61-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-61-5 Residues atgaagcctttcaagaaattcacttcgcgagtcgtgccgttggaccgcgccaatgtcgac 60 accgacgccatcattcccaagcagttcctgaagtccatccgccgcagcgggttcggtccc 120 tatctgttcgacgagtggcgttacctggaccgtggcgagcccgacatggattgcagccac 180 cgtccgctcaacccggagttcgtgctcaacctgccctgttacgccggcgccaggatattg 240 ctggcccgcaagaacttcggctgtggctcctcgcgcgagcatgcgccctgggcgctggag 300 gattacggcttccgcgccatcatcgcgccgagtttcgccgatatcttctacaacaactgc 360 ttcaagaacggcatcctgcccatcgtgctcgacgaggccacggtcgaccggctgtttagc 420 gaggccgggcccggcttcgagctcaccgtcgacctggagtcgcagaccgtggcgacgccg 480 ttcggcgagaccttccatttcgacgtggatgcctcccgcaagcatcgtctgctgaacggc 540 ctggacgacatcggtctgacccttcagcatgccgatgccatccgcgcctacgaagccgcc 600 cgcaggaagtccgcaccctggctgtttgccgtcccttga 639 3-62 Sequences 3-62-1 SequencesNumber[ID] 62 3-62-2 MoleculeType AA 3-62-3 Length 212 source1...212 3-62-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-62-5 Residues MKPFKKFTSRVVPLDRANVDTDAIIPKQFLKSIRRSGFGPYLFDEWRYLDRGEPDMDCSH 60 RPLNPEFVLNLPCYAGARILLARKNFGCGSSREHAPWALEDYGFRAIIAPSFADIFYNNC 120 FKNGILPIVLDEATVDRLFSEAGPGFELTVDLESQTVATPFGETFHFDVDASRKHRLLNG 180 LDDIGLTLQHADAIRAYEAARRKSAPWLFAVP 212 3-63 Sequences 3-63-1 SequenceNumber[ID] 63 3-63-2 MoleculeType DNA 3-63-3 Length 1083 source1...1083 3-63-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-63-5 Residues atgactatcaaaatcgctgtcttgcccggtgacggcatcggtcccgaaatcgtcgccgag 60 gccctgaaggttctggactgcctgcggtccgacttcggcctcgcggtcgaaaccgaacac 120 gccctgatcggcggcgcagcctatgatgcgcacggcacgccgttccccaaggaaaccctg 180 gagctgtgccgggctgccgattcgatcctgcttggagcggtcggcggtcccaaatgggag 240 ccgttggattattcgctgcggcccgagcgggggctcctgggcttgcgttcggagctggaa 300 ctgttttccaacctgcgcccggcggtgctctaccctcagctggtgtcggcttcgaccctc 360 aagcccgaggtggtcgccggcctcgacatcatgatcgtgcgggagctgaccggcggcata 420 tatttcggcaagccgcgcggtcgtcgcatcaacgaggacggagagcgggagggctacaac 480 accctggtatacagcgaatcggaaatccgccgcatagcccatagcgcgttccagatcgcc 540 cggaagcgtaacaggcgcctgtgcagcatcgacaaggccaatgtgctggaatgcacggaa 600 ctgtggcgcgaggtggtgatcgaggtcggcaaggactatcccgacgtggcgctgagccac 660 atgtacgtggacaacgccgcgatgcagctggtccgtaacccgaagcagttcgacgtgatg 720 ctgaccgacaacatgttcggcgacatcctgtccgactgtgccgccatgctgaccggctcg 780 atcggcatgctgccttcggcttccctcgccgagagcggcaaggggatgtacgagcccatc 840 cacggttcggccccggatatcgccggccgcggcatcgccaacccgatcgccaccatcctg 900 tcgctggccatgatgttgcgctacagcttcgatgacgcggtctcggcagagcggatcggg 960 aaggcggtgcagacggcgctggatcagggtttccgcacggcggacatcgcctcggaaggc 1020 accgtcgaggtcggtaccgctgcgatgggcgatgccatcgtcgccgccttgcgcgccgtc 1080 tga 1083 3-64 Sequences 3-64-1 SequenceNumber[ID] 64 3-64-2 MoleculeType AA 3-64-3 Length 360 source1...360 3-64-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-64-5 Residues MTIKIAVLPGDGIGPEIVAEALKVLDCLRSDFGLAVETEHALIGGAAYDAHGTPFPKETL 60 ELCRAADSILLGAVGGPKWEPLDYSLRPERGLLGLRSELELFSNLRPAVLYPQLVSASTL 120 KPEVVAGLDIMIVRELTGGIYFGKPRGRRINEDGEREGYNTLVYSESEIRRIAHSAFQIA 180 RKRNRRLCSIDKANVLECTELWREVVIEVGKDYPDVALSHMYVDNAAMQLVRNPKQFDVM 240 LTDNMFGDILSDCAAMLTGSIGMLPSASLAESGKGMYEPIHGSAPDIAGRGIANPIATIL 300 SLAMMLRYSFDDAVSAERIGKAVQTALDQGFRTADIASEGTVEVGTAAMGDAIVAALRAV 360 3-65 Sequences 3-65-1 SequenceNumber[ID] 65 3-65-2 MoleculeType DNA 3-65-3 Length 1477 source1...1477 3-65-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Marinobacteralgicola NonEnglishQualifierValue 3-65-5 Residues atggcgacccagcagcagcagaacggcgcctcggcgagcggcgtcctggaacagttgcgc 60 gggaagcatgtcctgataaccggtaccaccggtttccttggcaaggtagtcctggaaaag 120 ctgatccgcacagtcccggacatcggcggcatccacctcctgatccggggcaacaagagg 180 catccggccgcccgtgaacggttcttgaacgagatcgccagcagttcggtcttcgagcgt 240 ctgcgccacgacgacaacgaggccttcgaaaccttcctggaagaaagggtgcactgtata 300 accggagaggtcaccgagagtcgtttcggccttaccccggagcgcttccgcgcgctggcg 360 ggtcaggtggacgccttcatcaattcggccgcctccgtcaacttccgcgaggaactggac 420 aaggcgctgaagatcaatacgctgtgcctggagaatgtcgcggcccttgctgaactcaac 480 agtgcgatggcggtcatccaggtttcgacctgctacgttaacggcaagaatagcgggcag 540 atcaccgaatcggtcatcaagcccgcgggggagtccatcccgcgtagcaccgatgggtac 600 tatgaaatcgaagaattggtgcacctgctgcaggacaaaatcagcgatgtgaaggcccga 660 tactccgggaaggttctggaaaaaaaattggtggacctaggcatccgggaagccaataac 720 tacgggtggagcgatacatataccttcaccaagtggctgggcgaacagctcctcatgaag 780 gccctgagcggcagatcgctgaccatcgtgcggccgtcgatcatcgagtcggcattggaa 840 gagcccagcccggggtggattgaaggcgtcaaggtcgccgatgccatcatactggcctac 900 gcgagggagaaggtatcgctctttcctggcaagcggagcggcatcatcgacgtcatccca 960 gtggatctggtggccaattcgatcattctgtccctggcggaggcgctctccggttcgggc 1020 cagcggcgtatctatcagtgctgcagcggcggctcgaaccccatctccctcgggaagttc 1080 atcgactatctgatggcggaggcgaagaccaactacgcggcctacgatcagctgttctac 1140 cgccgccccaccaagccgttcgtggccgtcaaccgcaaactcttcgacgtcgtcgtgggc 1200 ggcatgcgggtcccgctctcgatcgcgggcaaagccatgcgcctggcgggacaaaaccgc 1260 gaactgaaggtcctgaagaatctggatacgacccggtccctggccaccattttcgggttc 1320 tacaccgctccggactacatctttcgcaatgacagcctgatggccctggcctcgcgcatg 1380 ggcgagctggaccgcgtgttgttccccgttgacgcccgtcagatcgactggcagctgtat 1440 ctgtgcaaaatccacctcggcgggctgaatcggtacg 1477 3-66 Sequences 3-66-1 SequenceNumber[ID] 66 3-66-2 MoleculeType AA 3-66-3 Length 55 source1...511 3-66-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Marinobacteralgicola NonEnglishQualifierValue 3-66-5 Residues MATQQQQNGASASGVLEQLRGKHVLITGTTGFLGKVVLEKLIRTVPDIGGIHLLIRGNKR 60 HPAARERFLNEIASSSVFERLRHDDNEAFETFLEERVHCITGEVTESRFGLTPERFRALA 120 GQVDAFINSAASVNFREELDKALKINTLCLENVAALAELNSAMAVIQVSTCYVNGKNSGQ 180 ITESVIKPAGESIPRSTDGYYEIEELVHLLQDKISDVKARYSGKVLEKKLVDLGIREANN 240 YGWSDTYTFTKWLGEQLLMKALSGRSLTIVRPSIIESALEEPSPGWIEGVKVADAIILAY 300 AREKVSLFPGKRSGIIDVIPVDLVANSIILSLAEALSGSGQRRIYQCCSGGSNPISLGKF 360 IDYLMAEAKTNYAAYDQLFYRRPTKPFVAVNRKLFDVVVGGMRVPLSIAGKAMRLAGQNR 420 ELKVLKNLDTTRSLATIFGFYTAPDYIFRNDSLMALASRMGELDRVLFPVDARQIDWQLY 480 LCKIHLGGLNRYALKERKLYSLRAADTRKKA 511 3-67 Sequences 3-67-1 SequenceNumber[ID] 67 3-67-2 MoleculeType DNA 3-67-3 Length 1377 source1...1377 3-67-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Acinetobacterbaylyi NonEnglishQualifierValue 3-67-5 Residues atgcgccccctgcaccccatcgacttcatcttcctgagcctggaaaagcggcagcagccc 60 atgcacgtcggcggcctgttcctgttccagatcccggacaacgcccccgacaccttcatc 120 caggacctggtcaacgacatccgcatctccaagagcatcccggtgccgcccttcaacaac 180 aagctgaacggcctgttctgggacgaagacgaggagttcgacctggaccaccatttccgg 240 cacatcgccctgccgcatcccggccgcatccgggaactgctgatctacatctcccaggag 300 cacagcaccctgctggaccgcgcgaagccgctgtggacctgcaacatcatcgaaggcatc 360 gagggcaaccggttcgccatgtatttcaagatccaccatgcgatggtcgacggcgtggcc 420 ggcatgcgcctgatcgaaaagtcgctgtcccatgacgtcaccgagaagagcatcgtcccg 480 ccctggtgcgtggaaggcaagcgggcgaagcgcctgcgggagccgaagaccggcaagatc 540 aagaagatcatgtcgggcatcaagtcccagctgcaggccacccccaccgtcatccaggaa 600 ctgtcgcagaccgtgttcaaggacatcggccgcaacccggaccacgtcagctcgttccag 660 gccccctgctccatcctgaaccagcgggtgtccagctcgcgccggttcgccgcgcagtcg 720 ttcgacctggaccgcttccggaacatcgcgaagtccctgaacgtcaccatcaacgacgtc 780 gtgctggccgtgtgcagcggcgccctgcgcgcgtacctgatgagccacaactcgctgccg 840 tccaagcccctgatcgcgatggtcccggcgtcgatccgcaacgacgacagcgacgtgtcg 900 aaccggatcaccatgatcctggccaacctggcgacccataaggacgacccgctgcagcgc 960 ctggagatcatccgccggagcgtccagaactcgaagcagcgcttcaagcggatgacctcc 1020 gaccagatcctgaactacagcgcggtcgtgtatggcccggccggcctgaacatcatcagc 1080 ggcatgatgcccaagcgccaggccttcaacctggtcatctcgaacgtgccgggcccgcgc 1140 gagccgctgtactggaacggcgccaagctggacgcgctgtatcccgcctccatcgtcctg 1200 gacggccaggccctgaacatcaccatgaccagctacctggacaagctggaggtcggcctg 1260 atcgcgtgccgcaacgccctgccgcggatgcagaacctgctgacccatctggaggaagag 1320 atccagctgttcgaaggcgtgatcgcgaagcaggaggacatcaagaccgccaactga 1377 3-68 Sequences 3-68-1 SequenceNumber[ID] 68 3-68-2 MoleculeType AA 3-68-3 Length 458 source1...458 3-68-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Acinetobacterbaylyi NonEnglishQualifierValue NonEnglishQualifierValue NonEnglishQualifierValue 3-68-5 Residues MRPLHPIDFIFLSLEKRQQPMHVGGLFLFQIPDNAPDTFIQDLVNDIRISKSIPVPPFNN 60 KLNGLFWDEDEEFDLDHHFRHIALPHPGRIRELLIYISQEHSTLLDRAKPLWTCNIIEGI 120 EGNRFAMYFKIHHAMVDGVAGMRLIEKSLSHDVTEKSIVPPWCVEGKRAKRLREPKTGKI 180 KKIMSGIKSQLQATPTVIQELSQTVEKDIGRNPDHVSSFQAPCSILNQRVSSSRRFAAQS 240 FDLDRFRNIAKSLNVTINDVVLAVCSGALRAYLMSHNSLPSKPLIAMVPASIRNDDSDVS 300 NRITMILANLATHKDDPLQRLEIIRRSVQNSKQRFKRMTSDQILNYSAVVYGPAGLNIIS 360 GMMPKRQAFNLVISNVPGPREPLYWNGAKLDALYPASIVLDGQALNITMTSYLDKLEVGL 420 IACRNALPRMQNLLTHLEEEIQLFEGVIAKQEDIKTAN 458 3-69 Sequences 3-69-1 SequenceNumber[ID] 69 3-69-2 MoleculeType DNA 3-69-3 Length 1428 source1...1428 3-69-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Psychrobacterarticus NonEnglishQualifierValue 3-69-5 Residues atgcgcctgctgaccgccgtcgaccagctgttcctgctgctggagtcccgcaagcacccg 60 atgcacgtgggcggcctgttcctgttcgaactgccggagaacgccgacatctcgttcgtc 120 caccagctggtgaagcagatgcaggactccgacgtcccgcccaccttccccttcaaccag 180 gtgctggaacacatgatgttctggaaggaggacaagaacttcgacgtcgaacaccatctg 240 caccatgtggccctgccgaagcccgcgcgcgtccgggagctgctgatgtacgtgtcccgc 300 gaacacggccggctgctggaccgcgcgatgccgctgtgggaatgccatgtcatcgagggc 360 atccagccggaaaccgagggcagccccgagcggttcgccctgtatttcaagatccaccat 420 tcgctggtcgacggcatcgccgcgatgcgcctggtgaagaagagcctgtcgcagtcgccg 480 aacgaacccgtgaccctgccgatctggagcctgatggcccaccatcggaaccagatcgac 540 gcgatcttccccaaggagcggagcgccctgcgcatcctgaaggaacaggtctcgaccatc 600 aagccggtgttcaccgagctgctgaacaacttcaagaactacaacgacgactcgtatgtc 660 tccaccttcgacgcgccccgcagcatcctgaaccgccggatcagcgcctcgcgccggatc 720 gccgcgcagtcgtacgacatcaagcggttcaacgacatcgccgaacgcatcaacatctcc 780 aagaacgacgtcgtgctggccgtgtgcagcggcgcgatccgccgctacctgatcagcatg 840 gacgcgctgccgagcaagcccctgatcgccttcgtcccgatgtcgctgcgcaccgacgac 900 tccatcgcgggcaaccagctgtcgttcgtgctggccaacctgggcacccacctggacgac 960 cccctgtcccggatcaagctgatccatcgctccatgaacaacagcaagcgccggttccgc 1020 cggatgaaccaggcccaggtcatcaactacagcatcgtgtcgtatgcctgggagggcatc 1080 aacctggcgaccgacctgttcccgaagaagcaggccttcaacctgatcatctcgaacgtg 1140 ccgggcagcgagaagcccctgtactggaacggcgcgcgcctggaaagcctgtatccggcc 1200 tcgatcgtgttcaacggccaggccatgaacatcaccctggcgtcctacctggacaagatg 1260 gagttcggcatcaccgcctgcagcaaggcgctgccgcacgtccaggacatgctgatgctg 1320 atcgaggaagagctgcagctgctggagtccgtcagcaaggaactggagttcaacggcatc 1380 accgtgaaggacaagtcggaaaagaagctgaagaagctggccccgtga 1428 3-70 Sequences 3-70-1 SequenceNumber[ID] 70 3-70-2 MoleculeType AA 3-70-3 Length 475 source1...475 3-70-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Psychrobacterarticus NonEnglishQualifierValue 3-70-5 Residues MRLLTAVDQLFLLLESRKHPMHVGGLFLFELPENADISFVHQLVKQMQDSDVPPTFPFNQ 60 VLEHMMFWKEDKNFDVEHHLHHVALPKPARVRELLMYVSREHGRLLDRAMPLWECHVIEG 120 IQPETEGSPERFALYFKIHHSLVDGIAAMRLVKKSLSQSPNEPVTLPIWSLMAHHRNQID 180 AIFPKERSALRILKEQVSTIKPVFTELLNNFKNYNDDSYVSTFDAPRSILNRRISASRRI 240 AAQSYDIKRFNDIAERINISKNDVVLAVCSGAIRRYLISMDALPSKPLIAFVPMSLRTDD 300 SIAGNQLSFVLANLGTHLDDPLSRIKLIHRSMNNSKRRFRRMNQAQVINYSIVSYAWEGI 360 NLATDLFPKKQAFNLIISNVPGSEKPLYWNGARLESLYPASIVFNGQAMNITLASYLDKM 420 EFGITACSKALPHVQDMLMLIEEELQLLESVSKELEFNGITVKDKSEKKLKKLAP 475 3-71 Sequences 3-71-1 SequenceNumber[ID] 71 3-71-2 MoleculeType DNA 3-71-3 Length 1386 source1...1386 3-71-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Rhodococcusopacus NonEnglishQualifierValue 3-71-5 Residues atgtccgtgatgtccccgaccgaggcgatgttcgtcctgttcgagaccccgagccacccg 60 atgcacatgggcgcgctggagctgttcgagccgccgcgcgagtcgggcccggaccacgcc 120 cgcctgatgttcgaggcgctgatctcccaggaaggcgccagcgacaccttccgccggcgc 180 gccgtccggccgctgcgcggcgcgtcgtacccctggtggtccgtcgacgaccgggtggac 240 ctgggctatcacgtccgccataccgccgtgccgggccggggccgcatggaggacctgctg 300 tcgctggtgtcccagatgcacggcatgcccctggacccgcagcaccccatgtgggagatc 360 catgtcatcgaaggcctggccgacggccgcaccgcggtgttcagcaagatccatctgtcg 420 ctgatggacggcccggccggcctgcggctgctgcaccatgcgctgagcaccgacccggac 480 gcccgcgactgccccgcgccgtggacccccggcgtcagcggcacctcgcggcgcgaatcg 540 gccctgccggtcgccgcggtgcgggcgggcgtgcgcgccgcgacctccatcgtcggcgtg 600 ctgcccgccctggcgaaggtcgcctacgacggcgtgcgggaccagcacctgaccctgccg 660 ctgcagagcccgcccaccatgctgaacgtccccgtgggccgggcccgcaagctggccgcg 720 cggagctggccgatccggcgcctggtctcggtggccgcggccgcgcgcaccaccatcaac 780 gccgtcgtgctggcgatgtgctcgggcgccctgcgccactacctggtcgagcagtatgcc 840 ctgccggaagcgcccctgaccgccatgctgcccgtgccgctggacctgggcggcaccatg 900 atcggcccgcgtggccgcgaccacggcgtcggcgcgatggtcgtgggcctggcgaccgac 960 gaggccgaccccgccgcgcggctggcccgcatcagcgagtcggtcgaacacaccaaccgc 1020 gtgttcggcgcgctgtcccatacccagttccaggtcatgtccgccctggcgatcagcccg 1080 atcctgctggaacccgtccggcgcttcgtggacgacaccccgcccccgttcaacgtgatg 1140 atctcgtacatgccgggtccgtcccggccgcgctattggaacggcgcgcggctggacgcc 1200 gtctaccccgcgccgaccgtgctgggcggccaggccctgagcatcaccctgacctcccgc 1260 agcggccagctggacgtcggcgtcgtgggcgaccggcaggccgtgccgcacctgcagcgc 1320 atcatcacccatctggagacctccctgaccgacctggaaaacgccgtggccgcgagcggc 1380 acctga 1386 3-72 Sequences 3-72-1 SequenceNumber[ID] 72 3-72-2 MoleculeType AA 3-72-3 Length 461 source1...461 3-72-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Rhodococcusopacus NonEnglishQualifierValue 3-72-5 Residues MSVMSPTEAMFVLFETPSHPMHMGALELFEPPRESGPDHARLMFEALISQEGASDTFRRR 60 AVRPLRGASYPWWSVDDRVDLGYHVRHTAVPGRGRMEDLLSLVSQMHGMPLDPQHPMWEI 120 HVIEGLADGRTAVFSKIHLSLMDGPAGLRLLHHALSTDPDARDCPAPWTPGVSGTSRRES 180 ALPVAAVRAGVRAATSIVGVLPALAKVAYDGVRDQHLTLPLQSPPTMLNVPVGRARKLAA 240 RSWPIRRLVSVAAAARTTINAVVLAMCSGALRHYLVEQYALPEAPLTAMLPVPLDLGGTM 300 IGPRGRDHGVGAMVVGLATDEADPAARLARISESVEHTNRVFGALSHTQFQVMSALAISP 360 ILLEPVRRFVDDTPPPFNVMISYMPGPSRPRYWNGARLDAVYPAPTVLGGQALSITLTSR 420 SGQLDVGVVGDRQAVPHLQRIITHLETSLTDLENAVAASGT 461 3-73 Sequences 3-73-1 SequenceNumber[ID] 73 3-73-2 MoleculeType DNA 3-73-3 Length 1356 source1...1356 3-73-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Rhodococcusopacus NonEnglishQualifierValue 3-73-5 Residues atgccggtcaccgactccatcttcctgctgggcgaaagccgcgagcacccgatgcacgtg 60 ggctccctggaactgttcacccccccggacgacgccggcccggactacgtcaagtcgatg 120 cacgagaccctgctgaagcataccgacgtggaccccaccttccgcaagaagccggcgggc 180 cccgtcggctcgctgggcaacgtgtggtgggccgacgagtccgacgtcgacctggaatac 240 cacgtgcgccatagcgcgctgccggccccctatcgcgtccgggaactgctgaccctgacc 300 tcgcggctgcacggcaccctgctggaccgccatcggccgctgtgggagatgtacctgatc 360 gaaggcctgagcgacggccgcttcgccatctataccaagctgcaccatagcctgatggac 420 ggcgtctcgggcctgcgcctgctgatgcggaccctgtcgaccgacccggacgtgcgcgac 480 gccccgcccccgtggaacctgccgcggcccgccgcggccaacggcgcggccccggacctg 540 tggtcggtcgtgaacggcgtccgccggaccgtcggcgacgtggccggcctggcgcccgcc 600 tccctgcgcatcgcgcggaccgcgatgggccagcacgacatgcgcttcccgtacgaggcg 660 ccccggaccatgctgaacgtgccgatcggcggcgcccgccggttcgcggcccagtcctgg 720 cccctggaacgcgtccatgccgtgcggaaggcggccggcgtcagcgtgaacgacgtcgtg 780 atggccatgtgcgcgggcgccctgcgcggctatctggaggaacagaacgcgctgccggac 840 gagcccctgatcgcgatggtcccggtgtccctgcgggacgaacagcaggcggacgccggc 900 ggcaacgccgtcggcgtgaccctgtgcaacctggcgaccgacgtcgacgaccccgccgag 960 cgcctgaccgcgatcagcgcctcgatgtcccagggcaaggaactgttcggcagcctgacc 1020 tcgatgcaggcgctggcctggtcggcggtgaacatgtccccgatcgccctgaccccggtc 1080 cccggcttcgtgcggttcacccccccgcccttcaacgtcatcatcagcaacgtgccgggc 1140 ccccgcaagaccatgtactggaacggctcccggctggacggcatctatccgaccagcgtc 1200 gtgctggacggccaggccctgaacatcaccctgaccaccaacggcggcaacctggacttc 1260 ggcgtcatcggctgccgccggtccgtgccgagcctgcagcgcatcctgttctacctggaa 1320 gcggccctgggcgagctggaagcggccctgctgtga 1356 3-74 Sequences 3-74-1 SequenceNumber[ID] 74 3-74-2 MoleculeType AA 3-74-3 Length 451 source1...451 3-74-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Rhodococcusopacus NonEnglishQualifierValue 3-74-5 Residues MPVTDSIFLLGESREHPMHVGSLELFTPPDDAGPDYVKSMHETLLKHTDVDPTFRKKPAG 60 PVGSLGNVWWADESDVDLEYHVRHSALPAPYRVRELLTLTSRLHGTLLDRHRPLWEMYLI 120 EGLSDGRFAIYTKLHHSLMDGVSGLRLLMRTLSTDPDVRDAPPPWNLPRPAAANGAAPDL 180 WSVVNGVRRTVGDVAGLAPASLRIARTAMGQHDMRFPYEAPRTMLNVPIGGARRFAAQSW 240 PLERVHAVRKAAGVSVNDVVMAMCAGALRGYLEEQNALPDEPLIAMVPVSLRDEQQADAG 300 GNAVGVTLCNLATDVDDPAERLTAISASMSQGKELFGSLTSMQALAWSAVNMSPIALTPV 360 PGFVRFTPPPFNVIISNVPGPRKTMYWNGSRLDGIYPTSVVLDGQALNITLTTNGGNLDF 420 GVIGCRRSVPSLQRILFYLEAALGELEAALL 451 3-75 Sequences 75 3-75-1 SequenceNumber[ID] DNA 3-75-2 MoleculeType 1395 3-75-3 Length source1..1395 3-75-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Rhodococcusopacus NonEnglishQualifierValue 3-75-5 Residues atgcccctgccgatgtcccccctggactccatgttcctgctgggcgaaagccgcgagcac 60 ccgatgcacgtgggcggcgtcgaaatcttccagctgcccgagggcgccgacacctacgac 120 atgcgggcgatgctggaccgcgccctggcggacggcgacggcatcgtcaccccgcggctg 180 gccaagcgcgcgcgccggtcgttcagctcgctgggccagtggtcctgggaaaccgtggac 240 gacatcgacctgggccaccatatccggcacgacgccctgccggcccctggcggcgaggcc 300 gaactgatggcgctgtgctcgcgcctgcacggctccctgctggaccgcagccggccgctg 360 tgggagatgcatctgatcgaaggcctgagcgacggccgcttcgccgtctataccaagatc 420 caccatgccgtcgcggacggcgtgaccgccatgaagatgctgcggaacgcgctgagcgag 480 aactcggacgaccgcgacgtgccggccccctggcagccgcgtggcccgcggccccagcgc 540 accccctccagcaagggcttctccctgagcggcctggccggctcgaccctgcggaccgcg 600 cgcgagaccgtcggcgaagtggccggcctggtcccggccctggcgggcaccgtgagccgg 660 gccttccgcgaccagggcggcccgctggccctgtcggcgccgaagacccccttcaacgtc 720 cccatcaccggcgcccgccagttcgccgcgcagtcgtggccgctggaacgcctgcggctg 780 gtggccaagctgtcggactccaccatcaacgacgtcgtgctggccatgtcgtccggcgcg 840 ctgcggtcctacctggaggaccagaacgccctgccggcggaccccctgatcgcgatggtc 900 ccggtgtccctgaagagccagcgcgaagccgcgaccggcaacaacatcggcgtcctgatg 960 tgcaacctgggcacccacctgcgggagccggccgaccgcctggaaaccatccggaccagc 1020 atgcgcgagggcaaggaagcctatggctcgatgaccgcgacccagatcctggccatgtcc 1080 gcgctgggcgccgcgccgatcggcgccagcatgctgttcggccataactcgcgcgtccgg 1140 ccgcccttcaacctgatcatctccaacgtgccgggccccagctcgccgctgtactggaac 1200 ggcgcccgcctggacgcgatctatccgctgagcgtccccgtggacggccagggcctgaac 1260 atcacctgcacctcgaacgacgacatcatctccttcggcgtcaccggctgccggtccgcc 1320 gtgccggacctgaagagcatccccgcgcgcctgggccatgagctgcgggccctggaacgc 1380 gcggtgggcatctga 1395 3-76 Sequences 3-76-1 SequenceNumber[ID] 76 3-76-2 MoleculeType AA 3-76-3 Length 464 source1...464 3-76-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Rhodococcusopacus NonEnglishQualifierValue 3-76-5 Residues MPLPMSPLDSMFLLGESREHPMHVGGVEIFQLPEGADTYDMRAMLDRALADGDGIVTPRL 60 AKRARRSFSSLGQWSWETVDDIDLGHHIRHDALPAPGGEAELMALCSRLHGSLLDRSRPL 120 WEMHLIEGLSDGRFAVYTKIHHAVADGVTAMKMLRNALSENSDDRDVPAPWQPRGPRPQR 180 TPSSKGFSLSGLAGSTLRTARETVGEVAGLVPALAGTVSRAFRDQGGPLALSAPKTPFNV 240 PITGARQFAAQSWPLERLRLVAKLSDSTINDVVLAMSSGALRSYLEDQNALPADPLIAMV 300 PVSLKSQREAATGNNIGVLMCNLGTHLREPADRLETIRTSMREGKEAYGSMTATQILAMS 360 ALGAAPIGASMLFGHNSRVRPPFNLIISNVPGPSSPLYWNGARLDAIYPLSVPVDGQGLN 420 ITCTSNDDIISFGVTGCRSAVPDLKSIPARLGHELRALERAVGI 464 3-77 Sequences 3-77-1 SequenceNumber[ID] 77 3-77-2 MoleculeType DNA 3-77-3 Length 1407 source1...1407 3-77-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Rhodococcusopacus NonEnglishQualifierValue 3-77-5 Residues atggccccgaccgactccctgttcctgctgggcgaatcccgcgagcacccgatgcacgtg 60 ggcggcctggcggtcttcaccccggcggagggcagctcggccgcggacgtccgcgccatg 120 ttcgacgccgcgctggtcggcgaccgggtggccgcgccgttccgcaagcgggcccgccgg 180 agcgtgacctcgctgggccagtggggctgggacaccctgcgcgacgacgaggtcgacctg 240 gaacaccatgtgcgccgggacgccctgccgcagccgggtggcatggcggaactgatgacc 300 ctggtctcccgcctgcatggcaccctgctggaccgcagccggccgctgtgggagatgcac 360 ctgatcgaaggcctggccgacggccggtacgcggtgtataccaagatccaccatgccctg 420 gcggacggcgccagcgcgatgcgcctgctgcgggactcgatgtccgaggacccgcatcgc 480 cggaacatgccgaccccctggcagccgcgcaaccccctgtcggccgtcccggacgccggc 540 gtcgcggtgacccccggccccggcagcgccctgcccgcgatggcctgggacgccgcgcgc 600 tccgccgcgggcgaagtcgccggcctgctgccggccgcgctgggcaccgtggaccgggcc 660 ctgcacggcaagggcggcgccctgtccctgaccgcgccgcataccctgttcaacgtcccc 720 atcagcggcgcccgccacgtggccgcgcggtcgttcccgatcgagcgcatccggctgctg 780 gccaagcatgccgacgcgaccatcaacgacatcgtgctgaccatgtgcgccggcaccctg 840 cgcgcgtacctgcacacccgcgacgccctgccggacaaccccctgatcgcgatggtcccg 900 gtgagcctgcgcgcccccgaaaccggcaccggcgaccgcgcccctggcggcaaccgggtc 960 ggcgtgctgatgtgcaacctggccacccacctgccggaccccgcgcatcgcctggagacc 1020 gtccggaactgcatgaacgaaggcaaggccgcgctgcaggccatgtcgccggcgcaggtc 1080 ctggccatgtccgcgctgggcgccgcgccgctgggcgtggagatgttcctgggccgccgg 1140 ggccccctgcgcccgcccttcaacgtcgtgatgtcgaacgtggcgggcccgcgcaccccc 1200 ctgtactggaacggcgcccggctggaatccctgtatccgctgagcatccccaccaccggc 1260 caggccctgaacatcacctgcacctccagcgacgaccagatcgtcttcggcctgaccggc 1320 tgccgccggaccgtgccggacctgcaccccatgctggaccagctggacgcggagctggac 1380 ctgctggaaaccgcggtcggcctgtga 1407 3-78 Sequences 3-78-1 SequenceNumber[ID] 78 3-78-2 MoleculeType AA 3-78-3 Length 468 source1...468 3-78-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Rhodococcusopacus NonEnglishQualifierValue 3-78-5 Residues MAPTDSLFLLGESREHPMHVGGLAVFTPAEGSSAADVRAMFDAALVGDRVAAPFRKRARR 60 SVTSLGQWGWDTLRDDEVDLEHHVRRDALPQPGGMAELMTLVSRLHGTLLDRSRPLWEMH 120 LIEGLADGRYAVYTKIHHALADGASAMRLLRDSMSEDPHRRNMPTPWQPRNPLSAVPDAG 180 VAVTPGPGSALPAMAWDAARSAAGEVAGLLPAALGTVDRALHGKGGALSLTAPHTLFNVP 240 ISGARHVAARSFPIERIRLLAKHADATINDIVLTMCAGTLRAYLHTRDALPDNPLIAMVP 300 VSLRAPETGTGDRAPGGNRVGVLMCNLATHLPDPAHRLETVRNCMNEGKAALQAMSPAQV 360 LAMSALGAAPLGVEMFLGRRGPLRPPFNVVISNVAGPRTPLYWNGARLESLYPLSIPTTG 420 QALNITCTSSDDQIVFGLTGCRRTVPDLHPMLDQLDAELDLLETAVGL 468 3-79 Sequences 3-79-1 SequenceNumber[ID] 79 3-79-2 MoleculeType DNA 3-79-3 Length 11299 misc_feature1...11299 3-79-4-1 FeaturesLocation/ note=pCM132 Qualifiers NonEnglishQualifierValue source1...11299 3-79-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-79-5 Residues gaccctttccgacgctcaccgggctggttgccctcgccgctgggctggcggccgtctatg 60 gccctgcaaacgcgccagaaacgccgtcgaagccgtgtgcgagacaccgcggccgccggc 120 gttgtggatacctcgcggaaaacttggccctcactgacagatgaggggcggacgttgaca 180 cttgaggggccgactcacccggcgcggcgttgacagatgaggggcaggctcgatttcggc 240 cggcgacgtggagctggccagcctcgcaaatcggcgaaaacgcctgattttacgcgagtt 300 tcccacagatgatgtggacaagcctggggataagtgccctgcggtattgacacttgaggg 360 gcgcgactactgacagatgaggggcgcgatccttgacacttgaggggcagagtgctgaca 420 gatgaggggcgcacctattgacatttgaggggctgtccacaggcagaaaatccagcattt 480 gcaagggtttccgcccgtttttcggccaccgctaacctgtcttttaacctgcttttaaac 540 caatatttataaaccttgtttttaaccagggctgcgccctgtgcgcgtgaccgcgcacgc 600 cgaaggggggtgcccccccttctcgaaccctcccggcccgctaacgcgggcctcccatcc 660 ccccaggggctgcgcccctcggccgcgaacggcctcaccccaaaaatggcagccaagctg 720 accacttctgcgctcggcccttccggctggctggtttattgctgataaatctggagccgg 780 tgagcgtgggtctcgcggtatcattgcagcactggggccagatggtaagccctcccgtat 840 cgtagttatctacacgacggggagtcaggcaactatggatgaacgaaatagacagatcgc 900 tgagataggtgcctcactgattaagcattggtaactgtcagaccaagtttactcatatat 960 actttagattgatttaaaacttcatttttaatttaaaaggatctaggtgaagatcctttt 1020 tgataatctcatgaccaaaatcccttaacgtgagttttcgttccactgagcgtcagaccc 1080 cgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgctt 1140 gcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaac 1200 tctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgtccttctagt 1260 gtagccgtagttaggccaccacttcaagaactctgtagcaccgcctacatacctcgctct 1320 gctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttgga 1380 ctcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcac 1440 acagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagctatg 1500 agaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggt 1560 cggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcc 1620 tgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcg 1680 gagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggcc 1740 ttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgc 1800 ctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgag 1860 cgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattca 1920 ttaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaat 1980 taatgtgagttagctcactcattaggcaccccaggctttacactttatgcttccggctcg 2040 tatgttgtgtggaattgtgagcggataacaatttcacacaggaaacagctatgaccatga 2100 ttacgccaagcttgcatgcctgcaggtcgactctagaggatcaattcggcttccaccgga 2160 attagcttgaaatagtacataatggatttccttacgcgaaatacgggcagacatggcctg 2220 cccggttattattatttttgacaccagaccaactggtaatggtagcgaccggcgctcagc 2280 tgtaattccgccgatactgacgggctccaggagtcgtcgccaccaatccccatatggaaa 2340 ccgtcgatattcagccatgtgccttcttccgcgtgcagcagatggcgatggctggtttcc 2400 atcagttgctgttgactgtagcggctgatgttgaactggaagtcgccgcgccactggtgt 2460 gggccataattcaattcgcgcgtcccgcagcgcagaccgttttcgctcgggaagacgtac 2520 ggggtatacatgtctgacaatggcagatcccagcggtcaaaacaggcggcagtaaggcgg 2580 tcgggatagttttcttgcggccctaatccgagccagtttacccgctctgctacctgcgcc 2640 agctggcagttcaggccaatccgcgccggatgcggtgtatcgctcgccacttcaacatca 2700 acggtaatcgccatttgaccactaccatcaatccggtaggttttccggctgataaataag 2760 gttttcccctgatgctgccacgcgtgagcggtcgtaatcagcaccgcatcagcaagtgta 2820 tctgccgtgcactgcaacaacgctgcttcggcctggtaatggcccgccgccttccagcgt 2880 tcgacccaggcgttagggtcaatgcgggtcgcttcacttacgccaatgtcgttatccagc 2940 ggtgcacgggtgaactgatcgcgcagcggcgtcagcagttgttttttatcgccaatccac 3000 atctgtgaaagaaagcctgactggcggttaaattgccaacgcttattacccagctcgatg 3060 caaaaatccatttcgctggtggtcagatgcgggatggcgtgggacgcggcggggagcgtc 3120 acactgaggttttccgccagacgccactgctgccaggcgctgatgtgcccggcttctgac 3180 catgcggtcgcgttcggttgcactacgcgtactgtgagccagagttgcccggcgctctcc 3240 ggctgcggtagttcaggcagttcaatcaactgtttaccttgtggagcgacatccagaggc 3300 acttcaccgcttgccagcggcttaccatccagcgccaccatccagtgcaggagctcgtta 3360 tcgctatgacggaacaggtattcgctggtcacttcgatggtttgcccggataaacggaac 3420 tggaaaaactgctgctggtgttttgcttccgtcagcgctggatgcggcgtgcggtcggca 3480 aagaccagaccgttcatacagaactggcgatcgttcggcgtatcgccaaaatcaccgccg 3540 taagccgaccacgggttgccgttttcatcatatttaatcagcgactgatccacccagtcc 3600 cagacgaagccgccctgtaaacggggatactgacgaaacgcctgccagtatttagcgaaa 3660 ccgccaagactgttacccatcgcgtgggcgtattcgcaaaggatcagcgggcgcgtctct 3720 ccaggtagcgaaagccattttttgatggaccatttcggcacagccgggaagggctggtct 3780 tcatccacgcgcgcgtacatcgggcaaataatatcggtggccgtggtgtcggctccgccg 3840 ccttcatactgcaccgggcgggaaggatcgacagatttgatccagcgatacagcgcgtcg 3900 tgattagcgccgtggcctgattcattccccagcgaccagatgatcacactcgggtgatta 3960 cgatcgcgctgcaccattcgcgttacgcgttcgctcatcgccggtagccagcgcggatca 4020 tcggtcagacgattcattggcaccatgccgtgggtttcaatattggcttcatccaccaca 4080 tacaggccgtagcggtcgcacagcgtgtaccacagcggatggttcggataatgcgaacag 4140 cgcacggcgttaaagttgttctgcttcatcagcaggatatcctgcaccatcgtctgctca 4200 tccatgacctgaccatgcagaggatgatgctcgtgacggttaacgcctcgaatcagcaac 4260 ggcttgccgttcagcagcagcagaccattttcaatccgcacctcgcggaaaccgacatcg 4320 caggcttctgcttcaatcagcgtgccgtcggcggtgtgcagttcaaccaccgcacgatag 4380 agattcgggatttcggcgctccacagtttcgggttttcgacgttcagacgtagtgtgacg 4440 cgatcggcataaccaccacgctcatcgataatttcaccgccgaaaggcgcggtgccgctg 4500 gcgacctgcgtttcaccctgccataaagaaactgttacccgtaggtagtcacgcaactcg 4560 ccgcacatctgaacttcagcctccagtacagcgcggctgaaatcatcattaaagcgagtg 4620 gcaacatggaaatcgctgatttgtgtagtcggtttatgcagcaacgagacgtcacggaaa 4680 atgccgctcatccgccacatatcctgatcttccagataactgccgtcactccaacgcagc 4740 accatcaccgcgaggcggttttctccggcgcgtaaaaatgcgctcaggtcaaattcagac 4800 ggcaaacgactgtcctggccgtaaccgacccagcgcccgttgcaccacagatgaaacgcc 4860 gagttaacgccatcaaaaataattcgcgtctggccttcctgtagccagctttcatcaaca 4920 ttaaatgtgagcgagtaacaacccgtcggattctccgtgggaacaaacggcggattgacc 4980 gtaatgggataggttacgttggtgtagatgggcgcatcgtaaccgtgcatctgccagttt 5040 gaggggacgacgacagtatcggcctcaggaagatcgcactccagccagctttccggcacc 5100 gcttctggtgccggaaaccaggcaaagcgccattcgccattcaggctgcgcaactgttgg 5160 gaagggcgatcggtgcgggcctcttcgctattacgccagctggcgaaagggggatgtgct 5220 gcaaggcgattaagttgggtaacgccagggttttcccggtcgacccgtaatcttacgtca 5280 gtaacttccacagtagttcaccaccttttccctatagatcttccgtgcagtttaagccga 5340 attgatccccgggtaccgagctcgaatctagaattccctgctttcctgatgcaaaaacga 5400 ggctagtttaccgtatctgtggggggatggcttgtagatatgacgacaggaagagtttgt 5460 agaaacgcaaaaaggccatccgtcaggatggccttctgcttaatttgatgcctggcagtt 5520 tatggcgggcgtcctgcccgccaccctccgggccgttgcttcgcaacgttcaaatccgct 5580 cccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaag 5640 gcccagtctttcgactgagcctttcgttttatttgatgcctggcagttccctactctcgc 5700 atggggagaccccacactaccatcggcgctacggcgtttcacttctgagttcggcatggg 5760 gtcaggtgggaccaccgcgctactgccgccaggcaaattctgttttatcagaccgcttct 5820 gcgttctgatttaatctgtatcaggctgaaaaattcactggccgtcgttttacaacgtcg 5880 tgactgggaaaaccctggcgttacccaacttaatcgccttgcagcacatccccctttcgc 5940 cagctggcgtaatagcgaagaggcccgcaccgatcgcccttcccaacagttgcgcagcct 6000 gaatggcgaatggcgcctgatgcggtattttctccttacgcatctgtgcggtatttcaca 6060 ccgcatatggtgcactctcagtacaatctgctctgatgccgcatagttaagccagccccg 6120 acacccgccaacacccgctgacgcgccctgacgggcttgtctgctcccggcatccgctta 6180 cagacaagctgtgaccgtctccgggagctgcatgtgtcagaggttttcaccgtcatcacc 6240 gaaacgcgcgagacgaaagggcctcgtgatacgcctatttttataggttaatgtcatgat 6300 aataatggtttcttagcaccctttctcggtccttcaacgttcctgacaacgagcctcctt 6360 ttcgccaatccatcgacaatcaccgcgagtccctgctcgaacgctgcgtccggaccggct 6420 tcgtcgaaggcgtctatcgcggcccgcaacagcggcgagagcggagcctgttcaacggtg 6480 ccgccgcgctcgccggcatcgctgtcgccggcctgctcctcaagcacggccccaacagtg 6540 aagtagctgattgtcatcagcgcattgacggcgtccccggccgaaaaacccgcctcgcag 6600 aggaagcgaagctgcgcgtcggccgtttccatctgcggtgcgcccggtcgcgtgccggca 6660 tggatgcgcgcgccatcgcggtaggcgagcagcgcctgcctgaagctgcgggcattcccg 6720 atcagaaatgagcgccagtcgtcgtcggctctcggcaccgaatgcgtatgattctccgcc 6780 agcatggcttcggccagtgcgtcgagcagcgcccgcttgttcctgaagtgccagtaaagc 6840 gccggctgctgaacccccaaccgttccgccagtttgcgtgtcgtcagaccgtctacgccg 6900 acctcgttcaacaggtccagggcggcacggatcactgtattcggctgcaactttgtcatg 6960 attgacactttatcactgataaacataatatgtccaccaacttatcagtgataaagaatc 7020 cgcgcgttcaatcggaccagcggaggctggtccggaggccagacgtgaaacccaacatac 7080 ccctgatcgtaattctgagcactgtcgcgctcgacgctgtcggcatcggcctgattatgc 7140 cggtgctgccgggcctcctgcgcgatctggttcactcgaacgacgtcaccgcccactatg 7200 gcattctgctggcgctgtatgcgttggtgcaatttgcctgcgcacctgtgctgggcgcgc 7260 tgtcggatcgtttcgggcggcggccaatcttgctcgtctcgctggccggcgccactgtcg 7320 actacgccatcatggcgacagcgcctttcctttgggttctctatatcgggcggatcgtgg 7380 ccggcatcaccggggcgactggggcggtagccggcgcttatattgccgatgacctgcagg 7440 ggggggggggcgctgaggtctgcctcgtgaagaaggtgttgctgactcataccaggcctg 7500 aatcgccccatcatccagccagaaagtgagggagccacggttgatgagagctttgttgta 7560 ggtggaccagttggtgattttgaacttttgctttgccacggaacggtctgcgttgtcggg 7620 aagatgcgtgatctgatccttcaactcagcaaaagttcgatttattcaacaaagccgccg 7680 tcccgtcaagtcagcgtaatgctctgccagtgttacaaccaattaaccaattctgattag 7740 aaaaactcatcgagcatcaaatgaaactgcaatttattcatatcaggattatcaatacca 7800 tatttttgaaaaagccgtttctgtaatgaaggagaaaactcaccgaggcagttccatagg 7860 atggcaagatcctggtatcggtctgcgattccgactcgtccaacatcaatacaacctatt 7920 aatttcccctcgtcaaaaataaggttatcaagtgagaaatcaccatgagtgacgactgaa 7980 tccggtgagaatggcaaaagcttatgcatttctttccagacttgttcaacaggccagcca 8040 ttacgctcgtcatcaaaatcactcgcatcaaccaaaccgttattcattcgtgattgcgcc 8100 tgagcgagacgaaatacgcgatcgctgttaaaaggacaattacaaacaggaatcgaatgc 8160 aaccggcgcaggaacactgccagcgcatcaacaatattttcacctgaatcaggatattct 8220 tctaatacctggaatgctgttttcccggggatcgcagtggtgagtaaccatgcatcatca 8280 ggagtacggataaaatgcttgatggtcggaagaggcataaattccgtcagccagtttagt 8340 ctgaccatctcatctgtaacatcattggcaacgctacctttgccatgtttcagaaacaac 8400 tctggcgcatcgggcttcccatacaatcgatagattgtcgcacctgattgcccgacatta 8460 tcgcgagcccatttatacccatataaatcagcatccatgttggaatttaatcgcggcctc 8520 gagcaagacgtttcccgttgaatatggctcataacaccccttgtattactgtttatgtaa 8580 gcagacagttttattgttcatgatgatatatttttatcttgtgcaatgtaacatcagaga 8640 ttttgagacacaacgtggctttcccccccccccctgcaggtccgacacggggatggatgg 8700 cgttcccgatcatggtcctgcttgcttcgggtggcatcggaatgccggcgctgcaagcaa 8760 tgttgtccaggcaggtggatgaggaacgtcaggggcagctgcaaggctcactggcggcgc 8820 tcaccagcctgacctcgatcgtcggacccctcctcttcacggcgatctatgcggcttcta 8880 taacaacgtggaacgggtgggcatggattgcaggcgctgccctctacttgctctgcctgc 8940 cggcgctgcgtcgcgggctttggagcggcgcagggcaacgagccgatcgctgatcgtgga 9000 aacgataggcctatgccatgcgggtcaaggcgacttccggcaagctatacgcgccctaga 9060 attgtcaattttaatcctctgtttatcggcagttcgtagagcgcgccgtgcgtcccgagc 9120 gatactgagcgaagcaagtgcgtcgagcagtgcccgcttgttcctgaaatgccagtaaag 9180 cgctggctgctgaacccccagccggaactgaccccacaaggccctagcgtttgcaatgca 9240 ccaggtcatcattgacccaggcgtgttccaccaggccgctgcctcgcaactcttcgcagg 9300 cttcgccgacctgctcgcgccacttcttcacgcgggtggaatccgatccgcacatgaggc 9360 ggaaggtttccagcttgagcgggtacggctcccggtgcgagctgaaatagtcgaacatcc 9420 gtcgggccgtcggcgacagcttgcggtacttctcccatatgaatttcgtgtagtggtcgc 9480 cagcaaacagcacgacgatttcctcgtcgatcaggacctggcaacgggacgttttcttgc 9540 cacggtccaggacgcggaagcggtgcagcagcgacaccgattccaggtgcccaacgcggt 9600 cggacgtgaagcccatcgccgtcgcctgtaggcgcgacaggcattcctcggccttcgtgt 9660 aataccggccattgatcgaccagcccaggtcctggcaaagctcgtagaacgtgaaggtga 9720 tcggctcgccgataggggtgcgcttcgcgtactccaacacctgctgccacaccagttcgt 9780 catcgtcggcccgcagctcgacgccggtgtaggtgatcttcacgtccttgttgacgtgga 9840 aaatgaccttgttttgcagcgcctcgcgcgggattttcttgttgcgcgtggtgaacaggg 9900 cagagcgggccgtgtcgtttggcatcgctcgcatcgtgtccggccacggcgcaatatcga 9960 acaaggaaagctgcatttccttgatctgctgcttcgtgtgtttcagcaacgcggcctgct 10020 tggcctcgctgacctgttttgccaggtcctcgccggcggtttttcgcttcttggtcgtca 10080 tagttcctcgcgtgtcgatggtcatcgacttcgccaaacctgccgcctcctgttcgagac 10140 gacgcgaacgctccacggcggccgatggcgcgggcagggcagggggagccagttgcacgc 10200 tgtcgcgctcgatcttggccgtagcttgctggaccatcgagccgacggactggaaggttt 10260 cgcggggcgcacgcatgacggtgcggcttgcgatggtttcggcatcctcggcggaaaacc 10320 ccgcgtcgatcagttcttgcctgtatgccttccggtcaaacgtccgattcattcaccctc 10380 cttgcgggattgccccgactcacgccggggcaatgtgcccttattcctgatttgacccgc 10440 ctggtgccttggtgtccagataatccaccttatcggcaatgaagtcggtcccgtagaccg 10500 tctggccgtccttctcgtacttggtattccgaatcttgccctgcacgattaccagctccg 10560 cgaagtcgctcttcttgatggagcgcatggggacgtgcttggcaatcacgcgcacccccc 10620 ggccgttttagcggctaaaaaagtcatggctctgccctcgggcggaccacgcccatcatg 10680 accttgccaagctcgtcctgcttctcttcgatcttcgccagcagggcgaggatcgtggca 10740 tcaccgaaccgcgccgtgcgcgggtcgtcggtgagccagagtttcagcaggccgcccagg 10800 cggcccaggtcgccattgatgcgggccagctcgcggacgtgctcatagtccacgacgccc 10860 gtgattttgtagccctggccgacggccagcaggtaggcctacaggctcatgccggccgcc 10920 gccgccttttcctcaatcgctcttcgttcgtctggaaggcagtacaccttgataggtggg 10980 ctgcccttcctggttggcttggtttcatcagccatccgcttgccctcatctgttacgccg 11040 gcggtagccggccagcctcgcagagcaggattcccgttgagcaccgccaggtgcgaataa 11100 gggacagtgaagaaggaacacccgctcgcgggtgggcctacttcacctatcctgcccggc 11160 tgacgccgttggatacaccaaggaaagtctacacgaaccctttggcaaaatcctgtatat 11220 cgtgcgaaaaaggatggatataccgaaaaaatcgctataatgaccccgaagcagggttat 11280 gcagcggaaaagatccgtc 11299 3-80 Sequences 3-80-1 SequenceNumber[ID] 80 3-80-2 MoleculeType DNA 3-80-3 Length 7341 3-80-4-1 FeaturesLocation/ misc_feature1...7341 Qualifiers note=pJSvec NonEnglishQualifierValue source1...7341 3-80-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-80-5 Residues gaccctttccgacgctcaccgggctggttgccctcgccgctgggctggcggccgtctatg 60 gccctgcaaacgcgccagaaacgccgtcgaagccgtgtgcgagacaccgcggccgccggc 120 gttgtggatacctcgcggaaaacttggccctcactgacagatgaggggcggacgttgaca 180 cttgaggggccgactcacccggcgcggcgttgacagatgaggggcaggctcgatttcggc 240 cggcgacgtggagctggccagcctcgcaaatcggcgaaaacgcctgattttacgcgagtt 300 tcccacagatgatgtggacaagcctggggataagtgccctgcggtattgacacttgaggg 360 gcgcgactactgacagatgaggggcgcgatccttgacacttgaggggcagagtgctgaca 420 gatgaggggcgcacctattgacatttgaggggctgtccacaggcagaaaatccagcattt 480 gcaagggtttccgcccgtttttcggccaccgctaacctgtcttttaacctgcttttaaac 540 caatatttataaaccttgtttttaaccagggctgcgccctgtgcgcgtgaccgcgcacgc 600 cgaaggggggtgcccccccttctcgaaccctcccggcccgctaacgcgggcctcccatcc 660 ccccaggggctgcgcccctcggccgcgaacggcctcaccccaaaaatggcagccaagctg 720 accacttctgcgctcggcccttccggctggctggtttattgctgataaatctggagccgg 780 tgagcgtgggtctcgcggtatcattgcagcactggggccagatggtaagccctcccgtat 840 cgtagttatctacacgacggggagtcaggcaactatggatgaacgaaatagacagatcgc 900 tgagataggtgcctcactgattaagcattggtaactgtcagaccaagtttactcatatat 960 actttagattgatttaaaacttcatttttaatttaaaaggatctaggtgaagatcctttt 1020 tgataatctcatgaccaaaatcccttaacgtgagttttcgttccactgagctgcagaccc 1080 cgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgctt 1140 gcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaac 1200 tctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgtccttctagt 1260 gtagccgtagttaggccaccacttcaagaactctgtagcaccgcctacatacctcgctct 1320 gctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttgga 1380 ctcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcac 1440 acagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagctatg 1500 agaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggt 1560 cggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcc 1620 tgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcg 1680 gagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggcc 1740 ttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgc 1800 ctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgag 1860 cgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattca 1920 ttaatgcagctggcaggaagcggcgatggcggagctgaattacattcccaaccgcgtggc 1980 acaacaactggcgggcaaacagtcgttgctgattggcgttgccacctccagtctggccct 2040 gcacgcgccgtcgcaaattgtcgcggcgattaaatctcgcgccgatcaactgggtgccag 2100 cgtggtggtgtcgatggtagaacgaagcggcgtcgaagcctgtaaagcggcggtgcacaa 2160 tcttctcgcgcaacgcgtcagtgggctgatcattaactatccgctggatgaccaggatgc 2220 cattgctgtggaagctgcctgcactaatgttccggcgttatttcttgatgtctctgacca 2280 gacacccatcaacagtattattttctcccatgaagacggtacgcgactgggcgtggagca 2340 tctggtcgcattgggtcaccagcaaatcgcgctgttagcgggcccattaagttctgtctc 2400 ggcgcgtctgcgtctggctggctggcataaatatctcactcgcaatcaaattcagccgat 2460 agcggaacgggaaggcgactggagtgccatgtccggttttcaacaaaccatgcaaatgct 2520 gaatgagggcatcgttcccactgcgatgctggttgccaacgatcagatggcgctgggcgc 2580 aatgcgcgccattaccgagtccgggctgcgcgttggtgcggatatctcggtagtgggata 2640 cgacgataccgaagacagctcatgttatatcccgccgtcaaccaccatcaaacaggattt 2700 tcgcctgctggggcaaaccagcgtggaccgcttgctgcaactctctcagggccaggcggt 2760 gaagggcaatcagctgttgcccgtctcactggtgaaaagaaaaaccaccctggcgcccaa 2820 tacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggt 2880 ttcccgactggaaagcgggcagtgagcgcaacgcaattaatgtgagttagcgcgaattga 2940 tctggtttgacagcttatcatcgactgcacggtgcaccaatgcttctggcgtcaggcagc 3000 catcggaagctgtggtatggctgtgcaggtctgaaatcactgcataattcgtgtcgctca 3060 aggcgcactcccgttctggataatgttttttgcgccgacatcataacggttctggcaaat 3120 attctgaaatgagctgttgacaattaatcatccggctcgtataatgtgtggaattgtgag 3180 cggataacaatttcacacaggaaacagcgccgctgagaaaaagcgaagcggcactgctct 3240 ttaacaatttatcagacaatctgtgtgggcactcgaccggaattatcgattaactttatt 3300 attaaaaattaaagaggtatatattaatgtatcgattaaataaggaggaataaacccaga 3360 acgcagaagcggtctgataaaacagaatttgcctggcggcagtagcgcggtggtcccacc 3420 tgaccccatgccgaactcagaagtgaaacgccgtagcgccgatggtagtgtggggtctcc 3480 ccatgcgagagtagggaactgccaggcatcaaataaaacgaaaggctcagtcgaaagact 3540 gggcctttcgttttatctgttgtttgtcggtgaacgctctcctgagtaggacaaatcttg 3600 taggtggaccagttggtgattttgaacttttgctttgccacggaacggtctgcgttgtcg 3660 ggaagatgcgtgatctgatccttcaactcagcaaaagttcgatttattcaacaaagccgc 3720 cgtcccgtcaagtcagcgtaatgctctgccagtgttacaaccaattaaccaattctgatt 3780 agaaaaactcatcgagcatcaaatgaaactgcaatttattcatatcaggattatcaatac 3840 catatttttgaaaaagccgtttctgtaatgaaggagaaaactcaccgaggcagttccata 3900 ggatggcaagatcctggtatcggtctgcgattccgactcgtccaacatcaatacaaccta 3960 ttaatttcccctcgtcaaaaataaggttatcaagtgagaaatcaccatgagtgacgactg 4020 aatccggtgagaatggcaaaagcttatgcatttctttccagacttgttcaacaggccagc 4080 cattacgctcgtcatcaaaatcactcgcatcaaccaaaccgttattcattcgtgattgcg 4140 cctgagcgagacgaaatacgcgatcgctgttaaaaggacaattacaaacaggaatcgaat 4200 gcaaccggcgcaggaacactgccagcgcatcaacaatattttcacctgaatcaggatatt 4260 cttctaatacctggaatgctgttttcccggggatcgcagtggtgagtaaccatgcatcat 4320 caggagtacggataaaatgcttgatggtcggaagaggcataaattccgtcagccagttta 4380 gtctgaccatctcatctgtaacatcattggcaacgctacctttgccatgtttcagaaaca 4440 actctggcgcatcgggcttcccatacaatcgatagattgtcgcacctgattgcccgacat 4500 tatcgcgagcccatttatacccatataaatcagcatccatgttggaatttaatcgcggcc 4560 tcgagcaagacgtttcccgttgaatatggctcataacaccccttgtattactgtttatgt 4620 aagcagacagttttattgttcatgatgatatatttttatcttgtgcaatgtaacatcaga 4680 gattttgagacacaacgtggctttcccccccccccctgcaggtccgacacggggatggat 4740 ggcgttcccgatcatggtcctgcttgcttcgggtggcatcggaatgccggcgctgcaagc 4800 aatgttgtccaggcaggtggatgaggaacgtcaggggcagctgcaaggctcactggcggc 4860 gctcaccagcctgacctcgatcgtcggacccctcctcttcacggcgatctatgcggcttc 4920 tataacaacgtggaacgggtgggcatggattgcaggcgctgccctctacttgctctgcct 4980 gccggcgctgcgtcgcgggctttggagcggcgcagggcaacgagccgatcgctgatcgtg 5040 gaaacgataggcctatgccatgcgggtcaaggcgacttccggcaagctatacgcgcccta 5100 gaattgtcaattttaatcctctgtttatcggcagttcgtagagcgcgccgtgcgtcccga 5160 gcgatactgagcgaagcaagtgcgtcgagcagtgcccgcttgttcctgaaatgccagtaa 5220 agcgctggctgctgaacccccagccggaactgaccccacaaggccctagcgtttgcaatg 5280 caccaggtcatcattgacccaggcgtgttccaccaggccgctgcctcgcaactcttcgca 5340 ggcttcgccgacctgctcgcgccacttcttcacgcgggtggaatccgatccgcacatgag 5400 gcggaaggtttccagcttgagcgggtacggctcccggtgcgagctgaaatagtcgaacat 5460 ccgtcgggccgtcggcgacagcttgcggtacttctcccatatgaatttcgtgtagtggtc 5520 gccagcaaacagcacgacgatttcctcgtcgatcaggacctggcaacgggacgttttctt 5580 gccacggtccaggacgcggaagcggtgcagcagcgacaccgattccaggtgcccaacgcg 5640 gtcggacgtgaagcccatcgccgtcgcctgtaggcgcgacagccattcctcggccttcgt 5700 gtaataccggccattgatcgaccagcccaggtcctggcaaagctcgtagaacgtgaaggt 5760 gatcggctcgccgataggggtgcgcttcgcgtactccaacacctgctgccacaccagttc 5820 gtcatcgtcggcccgcagctcgacgccggtgtaggtgatcttcacgtccttgttgacgtg 5880 gaaaatgaccttgttttgcagcgcctcgcgcgggattttcttgttgcgcgtggtgaacag 5940 ggcagagcgggccgtgtcgtttggcatcgctcgcatcgtgtccggccacggcgcaatatc 6000 gaacaaggaaagctgcatttccttgatctgctgcttcgtgtgtttcagcaacgcggcctg 6060 cttggcctcgctgacctgttttgccaggtcctcgccggcggtttttcgcttcttggtcgt 6120 catagttcctcgcgtgtcgatggtcatcgacttcgccaaacctgccgcctcctgttcgag 6180 acgacgcgaacgctccacggcggccgatggcgcgggcagggcagggggagccagttgcac 6240 gctgtcgcgctcgatcttggccgtagcttgctggaccatctcggcatcctacgtgaaggt 6300 ttcgcggggcgcacgcatgacggtgcggcttgcgatggttaacgtccgatcggcggaaaa 6360 ccccgcgtcgatcagttcttgcctgtatgccttccggtcaccttattccttcattcaccc 6420 tccttgcgggattgccccgactcacgccggggcaatgtgcatgaagtcgggatttgaccc 6480 gcctggtgccttggtgtccagataatccaccttatcggcaccctgcacgatcccgtagac 6540 cgtctggccgtccttctcgtacttggtattccgaatcttgttggcaatcaataccagctc 6600 cgcgaagtcgctcttcttgatggagcgcatggggacgtgccgggcggacccgcgcacccc 6660 ccggccgttttagcggctaaaaaagtcatggctctgccctcagcagggcgacgcccatca 6720 tgaccttgccaagctcgtcctgcttctcttcgatcttcgcgagtttcagcaggatcgtgg 6780 catcaccgaaccgcgccgtgcgcgggtcgtcggtgagccagtgctcatgcaggccgccca 6840 ggcggcccaggtcgccattgatgcgggccagctcgcggacgtgctcatagtccacgacgc 6900 ccgtgattttgtagccctggccgacggccagcaggtaggcctacaggctcatgccggccg 6960 ccgccgccttttcctcaatcgctcttcgttcgtctggaaggcagtacaccttgataggtg 7020 ggctgcccttcctggttggcttggtttcatcagccatccgcttgccctcatctgttacgc 7080 cggcggtagccggccagcctcgcagagcaggattcccgttgagcaccgccaggtgcgaat 7140 aagggacagtgaagaaggaacacccgctcgcgggtgggcctacttcacctatcctgcccg 7200 gctgacgccgttggatacaccaaggaaagtctacacgaaccctttggcaaaatcctgtat 7260 atcgtgcgaaaaaggatggatataccgaaaaaatcgctataatgaccccgaagcagggtt 7320 atgcagcggaaaagatccgtc 7341 3-81 Sequences 3-81-1 SequenceNumber[ID] 81 3-81-2 MoleculeType DNA 3-81-3 Length 10705 3-81-4-1 FeatureLocation/ misc_feature1...10705 Qualifiers note=pMZT3 NonEnglishQualifierValue source1...10705 3-81-4-2 FeatureLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-81-5 Residues tcaccctgtcgggcaatgccgaggcattctggcagcagcgccccctggcctgtagtggat 60 tacgtgccggtctgttccatcctaccaccggctattcactgccgctggcggttgccgtgg 120 ccgaccgcctgagcgcacttgatgtctttacgtcggcctcaattcaccaggctattaggc 180 attttgcccgcgagcgctggcagcagcagcgctttttccgcatgctgaatcgcatgctgt 240 ttttagccggacccgccgattcacgctggcgggttatgcagcgtttttatggtttacctg 300 aagatttaattgcccgtttttatgcgggaaaactcacgctgaccgatcggctacgtattc 360 tgagcggcaagccgcctgttccggtattagcagcattgcaagccattatgacgactcatc 420 gttaagagacagaacgaagtgtgaccagaacgcagaagcggtctgataaaacagaatttg 480 cctggcggcagtagcgcggtggtcccacctgaccccatgccgaactcagaagtgaaacgc 540 cgtagcgccgatggtagtgtggggtctccccatgcgagagtagggaactgccaggcatca 600 aataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggt 660 gaacgctctcctgagtaggacaaatcttgtaggtggaccagttggtgattttgaactttt 720 gctttgccacggaacggtctgcgttgtcgggaagatgcgtgatctgatccttcaactcag 780 caaaagttcgatttattcaacaaagccgccgtcccgtcaagtcagcgtaatgctctgcca 840 gtgttacaaccaattaaccaattctgattagaaaaactcatcgagcatcaaatgaaactg 900 caatttattcatatcaggattatcaataccatatttttgaaaaagccgtttctgtaatga 960 aggagaaaactcaccgaggcagttccataggatggcaagatcctggtatcggtctgcgat 1020 tccgactcgtccaacatcaatacaacctattaatttcccctcgtcaaaaataaggttatc 1080 aagtgagaaatcaccatgagtgacgactgaatccggtgagaatggcaaaagcttatgcat 1140 ttctttccagacttgttcaacaggccagccattacgctcgtcatcaaaatcactcgcatc 1200 aaccaaaccgttattcattcgtgattgcgcctgagcgagacgaaatacgcgatcgctgtt 1260 aaaaggacaattacaaacaggaatcgaatgcaaccggcgcaggaacactgccagcgcatc 1320 aacaatattttcacctgaatcaggatattcttctaatacctggaatgctgttttcccggg 1380 gatcgcagtggtgagtaaccatgcatcatcaggagtacggataaaatgcttgatggtcgg 1440 aagaggcataaattccgtcagccagtttagtctgaccatctcatctgtaacatcattggc 1500 aacgctacctttgccatgtttcagaaacaactctggcgcatcgggcttcccatacaatcg 1560 atagattgtcgcacctgattgcccgacattatcgcgagcccatttatacccatataaatc 1620 agcatccatgttggaatttaatcgcggcctcgagcaagacgtttcccgttgaatatggct 1680 cataacaccccttgtattactgtttatgtaagcagacagttttattgttcatgatgatat 1740 atttttatcttgtgcaatgtaacatcagagattttgagacacaacgtggctttccccccc 1800 ccccctgcaggtccgacacggggatggatggcgttcccgatcatggtcctgcttgcttcg 1860 ggtggcatcggaatgccggcgctgcaagcaatgttgtccaggcaggtggatgaggaacgt 1920 caggggcagctgcaaggctcactggcggcgctcaccagcctgacctcgatcgtcggaccc 1980 ctcctcttcacggcgatctatgcggcttctataacaacgtggaacgggtgggcatggatt 2040 gcaggcgctgccctctacttgctctgcctgccggcgctgcgtcgcgggctttggagcggc 2100 gcagggcaacgagccgatcgctgatcgtggaaacgataggcctatgccatgcgggtcaag 2160 gcgacttccggcaagctatacgcgccctagaattgtcaattttaatcctctgtttatcgg 2220 cagttcgtagagcgcgccgtgcgtcccgagcgatactgagcgaagcaagtgcgtcgagca 2280 gtgcccgcttgttcctgaaatgccagtaaagcgctggctgctgaacccccagccggaact 2340 gaccccacaaggccctagcgtttgcaatgcaccaggtcatcattgacccaggcgtgttcc 2400 accaggccgctgcctcgcaactcttcgcaggcttcgccgacctgctcgcgccacttcttc 2460 acgcgggtggaatccgatccgcacatgaggcggaaggtttccagcttgagcgggtacggc 2520 tcccggtgcgagctgaaatagtcgaacatccgtcgggccgtcggcgacagcttgcggtac 2580 ttctcccatatgaatttcgtgtagtggtcgccagcaaacagcacgacgatttcctcgtcg 2640 atcaggacctggcaacgggacgttttcttgccacggtccaggacgcggaagcggtgcagc 2700 agcgacaccgattccaggtgcccaacgcggtcggacgtgaagcccatcgccgtcgcctgt 2760 aggcgcgacaggcattcctcggccttcgtgtaataccggccattgatcgaccagcccagg 2820 tcctggcaaagctcgtagaacgtgaaggtgatcggctcgccgataggggtgcgcttcgcg 2880 tactccaacacctgctgccacaccagttcgtcatcgtcggcccgcagctcgacgccggtg 2940 taggtgatcttcacgtccttgttgacgtggaaaatgaccttgttttgcagcgcctcgcgc 3000 gggattttcttgttgcgcgtggtgaacagggcagagcgggccgtgtcgtttggcatcgct 3060 cgcatcgtgtccggccacggcgcaatatcgaacaaggaaagctgcatttccttgatctgc 3120 tgcttcgtgtgtttcagcaacgcggcctgcttggcctcgctgacctgttttgccaggtcc 3180 tcgccggcggtttttcgcttcttggtcgtcatagttcctcgcgtgtcgatggtcatcgac 3240 ttcgccaaacctgccgcctcctgttcgagacgacgcgaacgctccacggcggccgatggc 3300 gcgggcagggcagggggagccagttgcacgctgtcgcgctcgatcttggccgtagcttgc 3360 tggaccatcgagccgacggactggaaggtttcgcggggcgcacgcatgacggtgcggctt 3420 gcgatggtttcggcatcctcggcggaaaaccccgcgtcgatcagttcttgcctgtatgcc 3480 ttccggtcaaacgtccgattcattcaccctccttgcgggattgccccgactcacgccggg 3540 gcaatgtgcccttattcctgatttgacccgcctggtgccttggtgtccagataatccacc 3600 ttatcggcaatgaagtcggtcccgtagaccgtctggccgtccttctcgtacttggtattc 3660 cgaatcttgccctgcacgaataccagctccgcgaagtcgctcttcttgatggagcgcatg 3720 gggacgtgcttggcaatcacgcgcaccccccggccgttttagcggctaaaaaagtcatgg 3780 ctctgccctcgggcggaccacgcccatcatgaccttgccaagctcgtcctgcttctcttc 3840 gatcttcgccagcagggcgaggatcgtggcatcaccgaaccgcgccgtgcgcgggtcgtc 3900 ggtgagccagagtttcagcaggccgcccaggcggcccaggtcgccattgatgcgggccag 3960 ctcgcggacgtgctcatagtccacgacgcccgtgattttgtagccctggccgacggccag 4020 caggtaggcctacaggctcatgccggccgccgccgccttttcctcaatcgctcttcgttc 4080 gtctggaaggcagtacaccttgataggtgggctgcccttcctggttggcttggtttcatc 4140 agccatccgcttgccctcatctgttacgccggcggtagccggccagcctcgcagagcagg 4200 attcccgttgagcaccgccaggtgcgaataagggacagtgaagaaggaacacccgctcgc 4260 gggtgggcctacttcacctatcctgcccggctgacgccgttggatacaccaaggaaagtc 4320 tacacgaaccctttggcaaaatcctgtatatcgtgcgaaaaaggatggatataccgaaaa 4380 aatcgctataatgaccccgaagcagggttatgcagcggaaaagatccgtcgaccctttcc 4440 gacgctcaccgggctggttgccctcgccgctgggctggcggccgtctatggccctgcaaa 4500 cgcgccagaaacgccgtcgaagccgtgtgcgagacaccgcggccgccggcgttgtggata 4560 cctcgcggaaaacttggccctcactgacagatgaggggcggacgttgacacttgaggggc 4620 cgactcacccggcgcggcgttgacagatgaggggcaggctcgatttcggccggcgacgtg 4680 gagctggccagcctcgcaaatcggcgaaaacgcctgattttacgcgagtttcccacagat 4740 gatgtggacaagcctggggataagtgccctgcggtattgacacttgaggggcgcgactac 4800 tgacagatgaggggcgcgatccttgacacttgaggggcagagtgctgacagatgaggggc 4860 gcacctattgacatttgaggggctgtccacaggcagaaaatccagcatttgcaagggttt 4920 ccgcccgtttttcggccaccgctaacctgtcttttaacctgcttttaaaccaatatttat 4980 aaaccttgtttttaaccagggctgcgccctgtgcgcgtgaccgcgcacgccgaagggggg 5040 tgcccccccttctcgaaccctcccggcccgctaacgcgggcctcccatccccccaggggc 5100 tgcgcccctcggccgcgaacggcctcaccccaaaaatggcagccaagctgaccacttctg 5160 cgctcggcccttccggctggctggtttattgctgataaatctggagccggtgagcgtggg 5220 tctcgcggtatcattgcagcactggggccagatggtaagccctcccgtatcgtagttatc 5280 tacacgacggggagtcaggcaactatggatgaacgaaatagacagatcgctgagataggt 5340 gcctcactgattaagcattggtaactgtcagaccaagtttactcatatatactttagatt 5400 gatttaaaacttcatttttaatttaaaaggatctaggtgaagatcctttttgataatctc 5460 atgaccaaaatcccttaacgtgagttttcgttccactgagcgtcagaccccgtagaaaag 5520 atcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaa 5580 aaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccg 5640 aaggtaactggcttcagcagagcgcagataccaaatactgtccttctagtgtagccgtag 5700 ttaggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctg 5760 ttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacga 5820 tagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagc 5880 ttggagcgaacgacctacaccgaactgagatacctacagcgtgagctatgagaaagcgcc 5940 acgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacagga 6000 gagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggttt 6060 cgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatgg 6120 aaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcac 6180 atgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtga 6240 gctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcg 6300 gaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagc 6360 tggcagccagcgcttcgttaatacagatgtaggtgttccacagggtagccagcagcatcc 6420 tgcgatgcagatccggaacataatggtgcagggcgctgactttatagctagctcagccct 6480 tggtacaatgctagcgagcaaccaacacttaaagaggagaaaatgtatccgtttataagg 6540 acagcccgaatgacggtctgcgcaaaaaaacacgttcatctcactcgcgatgctgcggag 6600 cagttactggctgatattgatcgacgccttgatcagttattgcccgtggagggagaacgg 6660 gatgttgtgggtgccgcgatgcgtgaaggtgcgctggcaccgggaaaacgtattcgcccc 6720 atgttgctgttgctgaccgcccgcgatctgggttgcgctgtcagccatgacggattactg 6780 gatttggcctgtgcggtggaaatggtccacgcggcttcgctgatccttgacgatatgccc 6840 tgcatggacgatgcgaagctgcggcgcggacgccctaccattcattctcattacggagag 6900 catgtggcaatactggcggcggttgccttgctgagtaaagcctttggcgtaattgccgat 6960 gcagatggcctcacgccgctggcaaaaaatcgggcggtttctgaactgtcaaacgccatc 7020 ggcatgcaaggattggttcagggtcagttcaaggatctgtctgaaggggataagccgcgc 7080 agcgctgaagctattttgatgacgaatcactttaaaaccagcagcgtgttttgtgcctcc 7140 atgcagatggcctcgattgttgcgaatgcctccagcgaagcgcgtgattgcctgcatcgt 7200 ttttcacttgatcttggtcaggcatttcaactgctggacgatttgaccgatggcatgacc 7260 gacaccggtaaggatagcaatcaggacgccggtaaatcgacgctggtcaatctgttaggc 7320 cctagggcggttgaagaacgtctgagacaacatcttcatcttgccagtgagcatctctct 7380 gcggcctgccaacacgggcacgccactcaacattttattcaggcctggtttgacaaaaaa 7440 ctcgctgccgtcagttaaaggtctctagacaggatgtgtcacacaggaaaccatgaaacc 7500 aactacggtaattggtgcaggcttcggtggcctggcactggcaattcgtctacaggctgc 7560 ggggatccccgtcttactgcttgaacaacgtgataaacccggcggtcgggcttatgtcta 7620 cgaggatcaggggtttacctttgatgcaggcccgacggttatcaccgatcccagtgccat 7680 tgaagaactgtttgcactggcaggaaaacagttaaaagagtatgtcgaactgctgccggt 7740 tacgccgttttaccgcctgtgttgggagtcagggaaggtctttaattacgataacgatca 7800 aacccggctcgaagcgcagattcagcagtttaatccccgcgatgtcgaaggttatcgtca 7860 gtttctggactattcacgcgcggtgtttaaagaaggctatctgaagctcggtactgtccc 7920 ttttttatcgttcagagacatgcttcgcgccgcacctcaactggcgaaactgcaggcatg 7980 gagaagcgtttacagtaaggttgccagttacatcgaagatgaacatctgcgccaggcgtt 8040 ttctttccactcgctgttggtgggcggcaatcccttcgccacctcatccatttatacgtt 8100 gatacacgcgctggagcgtgagtggggcgtctggtttccgcgtggcggcaccggcgcatt 8160 agttcaggggatgataaagctgtttcaggatctgggtggtgaagtcgtgttaaacgccag 8220 agtcagccatatggaaacgacaggaaacaagattgaagccgtgcatttagaggacggtcg 8280 caggttcctgacgcaagccgtcgcgtcaaatgcagatgtggttcatacctatcgcgacct 8340 gttaagccagcaccctgccgcggttaagcagtccaacaaactgcagactaagcgtatgag 8400 taactctctgtttgtgctctattttggtttgaatcaccatcatgatcagctcgcgcatca 8460 cacggtttgtttcggcccgcgttaccgcgaactgattgacgagatttttaatcatgatgg 8520 cctcgcagaagacttctcactttatctgcacgcgccctgtgtcacggattcgtcactggc 8580 gcctgaaggttgcggcagttactatgtgttggcgccggtgccgcatttaggcaccgcgaa 8640 cctcgactggacggttgaggggccaaaactacgcgaccgtatttttgagtaccttgagca 8700 gcattacatgcctggcttacggagtcagctggtcacgcaccagatgtttacgccgtttga 8760 ttttcgcgaccagcttaatgcctatcagggctcagccttttctgtggagcccgttcttac 8820 ccagagcgcctggtttcggccgcataaccgcgataaaaccattactaatctctacctggt 8880 cggcgcaggcacgcatcccggcgcaggcattcctggcgtcatcggctcggcaaaagcgac 8940 agcaggtttgatgctggaggatctgatttaagtgatcgttgagtggtgaacttaaagagg 9000 agaaaatgaataatccgtcgttactcaatcatgcggtcgaaacgatggcagttggctcga 9060 aaagttttgcgacagcctcaaagttatttgatgcaaaaacccggcgcagcgtactgatgc 9120 tctacgcctggtgccgccattgtgacgatgttattgacgaccagacgctgggcttccagg 9180 cccggcagcctgccttacaaacgcccgaacaacgtctgatgcaacttgagatgaaaacgc 9240 gccaggcctatgcaggatcgcagatgcacgaaccggcgtttgcggcttttcaggaagtgg 9300 ctatggctcatgatatcgccccggcttacgcgtttgatcatctggaaggcttcgccatgg 9360 atgtacgcgaagcgcaatacagccaactggacgatacgctgcgctattgctatcacgttg 9420 caggcgttgtcggcttgatgatggcgcaaatcaggggcgtacgggataacgccacgctgg 9480 accgcgcctgtgaccttgggctggcatttcagttgaccaatattgctcgcgataggtggg 9540 acgatgcgcatgcgggccgctgttatctgccggcaagctggctggagcatgaaggtctga 9600 acaaagagaattatgcggcacctgaaaaccgtcaggcgctgagccgtatcgcccgtcgtt 9660 tggtgcaggaagcagaaccttactatttgtctgccacagcgggcctggctgggttgcccc 9720 tgcgttcggcctgggcaatcgctacggcgaagcaggtttaccggaaaataggtgtcaaag 9780 ttgaacaggccggtcagcaagcctgggatcagcggcagtcaacgaccacgcccgaaaaat 9840 taacgctgctgctggccgcctctggtcaggcccttacttcccggatgcgggctcatcctc 9900 cccgccctgcgcatctctggcagcgcccgctctaatcacgtagcaagctgacagtttaaa 9960 gaggagaaaatgggagcggctatgcaaccgcattatgatctgattctcgtgggggctgga 10020 ctcgcgaatggccttatcgccctgcgtcttcagcagcagcaacctgatatgcgtattttg 10080 cttatcgacgccgcaccccaggcgggcgggaatcatacgtggtcatttcaccacgatgat 10140 ttgactgagagccaacatcgttggatagcttcgctggtggttcatcactggcccgactat 10200 caggtacgctttcccacacgccgtcgtaagctgaacagcggctacttctgtattacttct 10260 cagcgtttcgctgaggttttacagcgacagtttggcccgcacttgtggatggataccgcg 10320 gtcgcagaggttaatgcggaatctgttcggttgaaaaagggtcaggttatcggtgcccgc 10380 gcggtgattgacgggcggggttatgcggcaaactcagcactgagcgtgggcttccaggcg 10440 tttattggccaggaatggcgattgagccacccgcatggtttatcgtctcccattatcatg 10500 gatgccacggtcgatcagcaaaatggttatcgcttcgtgtacagcctgccgctctcgccg 10560 accagattgttaattgaagacacgcactatatcgataatgcgacattagatcctgaacgc 10620 gcgcggcaaaatatttgcgactatgccgcgcaacagggttggcagcttcagacattgctg 10680 cgtgaagaacagggcgccttaccca 10705 3-82 Sequences 3-82-1 SequenceNumber[ID] 82 3-82-2 MolecularType DNA 3-82-3 Length 1773 source1...173 3-82-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylosinustrichosporum NonEnglishQualifierValue 3-82-5 Residues atggccagaaaaatgaccggagcggaaatggtcgtcgaagccctgaaggatcagggcgtc 60 gagattatcttcggctatcccggcggcgccgtgcttccgatctatgacgcgctcttccac 120 caggagaaggtgcagcacattctcgtgcgccacgagcagggcgccgcccatgcggccgag 180 ggctatgcgcgctcctccggcaaggtcggcgtgctgctggtcacctccggacccggcgcc 240 accaacaccatcaccggcctcaccgatgcgctgatggactccattcccgtggtctgcatc 300 accggccaggtgccgacgcatctcatcggctcggacgcctttcaagagtgcgatacggtc 360 ggcatcacccgtcactgcaccaagcataattatctggtgaagagcgtcgacgatctgccg 420 cgcattctgcacgaggccttctatgtcgcctcgagcgggcggccgggccctgtggtcatc 480 gacatccccaaggatgtgcaattcgccagcggaacctataccggcccgcgcaacgtccat 540 cacaagacctatcagcccaagctcgagggcgacacggagtctatccgccgcgccgtgaag 600 atgatggccgccgccaagcggccgatcttctacaccggcggcggcgtcatcaattccggt 660 cccgcggcctcgacgctgctgcgcgagctggtgtcgctgaccggctttccgatcacctcg 720 accttgatgggcctcggcgcctatccgggctccggccccaattggctcggcatgctcggc 780 atgcacggcaccttcgaggccaataatgcgatgcatgattgcgatctgatgatcgccgtc 840 ggcgcgcgtttcgacgatcgcatcaccggacggctcgacgccttctcgcccggctcgaag 900 aagatccacatcgatatcgatcgctcctcgatcaataagaatgtgaagatcgatctgccg 960 atcgtcggcgactgcggccatgtgctggagagtctggtgcgcgtctggcgctccgaggcg 1020 atgcacgccgagaagcagccgctcgacggctggtggaagacgatcgaccattggcgcgag 1080 cgcaagtcgctcgccttccgcaattcggacaaggtgatcaagccgcaatacgccgtgcag 1140 cggctctatgcgctcaccaaggatcgcgatccctacatcacgacggaagtcggccagcat 1200 cagatgtgggccgcgcagcattatcatttcgacgagcccaatcgctggatgacttccggc 1260 gggctcggcaccatgggctatggtctgccggcggcgatcggagcgcagctcgcgcatccg 1320 aaatcgctggtcgtcgacatcgccggcgaggcctcgatcctgatgaacattcaggagatg 1380 tcgacggcgatccaatatcggctgccggtgaaggtgttcatcctcaacaatgaatatatg 1440 ggcatggtgcgccagtggcaggagctgctgcacggcgggcgctactcgcactcctattcg 1500 gaggcgctgcccgatttcgtgaagctcgccgaagccttcgggggcaagggcatccgctgc 1560 tcggacccggcggagctcgatagcgcgattctcgagatgatcgactatgacgggccggtg 1620 atcttcgattgtctcgtcgagaaaaacgagaattgcttcccgatgatcccgtcgggcaag 1680 gcgcataacgacatgctgctcgccgatctcggcgacgacgccggcgtcgagctcggctcg 1740 atcatcgacgagaagggcaagatgctggtgtga 1773 3-83 Sequences 3-83-1 SequenceNumber[ID] 83 3-83-2 MolecularType DNA 3-83-3 Length 1056 source1...1056 3-83-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylosinustrichosporum NonEnglishQualifierValue 3-83-5 Residues atgtccaccaaagcctatgccgttgcgtccgccgaggcgctcttcggcccgctcgcgatc 60 gagcgccgcgcgctcgggcccgaggatgtagagatcgacatcctctattgcggcgtctgc 120 cattccgatctgcacacggcgcgcagtgaatggccgggcacgcgctacccatgcgtcccg 180 ggccacgagattgtcggccgcgtcaccgctgtcggcgcgaaggtgacgaaattttcggtc 240 ggcgatctcgccgccgtcggctgcatggtcgacagctgccggcgatgcttgtcctgcgac 300 gacgggctcgaacaatattgcgagcacggtttcaccgccacctataacggcccgatctac 360 ggctcgggcgagaacacctttggcggctattcggagaaaatcgtcgtcgacgcgcatttc 420 gtgctggcgatccaccattctgagacgcagcttgccggagtcgcgccgctgctctgcgcc 480 ggcatcaccacttggtcgccgctcaagcattggggtgtcggcccgggaaaatcggtcggc 540 atcgtcggcatcggcgggctcggccatatgggggtcaagctcgcccatgcgctcggcgcc 600 catgtcgtcgccttcaccacctcgccgtcaaagcgcgacgcggccctcgcgctcggcgcc 660 gacgaggtcgtcgtctccacagatcctgccgctatggcggcgcgggcgggaagcctcgac 720 ttcattctcgatacggtcgccgtcgcccatgacctcgacgcttatgtgaatctgttgaag 780 cgcgatggcgctctggtgctcgtcggcgtgccggcgacgccgcatccctcgccatcggcg 840 ggcgggttgatcttcaagcggcgccaggtcgccggctcgctgatcggcggcgtaaaggag 900 acgcaggagatgctcgacttctgcgccgagcgcggcattgtcgcggacatagagacgatc 960 gccatgcagcagatcgagaccgcctatgcgcgcatgctgaagaatgatgtgaaataccgc 1020 ttcgtcatcgacatggcgacgctgaaggcggcgtga 1056 3-84 Sequences 3-84-1 SequenceNumber[ID] 84 3-84-2 MolecularType DNA 3-84-3 Length 1029 source1...1029 3-84-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-84-5 Residues atgaaagcttgggtgatcgaccgaatcggcccgctggactcgtcgcgaactctgctacgc 60 gccaccgacctcccggtgccggagcccggccctggcgaaatcctgctgcaggtggcggtt 120 tgcggcgtctgccacaccgaaatcgacgagatcgagggccgcaccgcgccgccgcgcctg 180 ccggtcgtgcccggacaccaagcggtcggtcggatcgcggctctcggctccggcgtggcg 240 gaattcgctttgggcgaccgcgtcggcgtggcctggatcttttctgcctgcggagaatgc 300 gaattctgccggtcgggacgggagaacctctgtttcgcattctgtgccaccgggcgcgat 360 gtcgacggcggctacgcccagtacatgaccgtcccggcggcctttgctttccgcattccg 420 gagggattcaccgatgccgaagcggcgccgcttctgtgcgccggcgccatcggttaccgt 480 tcgctcaatctcagcgggctgaaaaacggccagccgctggggctcaccgggttcggggct 540 tccgcccatctggtgctgatgatggcccggtaccggtttcccgattcggaagtctatgtc 600 tttgcgcgtcatcccgaggagcgcgcgttcgcgcctcagctgggcgcggtctgggccggc 660 gacaccgcggacattgctcccgccccgctggccgccatcatcgacacgacgccggcgtgg 720 aagccggtggtcgcagcgctcgccaacctcgctcccggtggccggctggtcgttaatgcg 780 atccgcaaggcgccggacgatcgcgcctgtctcgccgaactcgactatgcccggcacttg 840 tggatggaacgggaaatcaagtcggtcgccaacgtggcgcgcgatgacgtggccgggttc 900 ctggcgctggcggcggaaatgggcatccgtcccgagacggaggagtacccgttcgaggat 960 gccgaccgggcgctgctcgacctcaagcaacgccggattcgcggggcgaaggtgttgcgg 1020 gtgacttga 1029 3-85 Sequences 3-85-1 SequenceNumber[ID] 85 3-85-2 MolecularType DNA 3-85-3 Length 1068 source1...1068 3-85-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-85-5 Residues atgcctacagccaaagcctatgccgctttttccgcagactcggcgctggcgccgttcgtc 60 ctgcagcggcgcgacccactgccccaggacatccgcatcggaatcctgtactgcggtgtc 120 tgccattccgacctgcaccaggcacgcaatgagtggaatgcgaccacatatccttgtgtg 180 ccaggccatgagatcgtcggcaaggtccttgaagtcggccgcagcgtgacgaagttcaag 240 cccggcgacacggtcgcggtgggctgcatggtggattcctgccggacctgcccgaactgc 300 gtggacgccctggaacagcactgcgagcacggccccgtcttcacctacaacagccccgat 360 ccgcacggcggcggcatgaccttcggtggctatgccgagagcatcgtggtcgacgaggcc 420 ttcgtgctgcggataccggacggactggacctcgcggccgccgccccgctgttgtgcgcc 480 gggattaccacctattcgcccctgcggcactggaaagtgggggcgggtcagcgggtcggg 540 gtcgtcggtctgggtggactgggacacatggcgctcaagttcgcgcataccttcggcgcc 600 gaaacggtgctgttcacgacgacgccggacaaggcggaggatgcccgtcggctgggagcg 660 gacgaggtcgtcgtgtcgagggatcccgaggccatggcgcggcaggccggccggttcgat 720 ttcatcctcgacaccgtctcggcgccccatgacatcgatgcctatctgaacctgctgagg 780 cgggacggcacgctgaccctggtcggcgtacctccgcaaggggtacaggtcatgcccttc 840 agcctgatcggcgggcgccggcgactggctggttcattgatcggcggcatccgggaaacc 900 caggagatgctggatttctgcggcgaacacggcatcgtctgcgacatcgagctgattccg 960 atccaaggaatcaacgacgccttcgagcgcatgctcaaaagcgacgtgaaataccgtttc 1020 gtgatcgacatggcgacgctgaacggggagtcgtccggagggcgatga 1068 3-86 Sequences 3-86-1 SequenceNumber[ID] 86 3-86-2 MolecularType DNA 3-86-3 Length 1647 source1...1647 3-86-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Lactococcuslactis NonEnglishQualifierValue 3-86-5 Residues atgtacaccgtgggcgactatctgctggaccggctgcatgaactgggcatcgaggaaatc 60 ttcggcgtccccggcgactataacctgcagttcctggaccagatcatcagccgcaaggac 120 atgaagtgggtgggcaacgccaacgagctgaacgcctcgtacatggcggacggctatgcc 180 cggaccaagaaggccgcggccttcctgaccaccttcggcgtcggcgaactgagcgccgtg 240 aacggcctggcgggctcgtacgccgagaacctgccggtcgtggaaatcgtcggctccccc 300 accagcaaggtgcagaacgagggcaagttcgtccaccataccctggccgacggcgacttc 360 aagcacttcatgaagatgcatgaaccggtgaccgcggcccgcaccctgctgaccgccgag 420 aacgcgaccgtcgaaatcgaccgcgtgctgagcgcgctgctgaaggagcggaagccggtc 480 tatatcaacctgcccgtcgacgtggcggccgcgaaggccgagaagccgtccctgcccctg 540 aagaaggaaaaccccacctcgaacacctccgaccaggagatcctgaacaagatccaggaa 600 agcctgaagaacgccaagaagccgatcgtgatcaccggccacgagatcatctcgttcggc 660 ctggaaaacaccgtcacccagttcatctccaagaccaagctgccgatcaccaccctgaac 720 ttcggcaagagctcggtggacgagaccctgccctcgttcctgggcatctacaacggcaag 780 ctgtccgaaccgaacctgaaggagttcgtggaaagcgcggacttcatcctgatgctgggc 840 gtcaagctgaccgactccagcaccggcgccttcacccaccatctgaacgagaacaagatg 900 atctcgctgaacatcgacgagggcaagatcttcaacgaatccatccagaacttcgacttc 960 gaaagcctgatctcgtccctgctggacctgtccggcatcgagtacaagggcaagtatatc 1020 gacaagaagcaggaagacttcgtcccgagcaacgcgctgctgtcgcaggaccgcctgtgg 1080 caggccgtggagaacctgacccagagcaacgagaccatcgtcgcggaacagggcacctcg 1140 ttcttcggcgccagctcgatcttcctgaagccgaagtcgcacttcatcggccagcccctg 1200 tggggctccatcggctacaccttccccgccgcgctgggctcgcagatcgcggacaaggaa 1260 tcccggcatctgctgttcatcggcgacggcagcctgcagctgaccgtgcaggagctgggc 1320 ctggccatccgcgaaaagatcaacccgatctgcttcatcatcaacaacgacggctatacc 1380 gtcgagcgggaaatccacggcccgaaccagtcgtacaacgacatccccatgtggaactat 1440 tccaagctgccggagagcttcggcgccaccgaggaacgcgtcgtgtccaagatcgtccgg 1500 accgagaacgagttcgtcagcgtgatgaaggaagcccaggcggaccccaaccggatgtac 1560 tggatcgagctggtgctggcgaaggaagacgccccgaaggtcctgaagaagatgggcaag 1620 ctgttcgccgaacagaacaagagctga 1647 3-87 Sequences 3-87-1 SequenceNumber[ID] 87 3-87-2 MolecularType DNA 3-87-3 Length 1692 source1...1692 3-87-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Saccharomycescerevisiae NonEnglishQualifierValue 3-87-5 Residues atgtcggaaatcaccctgggcaagtacctgttcgagcggctgaagcaggtcaacgtcaac 60 accatcttcggcctgcccggcgacttcaacctgagcctgctggacaagatctacgaggtc 120 gacggcctgcgctgggccggcaacgcgaacgaactgaacgccgcgtacgccgcggacggc 180 tatgcccggatcaagggcctgtcggtcctggtgaccaccttcggcgtgggcgagctgtcg 240 gccctgaacggcatcgccggctcctacgcggaacacgtcggcgtgctgcatgtcgtgggc 300 gtcccgagcatctcggcccaggcgaagcagctgctgctgcaccataccctgggcaacggc 360 gacttcaccgtgttccaccgcatgtccgccaacatcagcgagaccacctcgatgatcacc 420 gacatcgccaccgcgccgagcgaaatcgaccgcctgatccggaccaccttcatcacccag 480 cggccgtcgtacctgggcctgcccgccaacctggtcgacctgaaggtgccgggcagcctg 540 ctggagaagcccatcgacctgtcgctgaagccgaacgaccccgaggccgaaaaggaagtc 600 atcgacaccgtgctggaactgatccagaacagcaagaacctgatcgacctgtccgacgcc 660 tgcgcgagccgccacaacgtgaagaaggagacccagaagctgatcgacctgacccagttc 720 ccggccttcgtcacccccctgggcaagggctccatcgacgagcagcatccgcggtacggc 780 ggcgtctatgtgggcaccctgagcaagcaggacgtcaagcaggccgtggaaagcgcggac 840 ctgatcctgtcggtgggcgccctgctgtccgacttcaacaccggctccttcagctactcg 900 tataagaccaagaacgtcgtggagttccattcggactacgtcaaggtgaagaacgcgacc 960 ttcctgggcgtccagatgaagttcgccctgcagaacctgctgaaggtgatcccggacgtc 1020 gtgaagggctataagtccgtcccggtgcccaccaagacccccgccaacaagggcgtcccg 1080 gcgtcgacccccctgaagcaggaatggctgtggaacgagctgtccaagttcctgcaggaa 1140 ggcgacgtgatcatctcggagaccggcacctccgcgttcggcatcaaccagaccatcttc 1200 ccgaaggacgcctacggcatcagccaggtcctgtggggctcgatcggcttcaccaccggc 1260 gccaccctgggcgccgcgttcgccgcggaggaaatcgacccgaacaagcgcgtcatcctg 1320 ttcatcggcgacggctccctgcagctgaccgtgcaggaaatcagcaccatgatccggtgg 1380 ggcctgaagccctacctgttcgtgctgaacaacgacggctataccatcgagaagctgatc 1440 cacggcccgcatgcggaatacaacgagatccagacctgggaccacctggccctgctgccc 1500 gccttcggcgcgaagaagtatgaaaaccataagatcgccaccaccggcgagtgggacgcg 1560 ctgaccaccgactccgagttccagaagaacagcgtcatccgcctgatcgagctgaagctg 1620 ccggtgttcgacgcccccgaaagcctgatcaagcaggcgcagctgaccgccgcgaccaac 1680 gccaagcagtga 1692 3-88 Sequences 3-88-1 SequenceNumber[ID] 88 3-88-2 MoleculeType DNA 3-88-3 Length 1908 source1...1908 3-88-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Saccharomycescerevisiae NonEnglishQualifierValue 3-88-5 Residues atggcccccgtcaccatcgagaagttcgtcaaccaggaagagcggcatctggtgtccaac 60 cggagcgcgaccatcccgttcggcgagtacatcttcaagcgcctgctgagcatcgacacc 120 aagtcggtgttcggcgtgccgggcgacttcaacctgagcctgctggagtacctgtatagc 180 ccctcggtcgaatcggccggcctgcgctgggtgggcacctgcaacgaactgaacgccgcg 240 tacgccgcggacggctactcccggtatagcaacaagatcggctgcctgatcaccacctat 300 ggcgtcggcgaactgtcggcgctgaacggcatcgcgggctccttcgccgagaacgtgaag 360 gtcctgcacatcgtgggcgtcgccaagtcgatcgactcccgcagctcgaacttctcggac 420 cggaacctgcaccatctggtcccgcagctgcatgactccaacttcaagggccccaaccac 480 aaggtgtaccatgacatggtgaaggaccgcgtcgcgtgctccgtggcctatctggaggac 540 atcgaaaccgcctgcgaccaggtggacaacgtcatccgggacatctacaagtatagcaag 600 ccgggttacatcttcgtccccgcggacttcgccgacatgtccgtgacctgcgacaacctg 660 gtgaacgtcccgcgcatcagccagcaggactgcatcgtgtacccctccgaaaaccagctg 720 agcgacatcatcaacaagatcacctcgtggatctactccagcaagaccccggccatcctg 780 ggcgacgtcctgaccgaccggtatggcgtgagcaacttcctgaacaagctgatctgcaag 840 accggcatctggaacttctcgaccgtcatgggcaagtcggtgatcgacgaatccaacccg 900 acctacatgggccagtataacggcaaggaaggcctgaagcaggtctacgagcacttcgaa 960 ctgtgcgacctggtcctgcatttcggcgtggacatcaacgagatcaacaacggccactac 1020 accttcacctataagccgaacgcgaagatcatccagttccatcccaactacatccgcctg 1080 gtggacacccggcagggcaacgaacagatgttcaagggcatcaacttcgccccgatcctg 1140 aaggagctgtataagcgcatcgacgtcagcaagctgtcgctgcagtacgacagcaacgtg 1200 acccagtataccaacgagaccatgcggctggaagaccccaccaacggccagtcgtccatc 1260 atcacccaggtccacctgcagaagaccatgccgaagttcctgaaccccggcgacgtcgtg 1320 gtctgcgagaccggctccttccagttcagcgtgcgcgacttcgcgttcccgagccagctg 1380 aagtacatctcgcagggcttcttcctgtccatcggcatggccctgcccgccgcgctgggc 1440 gtcggcatcgcgatgcaggaccactcgaacgcccatatcaacggcggcaacgtgaaggaa 1500 gactacaagccgcggctgatcctgttcgaaggcgacggcgccgcgcagatgaccatccag 1560 gagctgtccaccatcctgaagtgcaacatcccgctggaagtcatcatctggaacaacaac 1620 ggctacaccatcgagcgcgccatcatgggccccacccggagctataacgacgtgatgtcg 1680 tggaagtggaccaagctgttcgaagcgttcggcgacttcgacggcaagtacaccaactcc 1740 accctgatccagtgcccgagcaagctggccctgaagctggaggaactgaagaactcgaac 1800 aagcgctccggcatcgagctgctggaagtcaagctgggcgagctggacttccccgaacag 1860 ctgaagtgcatggtggaggccgcggccctgaagcggaacaagaagtga 1908 3-89 Sequences 3-89-1 SequenceNumber[ID] 89 3-89-2 MoleculeType DNA 3-89-3 Length 1047 source1...1047 FeaturesLocation/ mol_type=otherDNA 3-89-4-1 Qualifiers organism=Saccharomycescerevisiae NonEnglishQualifierValue 3-89-5 Residues atgagcatccccgagacccagaaggccatcatcttctacgagagcaacggcaagctggaa 60 cataaggacatcccggtgcccaagcccaagccgaacgaactgctgatcaacgtgaagtac 120 agcggcgtctgccacaccgacctgcacgcgtggcatggcgactggccgctgcccaccaag 180 ctgcccctggtgggcggccatgaaggcgccggcgtcgtggtcggcatgggcgagaacgtc 240 aagggctggaagatcggcgactacgcgggcatcaagtggctgaacggcagctgcatggcc 300 tgcgagtattgcgaactgggcaacgaatcgaactgcccgcacgcggacctgtccggctac 360 acccatgacggcagcttccaggagtatgccaccgcggacgccgtgcaggccgcgcacatc 420 ccgcagggcaccgacctggcggaggtggcccccatcctgtgcgccggcatcaccgtctac 480 aaggcgctgaagagcgccaacctgcgcgcgggccattgggccgcgatctcgggcgccgcc 540 ggtggcctgggctccctggccgtgcagtacgcgaaggcgatgggctaccgcgtcctgggc 600 atcgacggcggtccgggcaaggaagagctgttcacctccctgggcggcgaagtgttcatc 660 gacttcaccaaggagaaggacatcgtcagcgccgtggtcaaggcgaccaacggcggcgcc 720 cacggcatcatcaacgtgtcggtctccgaagccgcgatcgaggcgtcgacccgctactgc 780 cgggccaacggcaccgtggtcctggtgggcctgcccgcgggcgccaagtgcagctcggac 840 gtcttcaaccatgtggtcaagagcatctcgatcgtgggctcgtatgtcggcaaccgcgcc 900 gacacccgcgaggccctggacttcttcgcccgtggcctggtcaagtccccgatcaaggtg 960 gtcggcctgtccagcctgcccgagatctacgaaaagatggagaagggccagatcgccggc 1020 cgctatgtggtcgacacctccaagtga 1047 3-90 Sequences 3-90-1 SequenceNumber[ID] 90 3-90-2 MoleculeType DNA 3-90-3 Length 1692 source1...1692 3-90-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Saccharomycescerevisiae NonEnglishQualifierValue 3-90-5 Residues atgagcgagatcaccctgggcaagtacctgttcgagcggctgaagcaggtcaacgtcaac 60 accgtcttcggcctgcccggcgacttcaacctgagcctgctggacaagatctacgaggtc 120 gaaggcatgcgctgggcgggcaacgccaacgagctgaacgccgcgtacgccgcggacggc 180 tatgcccggatcaagggcatgtcgtgcatcatcaccaccttcggcgtgggcgagctgtcc 240 gccctgaacggcatcgcgggcagctacgccgaacacgtcggcgtgctgcatgtcgtgggc 300 gtcccgagcatctcggcccaggcgaagcagctgctgctgcaccataccctgggcaacggc 360 gacttcaccgtgttccaccgcatgtccgcgaacatcagcgagaccaccgccatgatcacc 420 gacatcgccaccgcgccggccgaaatcgaccgctgcatccggaccacctacgtcacccag 480 cggcccgtgtatctgggcctgccggccaacctggtcgacctgaacgtgcccgcgaagctg 540 ctgcagaccccgatcgacatgtcgctgaagcccaacgacgccgagtccgaaaaggaagtc 600 atcgacaccatcctggcgctggtcaaggacgccaagaacccggtgatcctggcggacgcc 660 tgctgctcccgccacgacgtcaaggccgagaccaagaagctgatcgacctgacccagttc 720 cccgccttcgtgaccccgatgggcaagggctccatcgacgaacagcatccgcggtacggc 780 ggcgtctatgtgggcaccctgagcaagcccgaagtcaaggaagccgtggaaagcgccgac 840 ctgatcctgtcggtcggcgccctgctgtccgacttcaacaccggctccttcagctactcg 900 tataagaccaagaacatcgtggagttccacagcgaccacatgaagatccgcaacgccacc 960 ttccccggcgtccagatgaagttcgtgctgcagaagctgctgaccaccatcgccgacgcc 1020 gcgaagggctacaagccggtcgcggtgcccgcccggaccccggcgaacgccgcggtcccc 1080 gcctcgaccccgctgaagcaggaatggatgtggaaccagctgggcaacttcctgcaggaa 1140 ggcgacgtcgtgatcgcggaaaccggcacctccgccttcggcatcaaccagaccaccttc 1200 ccgaacaacacctacggcatcagccaggtgctgtggggctcgatcggcttcaccaccggc 1260 gccaccctgggcgccgcgttcgccgcggaggaaatcgacccgaagaagcgcgtcatcctg 1320 ttcatcggcgacggcagcctgcagctgaccgtgcaggaaatctcgaccatgatccggtgg 1380 ggcctgaagccctacctgttcgtcctgaacaacgacggctataccatcgagaagctgatc 1440 cacggcccgaaggcccagtacaacgaaatccagggctgggaccatctgtcgctgctgccc 1500 accttcggcgccaaggactatgagacccatcgcgtggcgaccaccggcgaatgggacaag 1560 ctgacccaggacaagtcgttcaacgacaactccaagatccggatgatcgagatcatgctg 1620 cccgtcttcgacgcgccgcagaacctggtggaacaggccaagctgaccgccgcgaccaac 1680 gcgaagcagtga 1692 3-91 Sequences 3-91-1 SequenceNumber[ID] 91 3-91-2 MoleculeType DNA 3-91-3 Length 3147 source1...3147 3-91-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Saccharomycescerevisiae NonEnglishQualifierValue 3-91-5 Residues atgaagtcggaatacaccatcggccgctatctgctggaccgcctgagcgagctgggcatc 60 cgccacatcttcggcgtccccggcgactacaacctgtcgttcctggactacatcatggag 120 tataagggcatcgactgggtcggcaactgcaacgaactgaaacgccgctacgccgcggac 180 ggctatgcccgcatcaacggcatcggcgcgatcctgaccaccttcggcgtcggcgagctg 240 tccgccatcaacgccatcgcgggcgcctacgcggaacaggtgccggtcgtgaagatcacc 300 ggcatccccaccgccaaggtccgcgacaacggcctgtatgtgcaccataccctgggcgac 360 ggccgcttcgaccacttcttcgagatgttccgggaagtcaccgtggccgaggcgctgctg 420 agcgaggaaaacgccgcgcaggaaatcgaccgcgtgctgatctcgtgctggcgccagaag 480 cggccggtcctgatcaacctgcccatcgacgtgtacgacaagccgatcaacaagccgctg 540 aagcccctgctggactataccatcagctcgaacaaggaagccgcgtgcgagttcgtcacc 600 gagatcgtgccgatcatcaaccgcgccaagaagcccgtcatcctggcggactacggcgtg 660 taccggtatcaggtccagcacgtgctgaagaacctggcggagaagaccggcttcccggtc 720 gccaccctgtcgatgggcaagggcgtgttcaacgaagcccatccgcagttcatcggcgtc 780 tacaacggcgacgtgtccagcccctatctgcgccagcgggtcgacgaggccgactgcatc 840 atctcggtcggcgtgaagctgaccgactccaccaccggcggcttctcccacggcttcagc 900 aagcgcaacgtgatccatatcgacccgttctccatcaaggccaagggcaagaagtacgcg 960 cccatcaccatgaaggacgccctgaccgaactgacctcgaagatcgagcaccggaacttc 1020 gaagacctggacatcaagccgtacaagtccgacaaccagaagtatttcgcgaaggagaag 1080 cccatcacccagaagcgcttcttcgaacggatcgcccatttcatcaaggagaaggacgtc 1140 ctgctggcggaacagggcacctgcttcttcggcgccagcaccatccagctgccgaaggac 1200 gcgaccttcatcggccagcccctgtggggctccatcggctacaccctgccggccctgctg 1260 ggcagccagctggcggaccagaagcgtcgcaacatcctgctgatcggcgacggcgccttc 1320 cagatgaccgcgcaggagatctcgaccatgctgcgcctgcagatcaagccgatcatcttc 1380 ctgatcaacaacgacggctacaccatcgagcgcgccatccacggccgggaacaggtgtac 1440 aacaacatccagatgtggcggtatcataacgtcccgaaggtgctgggccccaaggaatgc 1500 agcctgaccttcaaggtccagtcggagaccgaactggagaaggccctgctggtcgccgac 1560 aaggactgcgagcacctgatcttcatcgaagtcgtgatggaccgctacgacaagccggag 1620 cccctggaacgcctgtccaagcggttcgccaaccagaacaacggctatgcgcggatcaac 1680 ggcatcggcgccattttaaccaccttcggcgtgggcgagctgatcgcgatcaacgcgatc 1740 gccggcgcctacgcggagcaggtgccggtggtcaaaattaccggcatccccaccgcgaag 1800 gtgcgggacaacggcctgtacgtccatcacaccctgggcgacggccggttcgaccatttc 1860 ttcgaaatgttccgggaggtgaccgtcgccgaggcgctgctgtcggaagagaacgcggcc 1920 caggagatcgaccgcgtcctgatcagctgctggcggcagaagcgccccgtgctgatcaac 1980 ctgccgatcgacgtctatgacaagcccatcaacaagcccctgaagccgctgctggactac 2040 accatctcgtccaacaaggaagccgcctgcgagttcgtcaccgaaatcgtccccatcatc 2100 aaccgcgcgaagaagccggtgatcctggccgactatggcgtctatcggtatcaggtgcag 2160 catgtcctgaagaacctggccgaaaagaccggcttccccgtggccaccctgagcatgggc 2220 aagggcgtcttcaacgaggcgcacccccagttcatcggcgtgtataacggcgacgtgagc 2280 tcgccgtacctgcggcagcgcgtggacgaagccgactgcatcatcagcgtcggcgtcaag 2340 ctgaccgactcgaccaccggcggcttctcgcacggcttctcgaagcggaacgtcatccac 2400 atcgacccgttctcgatcaaggcgaagggcaagaagtatgccccgatcaccatgaaggac 2460 gcgctgaccgaactgaccagcaagatcgaacatcgcaacttcgaggacctggacatcaag 2520 ccctacaagtcggacaaccagaagtacttcgccaaggaaaagccgattactcagaagcgc 2580 ttcttcgagcgcatcgcgcacttcatcaaggaaaaggacgtcctgctggccgagcaaggc 2640 acctgcttcttcggtgcgtcgaccatccagctgcccaaggacgccaccttcatcggccag 2700 ccgctgtggggctcgatcggctataccctgcccgcgctgctgggctcccagctggccgat 2760 caaaaacgtcgcaatattttactgatcggcgacggcgcgttccagatgaccgcccaggag 2820 atcagcaccatgctgcggctgcagatcaagcccattatcttcctgattaacaacgacggc 2880 tataccatcgaacgggcgatccacggccgcgagcaggtctataataatattcaaatgtgg 2940 cggtatcataatgtgcccaaggtcctgggcccgaaggaatgctcgctgaccttcaaggtg 3000 cagagcgaaaccgagctggaaaaggccctgctggtcgccgataaggactgcgaacatctg 3060 atcttcatcgaggtggtcatggaccggtatgacaagcccgaacccctggaacggctgagc 3120 aagcgcttcgcgaaccagaacaactga 3147 3-92 Sequences 3-92-1 SequenceNumber[ID] 92 3-92-2 MoleculeType DNA 3-92-3 Length 35 3-92-4-1 FeaturesLocation/ misc_feature1...35 Qualifiers note=JPS00082 NonEnglishQualifierValue source1...35 3-92-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-92-5 Residues tgcaaggtacactgtcagaacgcagaagcggtctg 35 3-93 Sequences 3-93-1 SequenceNumber[ID] 93 3-93-2 MoleculeType DNA 3-93-3 Length 28 3-93-4-1 FeaturesLocation/ misc_feature1...28 Qualifiers note=JPS00031 NonEnglishQualifierValue source1...28 3-93-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-93-5 Residues ggtttattcctccttatttaatcgatac 28 3-94 Sequences 3-94-1 SequenceNumber[ID] 94 3-94-2 MoleculeType DNA 3-94-3 Length 34 3-94-4-1 FeaturesLocation/ misc_feature1...34 Qualifiers note=JPS00032 NonEnglishQualifierValue source1...34 mol_type=otherDNA 3-94-4-2 FeaturesLocation/ organism=syntheticconstruct Qualifiers NonEnglishQualifierValue 3-94-5 Residues aaggaggaataaaccatgggcacggttgagcctg 34 3-95 Sequences 3-95-1 SequenceNumber[ID] 95 3-95-2 MoleculeType DNA 3-95-3 Length 33 3-95-4-1 FeaturesLocation/ misc_feature1...33 Qualifiers note=GMV257 NonEnglishQualifierValue source1...33 3-95-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-95-5 Residues cacatcctgtctagatcagccctcgcccttgac 33 3-96 Sequences 3-96-1 SequenceNumber[ID] 96 3-96-2 MoleculeType DNA 3-96-3 Length 58 3-96-4-1 FeaturesLocation/ misc_feature1...58 Qualifiers note=JPS00118) NonEnglishQualifierValue source1...58 3-96-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-96-5 Residues tctagacaggatgtgtcacacaggaaaccatgtcttatcc 58 3-97 Sequences 3-97-1 SequenceNumber[ID] 97 3-97-2 MoleculeType DNA 3-97-3 Length 41 3-97-4-1 FeaturesLocation/ misc_feature1...41 Qualifiers note=JPS00119 NonEnglishQualifierValue source1...41 3-97-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-97-5 Residues acagtgtaccttgcactagtctgaaaattctttgtcgtag 41 3-98 Sequences 3-98-1 SequenceNumber[ID] 98 3-98-2 MoleculeType DNA 3-98-3 Length 70 3-98-4-1 FeaturesLocation/ misc_feature1...70 Qualifiers note=ESG00087 NonEnglishQualifierValue source1...70 3-98-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-98-5 Residues gtgttggttgctcgctagcattgtaccaagggctgagctagctataaagtcagcgccctg 60 caccattatg 70 3-99 Sequences 3-99-1 SequenceNumber[ID] 99 3-99-2 MoleculeType DNA 3-99-3 Length 46 3-99-4-1 FeaturesLocation/ misc_feature1...46 Qualifiers note=GMV251 NonEnglishQualifierValue source1...46 3-99-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-99-5 Residues gagcaaccaacacttaaagaggagaaaatgggcacggttgagcctg 46 3-100 Sequences 3-100-1 SequenceNumber[ID] 100 3-100-2 MoleculeType DNA 3-100-3 Length 149 3-100-4-1 FeaturesLocation/ misc_feature1...149 Qualifiers note=IDTgBlocksynthesizedrnpB NonEnglishQualifierValue source1...149 3-100-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-100-5 Residues gctagcactagtgatcacgtgcttaagccggcttatcggtcagtttcacctgatttacgt 60 aaaaacccgcttcggcgggtttttgcttttggaggggcagaaagatgaatgactgtccac 120 gacgctatacccaaaagaaaaccggtacc 149 3-101 Sequences 3-101-1 SequenceNumber[ID] 101 3-101-2 MoleculeType DNA 3-101-3 Length 35 3-101-4-1 FeaturesLocation/ misc_feature1...35 Qualifiers note=JPS00161 NonEnglishQualifierValue source1...35 3-101-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-101-5 Residues gcctgatacagattattgtaggtggaccagttggt 35 3-102 Sequences 3-102-1 SequenceNumber[ID] 102 3-102-2 MoleculeType DNA 3-102-3 Length 34 3-102-4-1 FeaturesLocation/ misc_feature1...34 Qualifiers note=JPS00162 NonEnglishQualifierValue source1...34 3-102-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-102-5 Residues ggtttattcctccttgatttgtcctactcaggag 34 3-103 Sequences 3-103-1 SequenceNumber[ID] 103 3-103-2 MoleculeType DNA 3-103-3 Length 34 3-103-4-1 FeaturesLocation/ misc_feature1...34 Qualifiers note=JPS00163 NonEnglishQualifierValue source1...34 3-103-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-103-5 Residues aaggaggaataaaccgctagcactagtgatcacg 34 3-104 Sequences 3-104-1 SequenceNumber[ID] 104 3-104-2 MoleculeType DNA 3-104-3 Length 34 3-104-4-1 FeaturesLocation/ misc_feature1...34 Qualifiers note=JPS00164 NonEnglishQualifierValue source1...34 3-104-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-104-5 Residues taatctgtatcaggcggtaccggttttcttttgg 34 3-105 Sequences 3-105-1 SequenceNumber[ID] 105 3-105-2 MoleculeType DNA 3-105-3 Length 34 3-105-4-1 FeaturesLocation/ misc_feature1...34 Qualifiers note=JPS00172 NonEnglishQualifierValue source1...34 3-105-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-105-5 Residues atcagactaagccttgtgcttaagccggcttatc 34 3-106 Sequences 3-106-1 SequenceNumber[ID] 106 3-106-2 MoleculeType DNA 3-106-3 Length 51 3-106-4-1 FeaturesLocation/ misc_feature1...51 Qualifiers note=JPS00173 NonEnglishQualifierValue source1...51 3-106-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-106-5 Residues aaggaggaataaaccgctagcactagtgatcacttgacggctagctcagtc 51 3-107 Sequences 3-107-1 SequenceNumber[ID] 107 3-107-2 MoleculeType DNA 3-107-3 Length 33 3-107-4-1 FeaturesLocation/ misc_feature1...33 Qualifiers note=JPS00174 NonEnglishQualifierValue source1...33 3-107-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-107-5 Residues tgaacaggtctgacttcagtgctgcgccgaggc 33 3-108 Sequences 3-108-1 SequenceNumber[ID] 108 3-108-2 MoleculeType DNA 3-108-3 Length 51 3-108-4-1 FeaturesLocation/ misc_feature1...51 Qualifiers note=JPS00176 NonEnglishQualifierValue source1...51 3-108-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-108-5 Residues agtcagacctgttcattaaagaggagaaaatgcagatttactacgacaaag 51 3-109 Sequences 3-109-1 SequenceNumber[ID] 109 3-109-2 MoleculeType DNA 3-109-3 Length 34 3-109-4-1 FeaturesLocation/ misc_feature1...34 Qualifiers note=JPS00177 NonEnglishQualifierValue source1...34 3-109-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-109-5 Residues aagtgttggttgctctcagttcttgctcgtgtcc 34 3-110 Sequences 3-110-1 SequenceNumber[ID] 110 3-110-2 MoleculeType DNA 3-110-3 Length 45 3-110-4-1 FeaturesLocation/ misc_feature1...45 Qualifiers note=JPS00157 NonEnglishQualifierValue source1...45 3-110-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-110-5 Residues gagcaaccaacacttaaagaggagaaaatgaccgacaagcacccc 45 3-111 Sequences 3-111-1 SequenceNumber[ID] 111 3-111-2 MoleculeType DNA 3-111-3 Length 33 3-111-4-1 FeaturesLocation/ misc_feature1...33 Qualifiers note=JPS00178 NonEnglishQualifierValue source1...33 3-111-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-111-5 Residues aaggcttagtctgattcagaggccgtcgtcggt 33 3-112 Sequences 3-112-1 SequenceNumber[ID] 112 3-112-2 MoleculeType DNA 3-112-3 Length 430 3-112-4-1 FeaturesLocation/ misc_feature1...430 Qualifiers note=IDTgBlocksynthesized Me-AM1PmxaF NonEnglishQualifierValue source1...430 3-112-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-112-5 Residues atggtgcagggcgcttcccgcttggtcgggccgcttcgcgagggcccgttgacgacaacg 60 gtgcgatgggtcccggccccggtcaagacgatgccaatacgttgcgacactacgccttgg 120 cacttttagaattgccttatcgtcctgataagaaatgtccgaccagctaaagacatcgcg 180 tccaatcaaagcctagaaaatataggcgaagggacgctaataagtctttcataagaccgc 240 gcaaatctaaaaatatccttagattcacgatgcggcacttcggatgacttccgagcgagc 300 ctggaacctcagaaaaacgtctgagagataccgcgaggccgaaaggcgaggcggttcagc 360 gaggagacgcaggatgagcaggtttgtgacatcagtctcggccttggcggctagcgagca 420 accaacactt 430 3-113 Sequences 3-113-1 SequenceNumber[ID] 113 3-113-2 MoleculeType DNA 3-113-3 Length 34 3-113-4-1 FeaturesLocation/ misc_feature1...34 Qualifiers note=JPS00169 NonEnglishQualifierValue source1...34 3-113-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-113-5 Residues agcgccctgcaccattatgttccggatctgcatc 34 3-114 Sequences 3-114-1 SequenceNumber[ID] 114 3-114-2 MoleculeType DNA 3-114-3 Length 46 3-114-4-1 FeaturesLocation/ misc_feature1...46 Qualifiers note=GMV00251 NonEnglishQualifierValue source1...46 3-114-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-114-5 Residues gagcaaccaacacttaaagaggagaaaatgggcacggttgagcctg 46 3-115 Sequences 3-115-1 SequenceNumber[ID] 115 3-115-2 MoleculeType DNA 3-115-3 Length 32 3-115-4-1 FeaturesLocation/ misc_feature1...32 Qualifiers note=(PS00170 NonEnglishQualifierValue source1...32 3-115-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-115-5 Residues atggtgcagggcgcttcccgcttggtcgggcc 32 3-116 Sequences 3-116-1 SequenceNumber[ID] 116 3-116-2 MoleculeType DNA 3-116-3 Length 19 3-116-4-1 FeaturesLocation/ misc_feature1...19 Qualifiers note=JPS00171 NonEnglishQualifierValue source1...19 3-116-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-116-5 Residues aagtgttggttgctcgcta 19 3-117 Sequences 3-117-1 SequenceNumber[ID] 117 3-117-2 MoleculeType DNA 3-117-3 Length 68 3-117-4-1 FeaturesLocation/ misc_feature1...68 Qualifiers note=JPS00153 NonEnglishQualifierValue source1...68 3-117-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-117-5 Residues accactcaacgatcagctagcactgtacctaggactgagctagccgtcaagtcagcgccc 60 tgcaccat 68 3-118 Sequences 3-118-1 SequenceNumber[ID] 118 3-118-2 MoleculeType DNA 3-118-3 Length 48 3-118-4-1 FeaturesLocation/ misc_feature1...48 Qualifiers note=JPS00151 NonEnglishQualifierValue source1...48 3-118-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-118-5 Residues tgatcgttgagtggtttaaagaggagaaaatgcgtgaaacgatacctc 48 3-119 Sequences 3-119-1 SequenceNumber[ID] 119 3-119-2 MoleculeType DNA 3-119-3 Length 33 3-119-4-1 FeaturesLocation/ misc_feature1...33 Qualifiers note=JPS00154 NonEnglishQualifierValue source1...33 3-119-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-119-5 Residues aagtgttggttgctctcagtgctgcgccgaggc 33 3-120 Sequences 3-120-1 SequenceNumber[ID] 120 3-120-2 MoleculeType DNA 3-120-3 Length 33 3-120-4-1 FeaturesLocation/ misc_feature1...33 Qualifiers note=JPS00183 NonEnglishQualifierValue source1...33 3-120-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-120-5 Residues tgagctagctataaagtgatcactagtgctagc 33 3-121 Sequences 3-121-1 SequenceNumber[ID] 121 3-121-2 MoleculeType DNA 3-121-3 Length 55 3-121-4-1 FeaturesLocation/ misc_feature1...55 Qualifiers note=JPS00185 NonEnglishQualifierValue source1...55 3-121-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-121-5 Residues ttatagctagctcagcccttggtacaatgctagctgatcgttgagtggtttaaag 55 3-122 Sequences 3-122-1 SequenceNumber[ID] 122 3-122-2 MoleculeType DNA 3-122-3 Length 35 3-122-4-1 FeaturesLocation/ misc_feature1...35 Qualifiers note=J23100 NonEnglishQualifierValue source1...35 3-122-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-122-5 Residues ttgacggctagctcagtcctaggtacagtgctagc 35 3-123 Sequences 3-123-1 SequenceNumber[ID] 123 3-123-2 MoleculeType DNA 3-123-3 Length 35 3-123-4-1 FeaturesLocation/ misc_feature1...35 Qualifiers note=J23100hybrid NonEnglishQualifierValue source1..35 3-123-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-123-5 Residues ttgacggctagctcagcccttggtacaatgctagc 35 3-124 Sequences 3-124-1 SequenceNumber[ID] 124 3-124-2 MoleculeType DNA 3-124-3 Length 35 3-124-4-1 FeaturesLocation/ misc_feature1...35 Qualifiers note=J23115 NonEnglishQualifierValue source1...35 3-124-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-124-5 Residues tttatagctagctcagcccttggtacaatgctagc 35 3-125 Sequences 3-125-1 SequenceNumber[ID] 125 3-125-2 MoleculeType DNA 3-125-3 Length 22 3-125-4-1 FeaturesLocation/ misc_feature1...22 Qualifiers note=GMV00233 NonEnglishQualifierValue source1...22 3-125-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue Residues 3-125-5 tgccagctgcattaatgaatcg 22 3-126 Sequences 3-126-1 SequenceNumber[ID] 126 3-126-2 MoleculeType DNA 3-126-3 Length 45 3-126-4-1 FeaturesLocation/ misc_feature1...45 Qualifiers note=GMV00235 NonEnglishQualifierValue source1...45 3-126-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-126-5 Residues ttaatgcagctggcagccagcgcttcgttaatacagatgtaggtg 45 3-127 Sequences 3-127-1 SequenceNumber[ID] 127 3-127-2 MoleculeType DNA 3-127-3 Length 52 3-127-4-1 FeaturesLocation/ misc_feature1...52 Qualifiers note=GMV00433 NonEnglishQualifierValue source1...52 3-127-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-127-5 Residues gatcgttgagtggtgaacttaaagaggagaaaatgggcacggttgagcctgg 52 3-128 Sequences 3-128-1 SequenceNumber[ID] 128 3-128-2 MoleculeType DNA 3-128-3 Length 19 3-128-4-1 FeaturesLocation/ misc_feature1...19 Qualifiers note=GMV00434 NonEnglishQualifierValue source1...19 3-128-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-128-5 Residues tgcattcgattcctgtttg 19 3-129 Sequences 3-129-1 SequenceNumber[ID] 129 3-129-2 MoleculeType DNA 3-129-3 Length 19 3-129-4-1 FeaturesLocation/ misc_feature1...19 Qualifiers note=GMV00435 NonEnglishQualifierValue source1...19 3-129-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-129-5 Residues caggaatcgaatgcaaccg 19 3-130 Sequences 3-130-1 SequenceNumber[ID] 130 3-130-2 MoleculeType DNA 3-130-3 Length 49 3-130-4-1 FeaturesLocation/ misc_feature1...49 Qualifiers note=GMV00436 NonEnglishQualifierValue source1...49 3-130-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-130-5 Residues gagcaaccaacactcacacaggaaaccatgcatattacatacgatctgc 49 3-131 Sequences 3-131-1 SequenceNumber[ID] 131 3-131-2 MoleculeType DNA 3-131-3 Length 39 3-131-4-1 FeaturesLocation/ misc_feature1...39 Qualifiers note=GMV00437 NonEnglishQualifierValue source1...39 3-131-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-131-5 Residues gttcaccactcaacgatcttaagcgtcaacgaaaccggt 39 3-132 Sequences 3-132-1 SequenceNumber[ID] 132 3-132-2 MoleculeType DNA 3-132-3 Length 69 3-132-4-1 FeaturesLocation/ misc_feature1...69 Qualifiers note=GMV00438 NonEnglishQualifierValue source1...69 3-132-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-132-5 Residues tgaacaggtctgactgctagcattgtaccaagggctgagctagctataaagatttgtcct 60 actcaggag 69 3-133 Sequences 3-133-1 SequenceNumber[ID] 133 3-133-2 MoleculeType DNA 3-133-3 Length 53 3-133-4-1 FeaturesLocation/ misc_feature1...53 Qualifiers note=GMV00439 NonEnglishQualifierValue source1...53 3-133-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-133-5 Residues agtcagacctgttcattaaagaggagaaaatgagcggaaaaaccctttacgac 53 3-134 Sequences 3-134-1 SequenceNumber[ID] 134 3-134-2 MoleculeType DNA 3-134-3 Length 42 3-134-4-1 FeaturesLocation/ misc_feature1...42 Qualifiers note=GMV00440 NonEnglishQualifierValue source1...42 3-134-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-134-5 Residues aaggcttagtctgattcagacggcgcgcaaggcggcgacgat 42 3-135 Sequences 3-135-1 SequenceNumber[ID] 135 3-135-2 MoleculeType DNA 3-135-3 Length 49 3-135-4-1 FeaturesLocation/ misc_feature1...49 Qualifiers note=GMV00441 NonEnglishQualifierValue source1...49 3-135-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-135-5 Residues atcagactaagcctttcacacaggaaaccatgcacgacagactgatcat 49 3-136 Sequences 3-136-1 SequenceNumber[ID] 136 3-136-2 MoleculeType DNA 3-136-3 Length 35 3-136-4-1 FeaturesLocation/ misc_feature1...35 Qualifiers note=GMV00442 NonEnglishQualifierValue source1...35 3-136-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-136-5 Residues ggtttattcctcctttcacacatccccgacttgcg 35 3-137 Sequences 3-137-1 SequenceNumber[ID] 137 3-137-2 MoleculeType DNA 3-137-3 Length 49 3-137-4-1 FeaturesLocation/ misc_feature1...49 Qualifiers note=ESG00084 NonEnglishQualifierValue source1...49 3-137-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-137-5 Residues gagacagaacgaagtgtgaccagaacgcagaagcggtctgataaaacag 49 3-138 Sequences 3-138-1 SequenceNumber[ID] 138 3-138-2 MoleculeType DNA 3-138-3 Length 70 3-138-4-1 FeaturesLocation/ misc_feature1...70 Qualifiers note=ESG00088 NonEnglishQualifierValue source1...70 3-138-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-138-5 Residues gtgttggttgctcgctagcactgtacctaggactgagctagccgtcaagtcagcgccctg 60 caccattatg 70 3-139 Sequences 3-139-1 SequenceNumber[ID] 139 3-139-2 MoleculeType DNA 3-139-3 Length 10705 3-139-4-1 FeaturesLocation/ misc_feature1...10705 Qualifiers note=pMZT37 NonEnglishQualifierValue source1...10705 3-139-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-139-5 Residues tcaccctgtcgggcaatgccgaggcattctggcagcagcgccccctggcctgtagtggat 60 tacgtgccggtctgttccatcctaccaccggctattcactgccgctggcggttgccgtgg 120 ccgaccgcctgagcgcacttgatgtctttacgtcggcctcaattcaccaggctattaggc 180 attttgcccgcgagcgctggcagcagcagcgctttttccgcatgctgaatcgcatgctgt 240 ttttagccggacccgccgattcacgctggcgggttatgcagcgtttttatggtttacctg 300 aagatttaattgcccgtttttatgcgggaaaactcacgctgaccgatcggctacgtattc 360 tgagcggcaagccgcctgttccggtattagcagcattgcaagccattatgacgactcatc 420 gttaagagacagaacgaagtgtgaccagaacgcagaagcggtctgataaaacagaatttg 480 cctggcggcagtagcgcggtggtcccacctgaccccatgccgaactcagaagtgaaacgc 540 cgtagcgccgatggtagtgtggggtctccccatgcgagagtagggaactgccaggcatca 600 aataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttctttgtcggt 660 gaacgctctcctgagtaggacaaatcttgtaggtggaccagttggtgattttgaactttt 720 gctttgccacggaacggtctgcgttgtcgggaagatgcgtgatctgatccttcaactcag 780 caaaagttcgatttattcaacaaagccgccgtcccgtcaagtcagcgtaatgctctgcca 840 gtgttacaaccaattaaccaattctgattagaaaaactcatcgagcatcaaatgaaactg 900 caatttattcatatcaggattatcaataccatatttttgaaaaagccgtttctgtaatga 960 aggagaaaactcaccgaggcagttccataggatggcaagatcctggtatcggtctgcgat 1020 tccgactcgtccaacatcaatacaacctattaatttcccctcgtcaaaaataaggttatc 1080 aagtgagaaatcaccatgagtgacgactgaatccggtgagaatggcaaaagcttatgcat 1140 ttctttccagacttgttcaacaggccagccattacgctcgtcatcaaaatcactcgcatc 1200 aaccaaaccgttattcattcgtgattgcgcctgagcgagacgaaatacgcgatcgctgtt 1260 aaaaggacaattacaaacaggaatcgaatgcaaccggcgcaggaacactgccagcgcatc 1320 aacaatattttcacctgaatcaggatattcttctaatacctggaatgctgttttcccggg 1380 gatcgcagtggtgagtaaccatgcatcatcaggagtacggataaaatgcttgatggtcgg 1440 aagaggcataaattccgtcagccagtttagtctgaccatctcatctgtaacatcattggc 1500 aacgctacctttgccatgtttcagaaacaactctggcgcatcgggcttcccatacaatcg 1560 atagattgtcgcacctgattgcccgacattatcgcgagcccatttatacccatataaatc 1620 agcatccatgttggaatttaatcgcggcctcgagcaagacgtttcccgttgaatatggct 1680 cataacaccccttgtattactgtttatgtaagcagacagttttattgttcatgatgatat 1740 atttttatcttgtgcaatgtaacatcagagattttgagacacaacgtggctttccccccc 1800 ccccctgcaggtccgacacggggatggatggcgttcccgatcatggtcctgcttgcttcg 1860 ggtggcatcggaatgccggcgctgcaagcaatgttgtccaggcaggtggatgaggaacgt 1920 caggggcagctgcaaggctcactggcggcgctcaccagcctgacctcgatcgtcggaccc 1980 ctcctcttcacggcgatctatgcggcttctataacaacgtggaacgggtgggcatggatt 2040 gcaggcgctgccctctacttgctctgcctgccggcgctgcgtcgcgggctttggagcggc 2100 gcagggcaacgagccgatcgctgatcgtggaaacgataggcctatgccatgcgggtcaag 2160 gcgacttccggcaagctatacgcgccctagaattgtcaattttaatcctctgtttatcgg 2220 cagttcgtagagcgcgccgtgcgtcccgagcgatactgagcgaagcaagtgcgtcgagca 2280 gtgcccgcttgttcctgaaatgccagtaaagcgctggctgctgaacccccagccggaact 2340 gaccccacaaggccctagcgtttgcaatgcaccaggtcatcattgacccaggcgtgttcc 2400 accaggccgctgcctcgcaacgcttcgcaggcttcgccgacctgctcgcgccacttcttc 2460 acgcgggtggaatccgatccgcacatgaggcggaaggtttccagcttgagcgggtacggc 2520 tcccggtgcgagctgaaatagtcgaacatcctgcgggccgtcggcgacagcttgcggtac 2580 ttctcccatatgaatttcgtgtagtggtcgccagcaaacagcacgacgatttcctcgtcg 2640 atcaggacctggcaacgggacgttttcttgccacggtccaggacgcggaagcggtgcagc 2700 agcgacaccgattccaggtgcccaacgcggtcggacgtgaagcccatcgccgtcgcctgt 2760 aggcgcgacaggcattcctcggccttcgtgtaataccggccattgatcgaccagcccagg 2820 tcctggcaaagctcgtagaacgtgaaggtgatcggctcgccgataggggtgcgcttcgcg 2880 tactccaacacctgctgccacaccagttcgtcatcgtcggcccgcagctcgacgccggtg 2940 taggtgatcttcacgtccttgttgacgtggaaaatgaccttgttttgcagcgcctcgcgc 3000 gggattttcttgttgcgcgtggtgaacagggcagagcgggccgtgtcgtttggcatcgct 3060 cgcatcgtgtccggccacggcgcaatatcgaacaaggaaagctgcatttccttgatctgc 3120 tgcttcgtgtgtttcagcaacgcggcctgcttggcctcgctgacctgttttgccaggtcc 3180 tcgccggcggtttttcgcttcttggtcgtcatagttcctcgcgtgtcgatggtcatcgac 3240 ttcgccaaacctgccgcctcctgttcgagacgacgcgaacgctccacggcggccgatggc 3300 gcgggcagggcagggggagccagttgcacgctgtcgcgctcgatcttggccgtagcttgc 3360 tggaccatcgagccgacggactggaaggtttcgcggggcgcacgcatgacggtgcggctt 3420 gcgatggtttcggcatcctcggcggaaaaccccgcgtcgatcagttcttgcctgtatgcc 3480 ttccggtcaaacgtccgattcattcaccctccttgcgggattgccccgactcacgccggg 3540 gcaatgtgcccttattcctgatttgacccgcctggtgccttggtgtccagataatccacc 3600 ttatcggcaatgaagtcggtcccgtagaccgtctggccgtccttctcgtacttggtattc 3660 cgaatcttgccctgcacgaataccagctccgcgaagtcgctcttcttgatggagcgcatg 3720 gggacgtgcttggcaatcacgcgcaccccccggccgttttagcggctaaaaaagtcatgg 3780 ctctgccctcgggcggaccacgcccatcatgaccttgccaagctcgtcctgcttctcttc 3840 gatcttcgccagcagggcgaggatcgtggcatcaccgaaccgcgccgtgcgcgggtcgtc 3900 ggtgagccagagtttcagcaggccgcccaggcggcccaggtcgccattgatgcgggccag 3960 ctcgcggacgtgctcatagtccacgacgcccgtgattttgtagccctggccgacggccag 4020 caggtaggcctacaggctcatgccggccgccgccgccttttcctcaatcgctcttcgttc 4080 gtctggaaggcagtacaccttgataggtgggctgcccttcctggttggctgcagagcagg 4140 agccatccgcttgccctcatctgttacgccggcggtagccggccagcctcgcagagcagg 4200 attcccgttgagcaccgccaggtgcgaataagggacagtgaagaaggaacacccgctcgc 4260 gggtgggcctacttcacctatcctgcccggctgacgccgttggatacaccaaggaaagtc 4320 tacacgaaccctttggcaaaatcctgtatatcgtgcgaaaaaggatggatataccgaaaa 4380 aatcgctataatgaccccgaagcagggttatgcagcggaaaagatccgtcgaccctttcc 4440 gacgctcaccgggctggttgccctcgccgctgggctggcggacgttgacacttgaggggc 4500 cgcgccagaaacgccgtcgaagccgtgtgcgagacaccgcggccgccggcgttgtggata 4560 cctcgcggaaaacttggccctcactgacagatgaggggcggacgttgacacttgaggggc 4620 cgactcacccggcgcggcgttgacagatgaggggcaggctcgatttcggccggcgacgtg 4680 gagctggccagcctcgcaaatcggcgaaaacgcctgattttacgcgagtttcccacagat 4740 gatgtggacaagcctggggataagtgccctgcggtattgacacttgaggggcgcgactac 4800 tgacagatgaggggcgcgatccttgacacttgaggggcagagtgctgacagatgaggggc 4860 gcacctattgacatttgaggggctgtccacaggcagaaaatccagcatttgcaagggttt 4920 ccgcccgtttttcggccaccgctaacctgtcttttaacctgcttttaaaccaatatttat 4980 aaaccttgtttttaaccagggctgcgccctgtgcgcgtgaccgcgcacgccgaagggggg 5040 tgcccccccttctcgaaccctcccggcccgctaacgcgggcctcccatccccccaggggc 5100 tgcgcccctcggccgcgaacggcctcaccccaaaaatggcagccaagctgaccacttctg 5160 cgctcggcccttccggctggctggtttattgctgataaatctggagccggtgagcgtggg 5220 tctcgcggtatcattgcagcactggggccagatggtaagccctcccgtatcgtagttatc 5280 tacacgacggggagtcaggcaactatggatgaacgaaatagacagatcgctgagataggt 5340 gcctcactgattaagcattggtaactgtcagaccaagtttactcatatatactttagatt 5400 gatttaaaacttcatttttaatttaaaaggatctaggtgaagatcctttttgataatctc 5460 atgaccaaaatcccttaacgtgagttttcgttccactgagcgtcagaccccgtagaaaag 5520 atcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaa 5580 aaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccg 5640 aaggtaactggcttcagcagagcgcagataccaaatactgtccttctagtgtagccgtag 5700 ttaggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctg 5760 ttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacga 5820 tagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagc 5880 ttggagcgaacgacctacaccgaactgagatacctacagcgtgagctatgagaaagcgcc 5940 acgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacagga 6000 gagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggttt 6060 cgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatgg 6120 aaaaacgccagcaacgcggcccttttacggttcctggccttttgctggcctgtcgggttt 6180 atgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtga 6240 gctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcg 6300 gaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagc 6360 tggcagccagcgcttcgttaatacagatgtaggtgttccacagggtagccagcagcatcc 6420 tgcgatgcagatccggaacataatggtgcagggcgctgacttgacggctagctcagtcct 6480 aggtacagtgctagcgagcaaccaacacttaaagaggagaaaatgtatccgtttataagg 6540 acagcccgaatgacggtctgcgcaaaaaaacacgttcatctcactcgcgatgctgcggag 6600 cagttactggctgatattgatcgacgccttgatcagttattgcccgtggagggagaacgg 6660 gatgttgtgggtgccgcgatgcgtgaaggtgcgctggcaccgggaaaacgtattcgcccc 6720 atgttgctgttgctgaccgcccgcgatctgggttgcgctgtcagccatgacggattactg 6780 gatttggcctgtgcggtggaaatggtccacgcggcttcgctgatccttgacgatatgccc 6840 tgcatggacgatgcgaagctgcggcgcggacgccctaccattcattctcattacggagag 6900 catgtggcaatactggcggcggttgccttgctgagtaaagcctttggcgtaattgccgat 6960 gcagatggcctcacgccgctggcaaaaaatcgggcggtttctgaactgtcaaacgccatc 7020 ggcatgcaaggattggttcagggtcagttcaaggatctgtctgaaggggataagccgcgc 7080 agcgctgaagctattttgatgacgaatcactttaaaaccagcacgctgttttgtgcctcc 7140 atgcagatggcctcgattgttgcgaatgcctccagcgaagcgcgtgattgcctgcatcgt 7200 ttttcacttgatcttggtcaggcatttcaactgctggacgatttgaccgatggcatgacc 7260 gacaccggtaaggatagcaatcaggacgccggtaaatcgacgctggtcaatctgttaggc 7320 cctagggcggttgaagaacgtctgagacaacatcttcatcttgccagtgagcatctctct 7380 gcggcctgccaacacgggcacgccactcaacattttattcaggcctggtttgacaaaaaa 7440 ctcgctgccgtcagttaaaggtctctagacaggatgtgtcacacaggaaaccatgaaacc 7500 aactacggtaattggtgcaggcttcggtggcctggcactggcaattcgtctacaggctgc 7560 ggggatccccgtcttactgcttgaacaacgtgataaacccggcggtcgggcttatgtcta 7620 cgaggatcaggggtttacctttgatgcaggcccgacggttatcaccgatcccagtgccat 7680 tgaagaactgtttgcactggcaggaaaacagttaaaagagtatgtcgaactgctgccggt 7740 tacgccgttttaccgcctgtgttgggagtcagggaaggtctttaattacgataacgatca 7800 aacccggctcgaagcgcagattcagcagtttaatccccgcgatgtcgaaggttatcgtca 7860 gtttctggactattcacgcgcggtgtttaaagaaggctatctgaagctcggtactgtccc 7920 ttttttatcgttcagagacatgcttcgcgccgcacctcaactggcgaaactgcaggcatg 7980 gagaagcgtttacagtaaggttgccagttacatcgaagatgaacatctgcgccaggcgtt 8040 ttctttccactcgctgttggtgggcggcaatcccttcgccacctcatccatttatacgtt 8100 gatacacgcgctggagcgtgagtggggcgtctggtttccgcgtggcggcaccggcgcatt 8160 agttcaggggatgataaagctgtttcaggatctgggtggtgaagtcgtgttaaacgccag 8220 agtcagccatatggaaacgacaggaaacaagattgaagccgtgcatttagaggacggtcg 8280 caggttcctgacgcaagccgtcgcgtcaaatgcagatgtggttcatacctatcgcgacct 8340 gttaagccagcaccctgccgcggttaagcagtccaacaaactgcagactaagcgtatgag 8400 taactctctgtttgtgctctattttggtttgaatcaccatcatgatcagctcgcgcatca 8460 cacggtttgtttcggcccgcgttaccgcgaactgattgacgagatttttaatcatgatgg 8520 cctcgcagaagacttctcactttatctgcacgcgccctgtgtcacggattcgtcactggc 8580 gcctgaaggttgcggcagttactatgtgttggcgccggtgccgcatttaggcaccgcgaa 8640 cctcgactggacggttgaggggccaaaactacgcgaccgtatttttgagtaccttgagca 8700 gcattacatgcctggcttacggagtcagctggtcacgcaccagatgtttacgccgtttga 8760 ttttcgcgaccagcttaatgcctatcagggctcagccttttctgtggagcccgttcttac 8820 ccagagcgcctggtttcggccgcataaccgcgataaaaccattactaatctctacctggt 8880 cggcgcaggcacgcatcccggcgcaggcattcctggcgtcatcggctcggcaaaagcgac 8940 agcaggtttgatgctggaggatctgatttaagtgatcgttgagtggtgaacttaaagagg 9000 agaaaatgaataatccgtcgttactcaatcatgcggtcgaaacgatggcagttggctcga 9060 aaagttttgcgacagcctcaaagttatttgatgcaaaaacccggcgcagcgtactgatgc 9120 tctacgcctggtgccgccattgtgacgatgttattgacgaccagacgctgggcttccagg 9180 cccggcagcctgccttacaaacgcccgaacaacgtctgatgcaacttgagatgaaaacgc 9240 gccaggcctatgcaggatcgcagatgcacgaaccggcgtttgcggcttttcaggaagtgg 9300 ctatggctcatgatatcgccccggcttacgcgtttgatcatctggaaggcttcgccatgg 9360 atgtacgcgaagcgcaatacagccaactggacgatacgctgcgctattgctatcacgttg 9420 caggcgttgtcggcttgatgatggcgcaaatcatgggcgtacgggataacgccacgctgg 9480 accgcgcctgtgaccttgggctggcatttcagttgaccaatattgctcgcgatattgtgg 9540 acgatgcgcatgcgggccgctgttatctgccggcaagctggctggagcatgaaggtctga 9600 acaaagagaattatgcggcacctgaaaaccgtcaggcgctgagccgtatcgcccgtcgtt 9660 tggtgcaggaagcagaaccttactatttgtctgccacagcgggcctggctgggttgcccc 9720 tgcgttcggcctgggcaatcgctacggcgaagcaggtttaccggaaaataggtgtcaaag 9780 ttgaacaggccggtcagcaagcctgggatcagcggcagtcaacgaccacgcccgaaaaat 9840 taacgctgctgctggccgcctctggtcaggcccttacttcccggatgcgggctcatcctc 9900 cccgccctgcgcatctctggcagcgcccgctctaatcacgtagcaagctgacagtttaaa 9960 gaggagaaaatgggagcggctatgcaaccgcattatgatctgattctcgtgggggctgga 10020 ctcgcgaatggccttatcgccctgcgtcttcagcagcagcaacctgatatgcgtattttg 10080 cttatcgacgccgcaccccaggcgggcgggaatcatacgtggtcatttcaccacgatgat 10140 ttgactgagagccaacatcgttggatagcttcgctggtggttcatcactggcccgactat 10200 caggtacgctttcccacacgccgtcgtaagctgaacagcggctacttctgtattacttct 10260 cagcgtttcgctgaggttttacagcgacagtttggcccgcacttgtggatggataccgcg 10320 gtcgcagaggttaatgcggaatctgttcggttgaaaaagggtcaggttatcggtgcccgc 10380 gcggtgattgacgggcggggttatgcggcaaactcagcactgagcgtgggcttcccagcg 10440 tttattggccaggaatggcgattgagccacccgcatggtttatcgtctcccattatcatg 10500 gatgccacggtcgatcagcaaaatggttatcgcttcgtgtacagcctgccgctctcgccg 10560 accagattgttaattgaagacacgcactatatcgataatgcgacattagatcctgaacgc 10620 gcgcggcaaaatatttgcgactatgccgcgcaacagggttggcagcttcagacattgctg 10680 cgtgaagaacagggcgccttaccca 10705 3-140 Sequences 3-140-1 SequenceNumber[ID] 140 3-140-2 MoleculeType DNA 3-140-3 Length 411 3-140-4-1 FeaturesLocation/ misc_feature1...411 Qualifiers note=MaFAR-g1 NonEnglishQualifierValue source1...411 3-140-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-140-5 Residues gagcaaccaacacttaaagaggagaaaatggcgacccagcagcagcagaacggcgcctcg 60 gcgagcggcgtcctggaacagttgcgcgggaagcatgtcctgataaccggtaccaccggt 120 ttccttggcaaggtagtcctggaaaagctgatccgcacagtcccggacatcggcggcatc 180 cacctcctgatccggggcaacaagaggcatccggccgcccgtgaacggttcttgaacgag 240 atcgccagcagttcggtcttcgagcgtctgcgccacgacgacaacgaggccttcgaaacc 300 ttcctggaagaaagggtgcactgtataaccggagaggtcaccgagagtcgtttcggcctt 360 accccggagcgcttccgcgcgctggcgggtcaggtggacgccttcatcaat 411 3-141 Sequences 3-141-1 SequenceNumber[ID] 141 3-141-2 MoleculeType DNA 3-141-3 Length 465 3-141-4-1 FeaturesLocation/ misc_feature1...465 Qualifiers note=MaFAR-g2 NonEnglishQualifierValue 3-141-4-2 FeaturesLocation/ source1...465 Qualifiers mol_type=otherDNA organism=syntheticconstruct NonEnglishQualifierValue 3-141-5 Residues gacgccttcatcaattcggccgcctccgtcaacttccgcgaggaactggacaaggcgctg 60 aagatcaatacgctgtgcctggagaatgtcgcggcccttgctgaactcaacagtgcgatg 120 gcggtcatccaggtttcgacctgctacgttaacggcaagaatagcgggcagatcaccgaa 180 tcggtcatcaagcccgcgggggagtccatcccgcgtagcaccgatgggtactatgaaatc 240 gaagaattggtgcacctgctgcaggacaaaatcagcgatgtgaaggcccgatactccggg 300 aaggttctggaaaaaaaattggtggacctaggcatccgggaagccaataactacgggtgg 360 agcgatacatataccttcaccaagtggctgggcgaacagctcctcatgaaggccctgagc 420 ggcagatcgctgaccatcgtgcggccgtcgatcatcgagtcggca 465 3-142 Sequences 3-142-1 SequenceNumber[ID] 142 3-142-2 MoleculeType DNA 3-142-3 Length 374 3-142-4-1 FeaturesLocation/ misc_feature1...374 Qualifiers note=MaFAR-g3 NonEnglishQualifierValue source1...374 3-142-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-142-5 Residues atcatcgagtcggcattggaagagcccagcccggggtggattgaaggcgtcaaggtcgcc 60 gatgccatcatactggcctacgcgagggagaaggtatcgctctttcctggcaagcggagc 120 ggcatcatcgacgtcatcccagtggatctggtggccaattcgatcattctgtccctggcg 180 gaggcgctctccggttcgggccagcggcgtatctatcagtgctgcagcggcggctcgaac 240 cccatctccctcgggaagttcatcgactatctgatggcggaggcgaagaccaactacgcg 300 gcctacgatcagctgttctaccgccgccccaccaagccgttcgtggccgtcaaccgcaaa 360 ctcttcgacgtcgt 374 3-143 Sequences 3-143-1 SequenceNumber[ID] 143 3-143-2 MoleculeType DNA 3-143-3 Length 376 3-143-4-1 FeaturesLocation/ misc_feature1...376 Qualifiers note=MaFAR-g4 NonEnglishQualifierValue source1..376 3-143-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-143-5 Residues actcttcgacgtcgtcgtgggcggcatgcgggtcccgctctcgatcgcgggcaaagccat 60 gcgcctggcgggacaaaaccgcgaactgaaggtcctgaagaatctggatacgacccggtc 120 cctggccaccattttcgggttctacaccgctccggactacatctttcgcaatgacagcct 180 gatggccctggcctcgcgcatgggcgagctggaccgcgtgttgttccccgttgacgcccg 240 tcagatcgactggcagctgtatctgtgcaaaatccacctcggcgggctgaatcggtacgc 300 gctcaaggaacgtaagctgtactcgctccgggccgccgacactcgcaagaaggcagcctg 376 agagacagaacgaagt 3-144 Sequences 3-144-1 SequenceNumber[ID] 144 3-144-2 MoleculeType DNA 3-144-3 Length 47 3-144-4-1 FeaturesLocation/ misc_feature1...47 Qualifiers note=GMV410 NonEnglishQualifierValue source1...47 3-144-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-144-5 Residues gagcaaccaacacttaaagaggagaaaatgcgccccctgcaccccat 47 3-145 Sequences 3-145-1 SequenceNumber[ID] 145 3-145-2 MoleculeType DNA 3-145-3 Length 47 3-145-4-1 FeaturesLocation/ misc_feature1...47 Qualifiers note=GMV411 NonEnglishQualifierValue source1...47 3-145-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-145-5 Residues gagcaaccaacacttaaagaggagaaaatgcgcctgctgaccgccgt 47 3-146 Sequences 3-146-1 SequenceNumber[ID] 146 3-146-2 MoleculeType DNA 3-146-3 Length 47 3-146-4-1 FeaturesLocation/ misc_feature1...47 Qualifiers note=GMV412 NonEnglishQualifierValue source1...47 3-146-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-146-5 Residues gagcaaccaacacttaaagaggagaaaatgtccgtgatgtccccgac 47 3-147 Sequences 3-147-1 SequenceNumber[ID] 47 3-147-2 MoleculeType DNA 3-147-3 Length 47 3-147-4-1 FeaturesLocation/ misc_feature1...47 Qualifiers note=GMV413 NonEnglishQualifierValue source1...47 3-147-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-147-5 Residues gagcaaccaacacttaaagaggagaaaatgccggtcaccgactccat 47 3-148 Sequences 3-148-1 SequenceNumber[ID] 148 3-148-2 MoleculeType DNA 3-148-3 Length 47 3-148-4-1 FeaturesLocation/ misc_feature1...47 Qualifiers note=GMV414 NonEnglishQualifierValue source1...47 3-148-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-148-5 Residues gagcaaccaacacttaaagaggagaaaatggccccgaccgactccct 47 3-149 Sequences 3-149-1 SequenceNumber[ID] 149 3-149-2 MoleculeType DNA 3-149-3 Length 47 3-149-4-1 FeaturesLocation/ misc_feature1...47 Qualifiers note=GMV415 NonEnglishQualifierValue source1...47 3-149-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-149-5 Residues gagcaaccaacacttaaagaggagaaaatgcccctgccgatgtcccc 47 3-150 Sequences 3-150-1 SequenceNumber[ID] 150 3-150-2 MoleculeType DNA 3-150-3 Length 35 3-150-4-1 FeaturesLocation/ misc_feature1...35 Qualifiers note=GMV416 NonEnglishQualifierValue source1...35 3-150-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-150-5 Residues acttcgttctgtctctcagttggcggtcttgatgt 35 3-151 Sequences 3-151-1 SequenceNumber[ID] 151 3-151-2 MoleculeType DNA 3-151-3 Length 35 3-151-4-1 FeaturesLocation/ misc_feature1...35 Qualifiers note= ACTTCGTTCTGTCTCTCACGGGGCCAGCTTCTTCA NonEnglishQualifierValue source1...35 3-151-4-2 FeaturesLocation/Q mol_type=otherDNA ualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-151-5 Residues acttcgttctgtctctcacggggccagcttcttca 35 3-152 Sequences 3-152-1 SequenceNumber[ID] 152 3-152-2 MoleculeType DNA 3-152-3 Length 35 3-152-4-1 FeaturesLocation/ misc_feature1...35 Qualifiers note=GMV418 NonEnglishQualifierValue source1...35 3-152-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-152-5 Residues acttcgttctgtctctcaggtgccgctcgcggcca 35 3-153 Sequences 3-153-1 SequenceNumber[ID] 153 3-153-2 MoleculeType DNA 3-153-3 Length 35 3-153-4-1 FeaturesLocation/ misc_feature1...35 Qualifiers note=GMV419 NonEnglishQualifierValue source1...35 3-153-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-153-5 Residues acttcgttctgtctctcacagcagggccgcttcca 35 3-154 Sequences 3-154-1 SequenceNumber[ID] 154 3-154-2 MoleculeType DNA 3-154-3 Length 35 3-154-4-1 FeaturesLocation/ miscfeature1...35 Qualifiers note=GMV420 NonEnglishQualifierValue source1...35 3-154-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-154-5 Residues acttcgttctgtctctcacaggccgaccgcggttt 35 3-155 Sequences 3-155-1 SequenceNumber[ID] 155 3-155-2 MoleculeType DNA 3-155-3 Length 35 3-155-4-1 FeaturesLocation/ misc_feature1...35 Qualifiers note=GMV421 NonEnglishQualifierValue source1...35 3-155-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-155-5 Residues acttcgttctgtctctcagatgcccaccgcgcgtt 35 3-156 Sequences 3-156-1 SequenceNumber[ID] 156 3-156-2 MoleculeType DNA 3-156-3 Length 1149 source1...1149 3-156-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Saccharomycescerevisiae NonEnglishQualifierValue 3-156-5 Residues atgcgggcgctggcgtatttcaagaagggcgacatccatttcaccaacgacatcccccgg 60 ccggagatccagaccgacgacgaagtgatcatcgacgtctcctggtgcggcatctgcggc 120 agcgacctgcacgagtacctggacggccccatcttcatgccgaaggacggcgaatgccac 180 aagctgagcaacgccgcgctgcccctggcgatgggccatgagatgtcgggcatcgtctcc 240 aaagtgggcccgaaggtgaccaaggtcaaagtgggcgaccacgtcgtggtcgacgccgcg 300 agctcgtgcgccgacctgcactgctggccccattccaagttctataacagcaagccgtgc 360 gacgcctgccagcgcggctcggagaacctgtgcacccatgcgggcttcgtcggcctgggc 420 gtgatcagcggcggcttcgccgaacaggtggtcgtgtcgcagcaccatatcatcccggtc 480 cccaaggagatccccctggacgtcgccgccctggtcgagccgctgtcggtcacctggcac 540 gccgtgaagatctccggcttcaagaagggctccagcgccctggtcctgggcgcgggcccc 600 atcggcctgtgcaccatcctggtgctgaagggcatgggcgcgtcgaagatcgtcgtgtcc 660 gagatcgccgaacgtcgcatcgagatggcgaagaagctgggcgtcgaagtgttcaacccg 720 agcaagcacggccataagtcgatcgagatcctgcggggcctgaccaagtcccacgacggc 780 ttcgactacagctatgactgctcgggcatccaggtcaccttcgaaaccagcctgaaggcc 840 ctgaccttcaagggcaccgccaccaacatcgcggtctggggcccgaagcccgtgccgttc 900 cagccgatggacgtcaccctgcaggagaaggtgatgaccggctcgatcggctacgtcgtg 960 gaagacttcgaggaagtcgtgcgcgccatccataacggcgacatcgcgatggaggactgc 1020 aagcagctgatcaccggcaagcagcggatcgaggacggctgggaaaagggcttccaggag 1080 ctgatggaccacaaggaatccaacgtgaagatcctgctgaccccgaacaaccacggcgaa 1140 atgaagtga 1149 3-157 Sequences 3-157-1 SequenceNumber[ID] 157 3-157-2 MoleculeType AA 3-157-3 Length 382 source1...382 3-157-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Saccharomycescerevisiae NonEnglishQualifierValue 3-157-5 Residues MRALAYFKKGDIHFTNDIPRPEIQTDDEVIIDVSWCGICGSDLHEYLDGPIFMPKDGECH 60 KLSNAALPLAMGHEMSGIVSKVGPKVTKVKVGDHVVVDAASSCADLHCWPHSKFYNSKPC 120 DACQRGSENLCTHAGFVGLGVISGGFAEQVVVSQHHIIPVPKEIPLDVAALVEPLSVTWH 180 AVKISGFKKGSSALVLGAGPIGLCTILVLKGMGASKIVVSEIAERRIEMAKKLGVEVFNP 240 SKHGHKSIEILRGLTKSHDGFDYSYDCSGIQVTEFTSKLALTFKGTATNIAVWGPKPVPF 300 QPMDVTLQEKVMTGSIGYVVEDFEEVVRAIHNGDIAMEDCKQLITGKQRIEDGWEKGFQE 360 LMDHKESNVKILLTPNNHGEMK 382 3-158 Sequences 3-158-1 SequenceNumber[ID] 158 3-158-2 MoleculeType DNA 3-158-3 Length 47 3-158-4-1 FeaturesLocation/ misc_feature1...47 Qualifiers note=GMV00268 NonEnglishQualifierValue source1...47 3-158-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-158-5 Residues gagcaaccaacacttaaagaggagaaaatgcgggcgctggcgtattt 47 3-159 Sequences 3-159-1 SequenceNumber[ID] 159 3-159-2 MoleculeType DNA 3-159-3 Length 35 3-159-4-1 FeaturesLocation/ misc_feature1...35 Qualifiers note=GMV00271 NonEnglishQualifierValue source1...35 3-159-4-2 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=syntheticconstruct NonEnglishQualifierValue 3-159-5 Residues acttcgttctgtctctcacttcatttcgccgtggt 35 3-160 Sequences 3-160-1 SequenceNumber[ID] 160 3-160-2 MoleculeType DNA 3-160-3 Length 990 source1...990 3-160-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Escherichiacoli NonEnglishQualifierValue 3-160-5 Residues atgcatattacatacgatctgccggttgctattgatgacattattgaagcgaaacaacga 60 ctggctgggcgaatttataaaacaggcatgcctcgctccaactattttagtgaacgttgc 120 aaaggtgaaatattcctgaagtttgaaaatatgcagcgtacgggttcatttaaaattcgt 180 ggcgcatttaataaattaagttcactgaccgatgcggaaaaacgcaaaggcgtggtggcc 240 tgttctgcgggcaaccatgcgcaaggggtttccctctcctgcgcgatgctgggtatcgac 300 ggtaaagtggtgatgccaaaaggtgcgccaaaatccaaagtagcggcaacgtgcgactac 360 tccgcagaagtcgttctgcatggtgataacttcaacgacactatcgctaaagtgagcgaa 420 attgtcgaaatggaaggccgtatttttatcccaccttacgatgatccgaaagtgattgct 480 ggccagggaacgattggtctggaaattatggaagatctctatgatgtcgataacgtgatt 540 gtgccaattggtggtggcggtttaattgctggtattgcggtggcaattaaatctattaac 600 ccgaccattcgtgttattggcgtacagtctgaaaacgttcacggcatggcggcttctttc 660 cactccggagaaataaccacgcaccgaactaccggcaccctggcggatggttgtgatgtc 720 tcccgcccgggtaatttaacttacgaaatcgttcgtgaattagtcgatgacatcgtgctg 780 gtcagcgaagacgaaatcagaaacagtatgattgccttaattcagcgcaataaagtcgtc 840 accgaaggcgcaggcgctctggcatgtgctgcattattaagcggtaaattagaccaatat 900 attcaaaacagaaaaaccgtcagtattatttccggcggcaatatcgatctttctcgcgtc 960 tctcaaatcaccggtttcgttgacgcttaa 990 3-161 Sequences 3-161-1 SequenceNumber[ID] 161 3-161-2 MoleculeType AA 3-161-3 Length 329 source1...329 3-161-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Escherichiacoli NonEnglishQualifierValue 3-161-5 Residues MHITYDLPVAIDDIIEAKQRLAGRIYKTGMPRSNYFSERCKGEIFLKFENMQRTGSFKIR 60 GAFNKLSSLTDAEKRKGVVACSAGNHAQGVSLSCMALGIDGKVVMPKGAPKSKVAATCDY 120 SAEVVLHGDNFNDTIAKVSEIVEMEGRIFIPPYDDPKVIAGQGTIGLEIMEDLYDVDNVI 180 VPIGGGGLIAGIAVAIKSINPTIRVIGVQSENVHGMAASFHSGEITTHRTTGTLADGCDV 240 SRPGNLTYEIVRELVDDIVLVSEDEIRNSMIALIQRNKVVTEGAGALACAALLSGKLDQY 300 IQNRKTVSIISGGNIDLSRVSQITGFVDA 329 3-162 Sequences 3-162-1 SequenceNumber[ID] 162 3-162-2 MoleculeType AA 3-162-3 Length 590 source1...590 3-162-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism= Methylosinustrichosporium NonEnglishQualifierValue 3-162-5 Residues MARKMTGAEMVVEALKDQGVEIIFGYPGGAVLPIYDALFHQEKVQHILVRHEQGAAHAAE 60 GYARSSGKVGVLLVTSGPGAENTITGLTDALMDSIPVVCITGQVPTHLIGSDAFGECDTV 120 GITRHCTKHNYLVKSVDDLPRILHEAFYVASSGRPGPVVIDIPKDVQFASGTYTGPRNVH 180 HKTYQPKLEGDTESIRRAVKMMAAAKRPIFYTGGGVINSGPAASTLLRELVSLTGFPITS 240 TLMGLGAYPGSGPNWLGMLGMHGTFEANNAMHDCDLMIAVGARFDDRITGRLDAFSPGSK 300 KIHIDIDRSSINKNVKIDLPIVGDCGHVLESLVRVWRSEAMHAEKQPLDGWMKTIDHWRE 360 RKSLAFRNSDKVIKPQYAVQRLYALTKDRDPYITTEVGQHQMWAAQHYHFDEPNRWMTSG 420 GLGTMGYGLPAAIGAQLAHPKSLVVDIAGEASILMNIQEMSTAIQYRLPVKVFILNNEYM 480 GMVRQWQELLHGGRYSHSYSEALPDFVKLAEAFGGKGIRCSDPAELDSAILEMIDYDGPV 540 IFDCLVEKNENCFPMIPSGKAHNDMLLADLGDDAGVELGSIIDEKGKMLV 590 3-163 Sequences 3-163-1 SequenceNumber[ID] 163 3-163-2 MoleculeType AA 3-163-3 Length 351 source1...351 3-163-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism= NonEnglishQualifierValue Methylosinustrichosporium 3-163-5 Residues MSTKAYAVASAEALFGPLAIERRALGPEDVEIDILYCGVCHSDLHTARSEWPGTRYPCVP 60 GHEIVGRVTAVGAKVTKFSVGDLAAVGDMVDSCRRCLSCDDGLEQYCEHGFTATYNGPIY 120 GSGENTFGGYSEKIVVDAHFVLAIHHSETQLAGVAPLLCAGITTWSPLKHWGVGPGKSVG 180 IVGIGGLGHMGVKLAHALGAHVVAFTTSPSKRDAALALGADEVVVSTDPAAMAARAGSLD 240 FILDTVAVAHDLDAYVNLLKRDGALVLVGVPATPHPSPSAGGLIFKRRQVAGSLIGGVKE 300 TQEMLDECAERGIVADIETIAMQQIETAYARMLKNDVKYRFVIDMATKLAA 351 3-164 Sequences 3-164-1 SequenceNumber[ID] 164 3-164-2 MoleculeType AA 3-164-3 Length 342 source1...342 3-164-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-164-5 Residues MKAWVIDRIGPLDSSRTLLRATDLPVPEPGPGEILLQVAVCGVCHTEIDEIEGRTAPPRL 60 PVVPGHQAVGRIAALGSGVAEFALGDRVGVAWIFSACGECEFCRSGRENLCFAFCATGRD 120 VDGGYAQYMTVPAAFAFRIPEGFTDAEAAPLLCAGAIGYRSLNLSGLKNGQPLGLTGFGA 180 SAHLVLMMARYRFPDSEVYVFARHPEERAFALQLGAVWAGDTADIAPAPLAAIIDTTPAW 240 KPVVAALANLAPGGRLVVNAIRKAPDDRACLAELDYARHLWMEREIKSVANVARSDVAGF 300 LALAAEMGIRPETEEYPFEDADRALLDLKQRRIRGAKVLRVT 342 3-165 Sequences 3-165-1 SequenceNumber[ID] 165 3-165-2 MoleculeType AA 3-165-3 Length 355 source1...355 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-165-5 Residues MPTAKAYAAFSADSALAPFVLQRRDPLPQDIRIGILYCGVCHSDLHQARNEWNATTYPCV 60 PGHEIVGKVLEVGRSVTKFKPGDTVAVGCMVDSCRTCPNCVDALEQHCEHGPVFTYNSPC 120 PHGGGMTFGGYAESIVVDEAFVLRIPDGLDLAAAAPLLCAGITTYSPLRHWKVGAGQRVG 180 VVGLGGLGHMALKFAHTFGAETVLFTTTPDKAEDARRLGADEVVVSRDPEAMARQAGRFD 240 FILDTVSAPHDIDAYLNLLRRDGTLTLVGVPPQGVQVMPFSLIGGRRRLAGSLIGGIRET 300 QEMLDFCGEHGIVCDIELIPIQGINDAFERMLKSDVKYRFVIDMATLNGESSGGR 355 3-166 Sequences 3-166-1 SequenceNumber[ID] 166 3-166-2 MoleculeType AA 3-166-3 Length 548 source1...548 3-166-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Lactococcuslactis NonEnglishQualifierValue 3-166-5 Residues MYTVGDYLLDRLHELGIEEIFGVPGDYNLQFLDQIISRKDMKWVGNANELNASYMADGYA 60 RTKKAAAFLTTFGVGELSAVNGLAGSYAENLPVVEIVGSPTSKVQNEGKFVHHTLADGDF 120 KHFMKMHEPVTAARTLLTAENATVEIDRVLSALLKERKPVYINLPVDVAAAKAEKPSLPL 180 KKENPTSNTSDQEILNKIQESLKNAKKPIVITGHEIISFGLENTVTQFISKTKLPITTLN 240 FGKSSVDETLPSFLGIYNGKLSEPNLKEFVESADFILMLGVKLTDSSTGAFTHHLNENKM 300 ISLNIDEGKIFNESIQNFDFESLISSLLDLSGIEYKGKYIDKKQEDFVPSNALLSQDRLW 360 QAVENLTQSNETIVAEQGTSFFGASSIFLKPKSHFIGQPLWGSIGYTFPASYNDIPMWNY 420 SRHLLFIGDGSLQLTVQELGLAIREKINPICFIINNDGYTVEREIHGPNQSYNDIPMWNY 480 SKLPESFGATEERVVSKIVRTENEFVSVMKEAGADPNRMYWIELVLAKEDAPKVLKKMGK 540 LFAEQNKS 548 3-167 Sequences 3-167-1 SequenceNumber[ID] 167 3-167-2 MoleculeType AA 3-167-3 Length 563 source1...563 3-167-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Saccharomycescerevisiae NonEnglishQualifierValue 3-167-5 Residues MSEITLGKYLFERLKQVNVNTIFGLPGDFNLSLLDKIYEVDGLRWAGNANELNAAYAADG 60 YARIKGLSVLVTTFGVGELSALNGIAGSYAEHVGVLHVVGVPSISAGAKQLLLHHTLGNG 120 DFTVFHRMSANISETTSMITDIATAPSEIDRLIRTTFITQRPSYLGLPANLVDLKVPGSL 180 LEKPIDLSLKPNDPEAEKEVIDTVLELIQNSKNPVILSDACASRHNVKKETQKLIKLTQF 240 PAFVTPLGKGSIDEQHPRYGGVYVGTLSKQDVLQAVESADLILSVGALLSDFNTGSFSYS 300 YKTKNVVEFHSDYVKVKNATFLGVQMKFALQNLLKVIPDVVKGYKSVPVPTKTPANKGVP 360 ASTPLKQEWLWNELSKFLQEGDVIISETGTSAFGINQTIFPKDAYGISQVLWGSIGFTTG 420 ATLGAAFAAEEIDPNKRVILFIGDGSLQLTVQEISTMIRWGLKPYLFVLNNDGYTIEKLI 480 HGPHAEYNEIQTWDHLALLPAFGAKKYENHKIATTGEWDALTTDSEFQKNSVIRLIELKL 540 PVFDAPESLIKQAQLTAATNAKQ 563 3-168 Sequences 3-168-1 SequenceNumber[ID] 168 3-168-2 MoleculeType AA 3-168-3 Length 635 source1...635 3-168-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Saccharomycescerevisiae NonEnglishQualifierValue 3-168-5 Residues MAPVTIEKFVNQEERHLVSNRSATIPFGEYIFKRLLSIDTKSVFGVPGDFNLSLLEYLYS 60 PSVESAGLRWVGTCNELNAAYAADGYSRYSNKIGCLITTYGVGELSALNGIAGSFAENVK 120 VLHIVGVAKSIDSRSSNFSDRNLHHLVPQLHDSNFKGPNHKVYHDMVKDRVACSVAYLED 180 IETACDQVDNVIRDIYKYSKPGYIFVPADFADMSVTCDNLVNVPRISQQDCIVYPSENQL 240 SDIINKITSWIYSSKTPAILGDVLTDRYGVSNFLNKLICKTGIWNFSTVMGKSVIDESNP 300 TYMGQYNGKEGLKQVYEHFELCDLVLHFGVDINEINNGHYTFTYKPNAKIIQFHPNYIRL 360 VDTRQGNEQMFKGINFAPILKELYKRIDVSKLSLQYDSNVTQYTNETMRLEDPTNGQSSI 420 ITQVHLQKTMPKFLNPGDVVVCETGSFQFSVRDFAFPSQLKYISQGFFLSIGMALPAALG 480 VGIAMQDHSNAHINGGNVKEDYKPRLILFEGDGAAQMTIQELSTILKCNIPLEVIIWNNN 540 GYTIERAIMGPTRSYNDVMSWKWTKLFEAFGDFDGKYTNSTLIQCPSKLALKLEELKNSN 600 KRSGIELLEVKLGELDFPEQLKCMVEAAALKRNKK 635 3-169 Sequences 3-169-1 SequenceNumber[ID] 169 3-169-2 MoleculeType AA 3-169-3 Length 348 source1...348 3-169-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Saccharomycescerevisiae NonEnglishQualifierValue 3-169-5 Residues MSIPETQKAIIFYESNGKLEHKDIPVPKPKPHELLINVKYSGVCHTDLHAWHGDWPLPTK 60 LPLVGGHEGAGVVVGMGENVKGWKIGDYAGIKWLNGSCMACEYCELGNESNCPHADLSGY 120 THDGSEQEYATADAVQAAHIPQGTDLAEVAPILCAGITVYKALKSANLRAGHWAAISGAA 180 GGLGSLAVQYAKAMGYRVLGIDGGPGKEELFTSLGGEVFIDFTKEKDIVSAVVKATNGGA 240 HGIINVSVSEAAIEASTRYCRANGTVVLVGLPAGAKCSSDVFNHVVKSISIVGSYVGNRA 300 DTREALDEFARGLVKSPIKVVGLSSLPEIYEKMEKGQIAGRYVVTDSK 348 3-170 Sequences 3-170-1 SequenceNumber[ID] 170 3-170-2 MoleculeType AA 3-170-3 Length 563 source1...563 3-170-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Saccharomycescerevisiae NonEnglishQualifierValue 3-170-5 Residues MSEITLGKYLFERLKQVNVNTVFGLPGDFNLSLLDKIYEVEGMRWAGNANELNAAYAADG 60 YARIKGMSCIITTFGVGELSALNGIAGSYAEHVGVLHVVGVPSISAQAKQLLLHHTLGNG 120 DFTVFHRMSANISETTAMITDIATAPAEIDRCIRTTYVTQRPVYLGLPANLVDLNVPAKL 180 LQTPIDMSLKPNDAESEKEVIDTILALVKDAKNPVILADACCSRHDVKAETKKLIDLTQF 240 PAFVTPMGKGSIDEQHPRYGGVYVGTLSKPEVKEAVESADLILSVGALLSDFNTGSFSYS 300 YKTKNIVEFHSDHMKIRNATFPGVQMKFVLQKLLTTIADAAKGYKPVAVPARTPANAAVP 360 ASTPLKQEWMWNQLGNFLQEGDVVIAETGTSAFGINQTTFPNNTYGISQVLWGSIGFTTG 420 ATLGAAFAAEEIDPKKRVILFIGDGSLQLTVQEISTMIRWGLKPYLFVLNNDGYTIEKLI 480 HGPKAQYNEIQGWDHLSLLPTFGAKDYETHRVATTGEWDKLTQDKSFNDNSKIRMIEIML 540 PVFDAPQNLVEQAKLTAATNAKQ 563 3-171 Sequences 3-171-1 SequenceNumber[ID] 171 3-171-2 MoleculeType AA 3-171-3 Length 1048 source1...1048 3-171-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism= Clostridiumacetobutylicum NonEnglishQualifierValue 3-171-5 Residues MKSEYTIGRYLLDRLSELGIRHIFGVPGDYNLSFLDYIMEYKGIDWVGNCNELNAGYAAD 60 GYARINGIGAILTTFGVGELSAINAIAGAYAEQVPVVKITGIPTAKVRDNGLYVHHTLGD 120 GRFDHFFEMFREVTVAEALLSEENAAQEIDRVLISCWRQKRRVLINLPIDVYDKPINKPL 180 KPLLDYTISSNKEAACEFVTEIVPIINRAKKPVILADYGVYRYQVQHVLKNLAEKTGFPV 240 ATLSMGKGVFNEAHPQFIGVYNGDVSSPYLRQRVDEADCIISVGVKLTDSTTGGFSHGFS 300 KRNVIHIDPFSIKAKGKKYAPITMKDALTELTSKIEHRNFEDLDIKPYKSDNQKYFAKEK 360 PITQKRFFERIAHFIKEKDVLLAEQGTCFFGASTIQLPKDATFIGQPLWGSIGYTLPALL 420 GSQLADQKRRNILLIGDGAFQMTAQEISTMLRLQIKPIIFLINNDGYTIERAIHGREQVY 480 NNIQMWRYHNVPKVLGPKECSLTFKVQSETELEKALLVADKDCEHLIFIEVVMDRYDKPE 540 PLERLSKRFANQNNGYARINGIGAILTTFGVGELSAINAIAGAYAEQVPVVKITGIPTAK 600 VRDNGLYVHHTLGDGRFDHFFEMFREVTVAEALLSEENAAQEIDRVLISCWRQKRPVLIN 660 LPIDVYDKPINKPLKPLLDYTISSNKEAACEFVTEIVPIINRAKKPVILADYGVYRVQVQ 720 HVLKNLAEKTGFPVATLSMGKGVFNEAHPQFIGVYNGDVSSPYLRQRVDEADCIISVGVK 780 LTDSTTGGFSHGFSKRNVIHIDPFSIKAKGKKYAPITMKDALTELTSKIEHRNFEDLDIK 840 PYKSDNQKYFAKEKPITQKRFFERIAHFIKEDKVLLAEQGTCFFGASTIQLPKDATFIGQ 900 PLWGSIGYTLPALLGSQLADQKRRNILLIGDGAFQMTAQEISTMLRLQIKPIIFLINNDG 960 YTIERAIHGREQVYNNIQMWRYHNVPKVLGPKECSLTFKVQSETELEKALIVADKDCEHL 1020 IFIEVVMDRYDKPEPLERLSKRFANQNN 1048 3-172 Sequences 3-172-1 SequenceNumber[ID] 172 3-172-2 MoleculeType DNA 3-172-3 Length 645 source1...645 3-172-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-172-5 Residues atggcaagaccattgattcagctcgccctggacacgctggacatcccgcagaccctgaag 60 ctcgcaagcctcaccgcgccctatgtcgatatcttcgaaatcggcacccccagcatcaag 120 cacaacggcatcgccctggtgaaggagttcaaaaaacgcttccccaacaagctgctcctg 180 gtcgacctcaaaaccatggacgccggtgaatacgaagccacccccttcttcgccgccggc 240 gccgacatcaccaccgtcctcggcgtcgcaggactggccaccatcaagggcgtcatcaac 300 gccgccaacaagcacaacgccgaggtccaggtcgacctgatcaacgtccccgacaaggcc 360 gcctgcgcccgtgagtccgccaaggccggcgcccagatcgtcggcatccacaccggcctc 420 gacgcccaggccgccggccagacccccttcgccgacctccaggccatcgccaagctcggc 480 ctccccgtccgcatctccgtcgccggcggcatcaaggcctccaccgcccaacaggtcgtc 540 aaaaccggtgccaacatcatcgtcgtcggagccgccatctacggcgccgcctcccccgcc 600 gatgccgcgcgcgaaatctacgaacaggtcgtcgccgcttccgcc 645 3-173 Sequences 3-173-1 SequenceNumber[ID] 17 3-173-2 MoleculeType AA 3-173-3 Length 215 source1...215 3-173-4 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-173-5 Residues MARPLIQLALDTLDIPQTLKLASLTAPYVDIFEIGTPSIKHNGIALVKEFKKRFPNKLLL 60 VDLKTMDAGEYEATPFFAAGADITTVLGVAGLATIKGVINAANKNHAEVQVDLINVPDKA 120 ACARESAKAGAQIVGIHTGLDAQAAGQTPFADLQAIAKLGLPVRISVAGGIKASTAQQVV 180 KTGANIIVVGAAIYGAASPADAAREIYEQVVAASA 215 3-174 Sequences 3-174-1 SequenceNumber[ID] 174 3-174-2 MoleculeType DNA 3-174-3 Length 531 source1..531 3-174-4-1 FeaturesLocation/ mol_type=otherDNA Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-174-5 Residues atgcatcagaaactgatcatagacaaaatctccggcatcctcgccgccaccgatgccggc 60 tatgatgcaaaactgactgccatgctcgaccaggcctcccgcatcttcgtcgcgggggcc 120 ggccggtcggggctggtcgccaagttcttcgccatgcgcctcatgcacggcggctatgac 180 gtcttcgtcgtcggcgaaatcgtcacccccagcatccgcaagggcgacttgctgatcgtg 240 atctccggctccggtgaaaccgaaaccatgctcgccttcaccaaaaaagccaaggagcag 300 ggcgcctccatcgccctcatctccacccgcgacagctcctccctcggcgacctcgccgac 360 tccgtcttccgcatcggctccccagagctcttcggaaaagtcgtcggcatgcccatgggc 420 accgtcttcgagctctccaccctcctcttcctcgaggccaccatctctcacatcatccac 480 gagaaaggcatccccgaagaagaaatgagaactcgtcacgccaacctggaa 531 3-175 Sequences 3-175-1 SequenceNumber[ID] 175 3-175-2 MoleculeType AA 3-175-3 Length 177 source1..77 3-175-4-1 FeaturesLocation/ mol_type=protein Qualifiers organism=Methylococcuscapsulatus NonEnglishQualifierValue 3-175-5 Residues MHQKLIIDKISGILAATDAGYDAKLTAMLDQASRIFVAGAGRSGLVAKFFAMRLMHGGYD 60 VFVVGEIVTPSIRKGDLLIVISGSGETETMLAFTKKAKEQGASIALISTRDSSSLGDLAD 120 SVFRIGSPELFGKVVGMPMGTVFELSTLLFLEATISHIIHEKGIPEEEMRTRHANLE 177