COMPOSITIONS AND METHODS
20220001007 · 2022-01-06
Inventors
- Sarah C. GILBERT (Oxford, GB)
- Sarah SEBASTIAN (Oxford, GB)
- Marta ULASZEWSKA (Oxford, GB)
- Adrian V.S. HILL (Oxford, GB)
- Teresa LAMBE (Oxford, GB)
Cpc classification
International classification
Abstract
The invention relates to a viral vector comprising nucleic acid having a polynucleotide sequence encoding at least one epitope of the varicella-zoster virus (VZV) Gly E antigen, wherein said viral vector is an adenoviral vector. The invention also relates to uses, compositions for use in medical treatments, and methods of medical treatment.
Claims
1. A composition comprising a viral vector comprising nucleic acid having a polynucleotide sequence encoding at least one epitope of the varicella-zoster virus (VZV) Gly E antigen, wherein the viral vector is an adenoviral vector.
2. The composition of claim 1 wherein the at least one epitope comprises at least one CD4 T cell epitope and at least one CD8 T cell epitope.
3. The composition of claim 1 wherein the adenoviral vector is of human or simian origin.
4. The composition of claim 1 wherein the adenoviral vector is selected from the group consisting of ChAdOx 1 and ChAdOx 2.
5. The composition of claim 1 wherein the composition is adjuvant-free.
6. The composition of claim 1 wherein the Gly E antigen has the amino acid sequence is selected from the group consisting of SEQ ID NO: 1 and SEQ ID NO: 2.
7. The composition of claim 1 wherein the polynucleotide sequence comprises a sequence selected from the group consisting of SEQ ID NO: 3 and SEQ ID NO: 4.
8. The composition of claim 1 wherein the polynucleotide sequence further comprises the sequence of the bgh polyadenylation signal SEQ ID NO: 6.
9. The composition of claim 1 wherein the polynucleotide sequence encoding at least one epitope of the varicella-zoster virus (VZV) Gly E antigen is operably connected to the long CMV promoter, wherein the long CMV promoter has the nucleotide sequence of SEQ ID NO: 7.
10. The composition of claim 1 wherein the viral vector sequence is as in ECACC accession number 12052403 (ChAdOx1).
11. The composition of claim 1 wherein the viral vector comprises the sequence of SEQ ID NO: 5 (ChAdOx2).
12. The composition of claim 1 wherein the composition is configured such that administration of a single dose of the composition to a mammalian subject induces protective immunity in the subject.
13. The composition of claim 1 wherein the composition is configured to induce an immune response against VZV in a subject by administration of the composition to the subject.
14. (canceled)
15. (canceled)
16. The composition of claim 1, wherein the composition is configured such that annual administration to a subject is capable of inducing an immune response against VZV in the subject.
17. The composition of claim 1, wherein the composition is configured for use in preventing VZV infection in a subject by administration of the composition to the subject.
18. The composition of claim 1, wherein the composition is configured for use in prevention of shingles in a subject by administration of the composition to the subject.
19. (canceled)
20. (canceled)
21. (canceled)
22. (canceled)
23. (canceled)
24. (canceled)
25. (canceled)
26. A method of inducing an immune response against varicella-zoster virus (VZV) in a mammalian subject, the method comprising the steps of administering at least one dose of the composition of claim 1 to the subject.
27. A method of preventing shingles in a mammalian subject, the method comprising the steps of administering at least one dose of the composition of claim 1 to the subject.
28. (canceled)
29. (canceled)
30. The method of claim 26 wherein the composition is administered annually.
31. The method of claim 26 wherein the composition is administered by a route of administration selected from the group consisting of subcutaneous, intradermal and intramuscular administration.
32. (canceled)
33. (canceled)
34. The composition of claim 1 wherein the composition is configured for use in treatment or prevention of chickenpox in a subject.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
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EXAMPLES
Example 1—Viral Vectored Vaccines Against VZV
[0275] We have generated viral vectored vaccines toward VZV.
[0276] Our data suggest that the vaccine of the invention outperforms a currently licensed prior art Zoster vaccine as assessed for CMI pre-clinically (
[0277] The higher CMI routinely achieved with viral vectored vaccines, when compared to other vaccine modalities, is likely to translate to higher efficacy, while advantageously only a single shot of viral vectored vaccine may be required for efficacy in contrast to repeated administration of known protein-in-adjuvant vaccines.
[0278] We refer to
[0279] Groups of Balb/c mice (n=5) were vaccinated intramuscularly with 1×10.sup.7 IU of ChAdOx1-VZVgpE or 1×10.sup.7 IU of ChAdOx2-VZVgpE or 1.3×10.sup.3 pfu Zostax.
[0280] Splenocytes were collected 2 weeks after final vaccination and the cellular immune response against peptides spanning the whole glycoprotein-E were measured by ELISpot analysis.
[0281] Responses post ChAdOx1-VZV-gE were significantly higher than those post Zostavax.
[0282] This test is in young mice. Age was approx. >8 weeks.
[0283] It is noted that the prior art Zostavax vaccine can replicate in humans and without wishing to be bound by theory partial immunogenicity may be argued to have come from this. However, it has been demonstrated that non-replicating Zostavax vaccine is comparable in terms of measured immunogenicity to replicating Zostavax in man and can induce a similar immune response.
[0284] In any case, this is a fair test because none of the vaccines used replicate in mice.
[0285] Thus it is demonstrated that the invention outperforms prior art Zostavax.
[0286] We refer to
[0287] 8 wk.sup.+ old or aged ex-breeder female Balb/c mice were vaccinated intramuscularly with 1.00E+07 iu of ChAdOX1-VZV-gE Mice were culled approx. 2 weeks later and spleen ELISpot performed with peptides spanning the entire VZV gE insert.
[0288] Responses post ChAdOx1-VZV-gE were not significantly different.
[0289] This test is in aged mice.
[0290] We refer to
[0291] 8 wk.sup.+ old female Balb/c mice were vaccinated intramuscularly with ChAdOX1-VZV-gE—group 1. 1.00E+07 iu ChAdOX1-VZV-gE then four weeks later boosted with 1.00E+07 iu ChAdOX1-VZV-gE ChAdOX2-VZV-gE—group 2. 1.00E+07 iu ChAdOX2-VZV-gE then four weeks later boosted with 1.00E+07 iu ChAdOX2-VZV-gE.
[0292] Zostavax group 3. 1.29E+03 VZV Zostavax then four weeks later boosted with 1.29E+03 VZV Zostavax. Sera was taken at the indicated timepoints and assayed for anti-VZV-gpE specific antibodies.
[0293] The inventors note that Kruskal-Wallis analysis shows with Dunn's multiple comparisons test significant difference in response between group 2 and 3, but not between group 1 and group 3 at 2 wk post-boost. This may represent a further advantage of this particular embodiment where the vector is ChAdOx1 i.e. the inventors would not have expected to see antibody response comparable to Zostavax (as evidenced by group 2—ChAdOx2) but group 1 (ChAdOx1 embodiment) generates a surprisingly good antibody response as well as good T cell responses.
[0294] The inventors note that there is no significant difference in responses in
[0295] We refer to
[0296] lane 1. 1.00E+07 iu ChAdOX1-VZV-gE then one week later boosted with 1.00E+07 iu ChAdOX1-VZV-gE. (filled box)
[0297] lane 1. 1.00E+07 iu ChAdOX1-VZV-gE then one week later boosted with 1.00E+07 iu ChAdOX2-VZV-gE. (filled circle)
[0298] lane 2. 1.00E+07 iu ChAdOX1-VZV-gE then four weeks later boosted with 1.00E+07 iu ChAdOX1-VZV-gE (filled box)
[0299] lane 2. 1.00E+07 iu ChAdOX1-VZV-gE then four weeks later boosted with 1.00E+07 iu ChAdOX2-VZV-gE. (filled circle)
[0300] lane 3 (no Boost). 1.00E+07 iu ChAdOX1-VZV-gE (open circle)
[0301] Mice were culled approx. 2 weeks later and spleen ELISpot performed with peptides spanning the entire VZV gE insert.
[0302] Thus, lane 3 (no boost) represents a “one-shot” scheme; lanes 2 and 3 ‘filled boxes’ represent ‘homologous prime-boost’ schemes; lanes 2 and 3 ‘filled circle’ represent ‘heterologous prime-boost’ schemes. It could be argued that a heterologous second shot does not show augmentation—however, augmentation might be expected to show at a later time point—this is considered to be due to a response-curve effect.
[0303] Responses post ChAdOx1-VZV-gE were not significantly different across in young or aged animals for single-administration applications.
[0304] A preferred interval between prime and boost (in prime-boost applications; overall single-administration embodiments are preferred) is 4 weeks; when prime and boost are both Ad vectors, the interval may be for example 2, 4, 6 or 8 weeks.
[0305] Mouse Model System
[0306] Regarding the mouse model system for testing these vaccines, it should be noted that a 25 non-replicating zoster virus can give the same response as a replicating zoster virus in humans. Therefore, the mouse data presented herein do indeed represent a fair comparison since although the prior art Zostavax™ does induce a limited infection in humans which is important to boosting the immune response, neither the adenoviral vector constructs of the invention nor the Zostavax prior art comparator can replicate 30 in mice, and therefore the data provided in the application comparing those to formulations in mice are indeed fair and indicative of the superior properties of the vectors according to the invention.
Example 2: Vectors of the Invention Express Efficiently
[0307] We present western blot analysis of viral vector expression of VZVgpE. Subconfluent HEK293T (ChAdOx 1) were infected with viruses with at the indicated MOI. Cells were harvested 18 h later, lysed and protein supernatant lysate run on a Biorad 4-12% gradient gel and probed with a 1:1000 dilution of abeam 52549 VZV in 0.05% PBST and expression detected with ECL reagent.
[0308] Results are shown in
[0316] Thus it is demonstrated that the compositions of the invention produce expression of the antigen in human cells.
Example 3: Immunogenicity of Viral Vectors Encoding Varicella Zoster Virus Glycoprotein E
[0317] We demonstrate cellular immunogenicity after one-shot vaccination against VZV. We refer to
[0318] We refer to
[0319] These ELISpot data show that a cellular immune response, as demonstrated by the T cell response, is induced according to the invention. The response is evident at 2 weeks. The response is induced by a single administration. The response is induced by a single dose. The response is a sustained response as shown by the data at the 16 week timepoints.
[0320] We refer to
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[0325] Thus overall these FACS sorted experiments show that triple secreting CD4+ T cells (which are very good as without wishing to be bound by theory they are considered the most protective) are induced according to the invention. The data also show induction of CD8+ T cells, which are also very beneficial.
[0326] We refer to
[0327] For clarity please note that
[0328] It is a surprising benefit that the immunisations according to the present invention are also effective in raising/inducing antibody titers, despite the one-shot administration. It is a surprising benefit that the invention is as good as prior art compositions such as Zostavax for the induction of antibody responses.
Example 4: Prime-Boost Study
[0329] We demonstrate immunogenicity after prime-boost vaccination against VZV.
[0330] We refer to
[0331] 8 wk.sup.+ old female Balb/c mice were vaccinated intramuscularly with ChAdOX1-VZV-gE—group 1. 1.00E+07 iu ChAdOX1-VZV-gE then four weeks later boosted with 1.00E+07 iu ChAdOX1-VZV-gE ChAdOX2-VZV-gE—group 2. 1.00E+07 iu ChAdOX2-VZV-gE then four weeks later boosted with 1.00E+07 iu ChAdOX2-VZV-gE.
[0332] Zostavax—group 3. 1.29E+03 VZV Zostavax then four weeks later boosted with 1.29E+03 VZV Zostavax. Sera was taken at the indicated timepoints and assayed for anti-VZV-gpE specific antibodies.
[0333] Referring to
[0334] These data show that the single-shot or single-administration embodiments of the invention provide as good a response as a prime-boost regime. Thus it is an advantage of the invention that only a single shot or single dose (single administration) is needed.
Example 5: Comparative Study
[0335] The inventors compare the composition of the invention to prior art Shingrix™ across 3 doses and after one shot.
[0336] Humoral immunity after one-shot vaccination against VZV was tested. Groups of CD1 mice (n=7/8) were vaccinated intramuscularly with either ChAdOx1-VZVgpE or Shingrix™, at doses indicated (3 doses). Sera were collected at 4 weeks indicated after vaccination and the humoral immune response toward affinity purified glycoproteins of Varizella Zoster Virus (Strain Ellen) were measured by ELISA. We refer to
TABLE-US-00003 Vaccine Description Group Shingrix ™ low dose 1 0.2 μg/mouse Shingrix ™ mid dose 2 1 μg/mouse Shingrix ™ high dose 3 5 μg/mouse ChAdOx1-VZVgpE low 4 1*10{circumflex over ( )}6/mouse ChAdOx1-VZVgpE mid 5 1*10{circumflex over ( )}7/mouse ChAdOx1-VZVgpE high 6 1*10{circumflex over ( )}8/mouse
[0337] Cellular immunogenicity after one-shot vaccination against VZV was tested. Groups of CD1 mice (n=7/8) were vaccinated intramuscularly with either ChAdOx1-VZVgpE or Shingrix™, at doses indicated (3 doses). Splenocytes were collected 4 weeks after final vaccination and the cellular immune response against peptides spanning the whole glycoprotein-E were measured by ELISpot analysis. We refer to
TABLE-US-00004 Vaccine Description Group Shingrix ™ low dose 1 0.2 μg/mouse Shingrix ™ mid dose 2 1 μg/mouse Shingrix ™ high dose 3 5 μg/mouse ChAdOx1-VZVgpE low 4 1*10{circumflex over ( )}6/mouse ChAdOx1-VZVgpE mid 5 1*10{circumflex over ( )}7/mouse ChAdOx1-VZVgpE high 6 1*10{circumflex over ( )}8/mouse
Example 6: Comparative Study
[0338] Groups of outbred CD-1 mice (n=8) were vaccinated intramuscularly with [0339] Group 1; 1 ug of Shringrix with ASO1B adjuvant and four weeks later the animals were boosted with 1 ug of Shringrix with ASO1B adjuvant or [0340] Group 2; no prime and four weeks later the animals were vaccinated with 1.3×10.sup.3 pfu Zostavax or [0341] Group 3; 1.3×10.sup.3 pfu Zostavax and four weeks later animals were boosted with 1×10.sup.7 IU of ChAdOx1-VZVgpE or [0342] Group 4; no prime and four weeks later the animals were vaccinated 1×10.sup.7 IU of ChAdOx1-VZVgpE or [0343] Group 5; naïve animals.
[0344] We refer to
[0345] Serum was collected approximately three weeks after final vaccination and analysed for anti-VZVgpE antibodies.
[0346] Kruskal-Wallis analysis with Dunn's multiple comparison test demonstrates that Group 1; two shots of protein with adjuvant induces a significantly higher antibody titre when compared to Group 2; one shot of Zostavax or Group 4; one shot of ChAdOx1-VZVgpE, this result is as expected. However, there was no difference in the level of antibodies measured between Group 1 and Group 3. This is not expected, as ChAdOx1 after Zostavax would not be predicted to increase the humoral immune response to a comparable level of two shots of adjuvanted protein. This is an advantage, as currently UK adults aged 70 or over have been recommended to receive Zostavax vaccination, here we demonstrate that a boost vaccination of ChAdOx1-VZV-gpE can augment the antibody titres to those levels measured after two protein and adjuvant vaccinations, a regimen that is associated with efficacy of 91% or higher.
TABLE-US-00005 Dunn's multiple Adjusted P comparisons test Significant? Summary Value Shingrix 1 ug/mouse x2 vs. No ns >0.9999 Zostavax-ChAdOx1-gE Shingrix 1 ug/mouse x2 vs. Yes * 0.0172 ChAdOx1-gE x1 Shingrix 1 ug/mouse x2 vs. Yes ** 0.0019 Zostavax x1
Example 7: Comparative Study
[0347] Groups of C57BL6 mice (n=5) were vaccinated intramuscularly with
[0348] Group 1; 1 ug of Shringrix with ASO1B adjuvant and four weeks later the animals were boosted with 1 ug of Shringrix with ASO1B adjuvant or
[0349] Group 2; no prime and four weeks later the animals were vaccinated with 1.3×10.sup.3 pfu Zostavax or
[0350] Group 3; 1.3×10.sup.3 pfu Zostavax and four weeks later animals were boosted with 1×10.sup.7 IU of ChAdOx1-VZVgpE or
[0351] Group 4; no prime and four weeks later the animals were vaccinated 1×10.sup.7 IU of ChAdOx1-VZVgpE.
[0352] We refer to
[0353] Splenocytes were collected approximately four weeks after final vaccination and analysed for NK maturity and secretion of cytokines. Kruskal-Wallis analysis with Dunn's multiple comparison test demonstrates that NK cells after a prime-boost vaccination of 1.3×10.sup.3 pfu Zostax followed by 1×10.sup.7 IU of ChAdOx1-VZVgpE secrete more IFN-g (Group 3) when compared to the Shringix vaccination (Group 1). This is not expected and offers an advantage, NK cell activation has previously been demonstrated to augment the adaptive immune response and this strong induction of the innate immune response by ChAdOx1-VZVgpE after a Zostavax prime is not expected. Additionally, NK cells have been demonstrated to be critically important in mediating immunity against VZV infection (PMID; 2543925, 30565241).
TABLE-US-00006 Table of sequences SEQ ID NO: 1 wild type VZV GlyE amino acid sequence SEQ ID NO: 2 exemplary VZV GlyE amino acid sequence SEQ ID NO: 3 wild type nucleotide sequence encoding VZV GlyE (Genbank accession number AY253715.1; this is the nucleotide sequence encoding the amino acid sequence of accession number AAP32865.1 - SEQ ID NO: 1: (AAP32865.1 glycoprotein E [Human alphaherpesvirus 3])) SEQ ID NO: 4 exemplary nucleotide sequence encoding VZV GlyE codon-optimised for humans SEQ ID NO: 5 ChAdOx2: Viral vector based on Chimpanzee adenovirus C68 SEQ ID NO: 6 bgh polyadenylation signal SEQ ID NO: 7 exemplary sequence of long CMV promoter Please note that ‘Gly E’ means glycoprotein E, and is sometimes referred to as ‘gE’.
[0354] In one embodiment Gly E sequence having similarity to antigen insert SEQ ID NO: 1 of 50% or less may be used. In one embodiment Gly E sequence having similarity to antigen insert SEQ ID NO: 1 of 50% or more may be used, suitably 60% or more, suitably 70% or more, suitably 80% or more, suitably 90% or more, suitably 95% or more.
TABLE-US-00007 SEQUENCE LISTING SEQ ID NO: 1: exemplary Gly E sequence-GenBank accession number AAP32865.1- (>A7P32865.1 glycoprotein E [Human alphaherpesvirus 3])(also known as the amino acid sequence encoded by GenBank nucleotide accession number AY253715.1): MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSRKAYDHN SPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDRHKIVNVDQRQYGD VFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDV DCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYIWNMRGSDGTSTY ATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLAMHLQYKIHEAPFDLLLEWLYVPIDPTCQPM RLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDT PESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLA AVVLLCLVIFLICTAKRMRVKAYRVDKSPYNQSMYYAGLPVDDFEDSESTDTEEEFGNAIGGSHGGSSYTVYIDKTR SEQ ID NO: 2 exemplary VZV GlyE cassette amino acid sequence (sometimes referred to as “Insert-protein”): MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHA ESSWVNRGESSRKAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQM SAQEDLGDDTGIHVIPTLNGDDRHKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENH PFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAIQHICLKHTTCFQDVVVDVDCAENT KEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVLRTEKQYLGVYI WNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLA MHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHL AQRVASTVYQNCEHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVV YFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAA WTGGLAAVVLLCLVIFLICTAKRMRVKAYRVDKSPYNQSMYYAGLPVDDFEDSESTDTEE EFGNAIGGSHGGSSYTVYIDKTR SEQ ID NO: 3-wild type nucleotide sequence encoding VZV GlyE (Genbank accession number AY253715.1; this is the nucleotide sequence encoding the amino acid sequence of accession number AAP32865.1-SEQ ID NO: 1: (AAP32865.1 glycoprotein E [Human alphaherpesvirus 3])): ATGGGGACAGTTAATAAACCTGTGGTGGGGGTATTGATGGGGTTCGGAATTATCACGGGAACGTTGCGTATAACGAAT CCGGTCAGAGCATCCGTCTTGCGATACGATGATTTTCACATCGATGAAGACAAACTGGATACAAACTCCGTATATGAG CCTTACTACCATTCAGATCATGCGGAGTCTTCATGGGTAAATCGGGGAGAGTCTTCGCGAAAAGCGTACGATCATAAC TCACCTTATATATGGCCACGTAATGATTATGATGGATTTTTAGAGAACGCACACGAACACCATGGGGTGTATAATCAG GGCCGTGGTATCGATAGCGGGGAACGGTTAATGCAACCCACACAAATGTCTGCACAGGAGGATCTTGGGGACGATACG GGCATCCACGTTATCCCTACGTTAAACGGCGATGACAGACATAAAATTGTAAATGTGGACCAACGTCAATACGGTGAC GTGTTTAAAGGAGATCTTAATCCAAAACCCCAAGGCCAAAGACTCATTGAGGTGTCAGTGGAAGAAAATCACCCGTTT ACTTTACGCGCACCGATTCAGCGGATTTATGGAGTCCGGTACACCGAGACTTGGAGCTTTTTGCCGTCATTAACCTGT ACGGGAGACGCAGCGCCCGCCATCCAGCATATATGTTTAAAACATACAACATGCTTTCAAGACGTGGTGGTGGATGTG GATTGCGCGGAAAATACTAAAGAGGATCAGTTGGCCGAAATCAGTTACCGTTTTCAAGGTAAGAAGGAAGCGGACCAA CCGTGGATTGTTGTAAACACGAGCACACTGTTTGATGAACTCGAATTAGACCCCCCCGAGATTGAACCGGGTGTCTTG AAAGTACTTCGGACAGAAAAACAATACTTGGGTGTGTACATTTGGAACATGCGCGGCTCCGATGGTACGTCTACCTAC GCCACGTTTTTGGTCACCTGGAAAGGGGATGAAAAAACAAGAAACCCTACGCCCGCAGTAACTCCTCAACCAAGAGGG GCTGAGTTTCATATGTGGAATTACCACTCGCATGTATTTTCAGTTGGTGATACGTTTAGCTTGGCAATGCATCTTCAG TATAAGATACATGAAGCGCCATTTGATTTGCTGTTAGAGTGGTTGTATGTCCCCATCGATCCTACATGTCAACCAATG CGGTTATATTCTACGTGTTTGTATCATCCCAACGCACCCCAATGCCTCTCTCATATGAATTCCGGTTGTACATTTACC TCGCCACATTTAGCCCAGCGTGTTGCAAGCACAGTGTATCAAAATTGTGAACATGCAGATAACTACACCGCATATTGT CTGGGAATATCTCATATGGAGCCTAGCTTTGGTCTAATCTTACACGACGGGGGCACCACGTTAAAGTTTGTAGATACA CCCGAGAGTTTGTCGGGATTATACGTTTTTGTGGTGTATTTTAACGGGCATGTTGAAGCCGTAGCATACACTGTTGTA TCCACAGTAGATCATTTTGTAAACGCAATTGAAGAGCGTGGATTTCCGCCAACGGCCGGTCAGCCACCGGCGACTACT AAACCCAAGGAAATTACCCCCGTAAACCCCGGAACGTCACCACTTCTACGATATGCCGCATGGACCGGAGGGCTTGCA GCAGTAGTACTTTTATGTCTCGTAATATTTTTAATCTGTACGGCTAAACGAATGAGGGTTAAAGCCTATAGGGTAGAC AAGTCCCCGTATAACCAAAGCATGTATTACGCTGGCCTTCCAGTGGACGATTTCGAGGACTCGGAATCTACGGATACG GAAGAAGAGTTTGGTAACGCGATTGGAGGGAGTCACGGGGGTTCGAGTTACACGGTGTATATAGATAAGACCCGGTGA SEQ ID NO: 4-exemplary nucleotide sequence encoding VZV GlyE antigen cassette/expression cassette (sometimes referred to as “>Insert-Vaccine”) ATGGGCACCGTGAACAAGCCCGTCGTGGGCGTGCTGATGGGCTTCGGCATCATCACCGGCACCCTGCGGATCACCAAT CCTGTGCGGGCCAGCGTGCTGAGATACGACGACTTCCACATCGACGAGGACAAGCTGGACACCAACAGCGTGTACGAG CCCTACTACCACAGCGACCACGCCGAGAGCAGCTGGGTCAACAGAGGCGAGTCCAGCCGGAAGGCCTACGACCACAAC AGCCCCTACATCTGGCCCCGGAACGACTACGACGGCTTCCTGGAAAATGCCCACGAGCACCACGGCGTGTACAACCAG GGCAGAGGCATCGACAGCGGCGAGAGACTGATGCAGCCCACCCAGATGAGCGCCCAGGAAGATCTGGGCGACGACACC GGCATCCACGTGATCCCTACCCTGAACGGCGACGACCGGCACAAGATCGTGAACGTGGACCAGCGGCAGTACGGCGAC GTGTTCAAGGGCGACCTGAACCCCAAGCCCCAGGGACAGCGGCTGATTGAGGTGTCCGTGGAAGAGAACCACCCCTTC ACCCTGAGAGCCCCCATCCAGAGAATCTACGGCGTGCGCTATACCGAGACTTGGAGCTTCCTGCCCAGCCTGACCTGT ACTGGCGACGCCGCTCCTGCCATCCAGCACATCTGCCTGAAGCACACCACCTGTTTCCAGGACGTGGTGGTGGACGTG GACTGCGCCGAGAACACCAAAGAGGACCAGCTGGCCGAGATCAGCTACCGGTTCCAGGGCAAGAAAGAGGCCGACCAG CCCTGGATCGTCGTGAACACCAGCACCCTGTTCGACGAGCTGGAACTGGACCCCCCCGAGATTGAACCCGGGGTGCTG AAGGTGCTGCGGACCGAGAAGCAGTACCTGGGAGTGTACATCTGGAACATGCGGGGCAGCGACGGCACCTCTACCTAC GCCACCTTCCTCGTGACCTGGAAGGGCGACGAGAAAACCCGGAACCCTACCCCTGCCGTGACCCCTCAGCCTAGAGGC GCCGAGTTTCACATGTGGAATTACCACAGCCACGTGTTCAGCGTGGGCGACACCTTCTCCCTGGCCATGCATCTGCAG TACAAGATCCACGAGGCCCCCTTCGACCTGCTGCTGGAATGGCTGTACGTGCCCATCGACCCTACCTGCCAGCCCATG CGGCTGTACTCCACCTGTCTGTACCACCCCAACGCCCCCCAGTGCCTGAGCCACATGAATAGCGGCTGCACCTTCACC AGCCCCCACCTGGCTCAGAGGGTGGCCAGCACCGTGTACCAGAATTGCGAGCACGCCGACAACTACACCGCCTACTGC CTGGGCATCAGCCACATGGAACCTAGCTTCGGCCTGATCCTGCACGACGGCGGCACCACCCTGAAGTTCGTGGATACC CCAGAGAGCCTGAGCGGCCTGTACGTGTTCGTGGTGTACTTCAACGGCCACGTGGAAGCCGTGGCCTACACCGTGGTG TCCACCGTGGACCACTTCGTGAACGCCATCGAGGAACGGGGCTTCCCTCCAACTGCTGGACAGCCTCCTGCCACCACC AAGCCCAAAGAAATCACCCCCGTGAACCCCGGCACCAGCCCTCTGCTGCGCTATGCTGCTTGGACAGGCGGACTGGCT GCTGTGGTGCTGCTGTGCCTCGTGATTTTCCTGATCTGCACCGCCAAGCGGATGAGAGTGAAGGCCTATCGGGTGGAC AAGTCCCCCTACAACCAGAGCATGTACTACGCCGGCCTGCCCGTGGACGATTTCGAGGATAGCGAGAGCACCGACACC GAGGAAGAGTTCGGCAACGCCATTGGCGGCTCTCACGGCGGCAGCAGCTATACCGTGTACATCGACAAGACCCGCTGA SEQ ID NO: 5 ChAd0x2: Viral vector based on Chimpanzee adenovirus C68 ccatcttcaa taatatacct caaacttttt gtgcgcgtta atatgcaaat gaggcgtttg 60 aatttgggga ggaagggcgg tgattggtcg agggatgagc gaccgttagg ggcggggcga 120 gtgacgtttt gatgacgtgg ttgcgaggag gagccagttt gcaagttctc gtgggaaaag 180 tgacgtcaaa cgaggtgtgg tttgaacacg gaaatactca attttcccgc gctctctgac 240 aggaaatgag gtgtttctgg gcggatgcaa gtgaaaacgg gccattttcg cgcgaaaact 300 gaatgaggaa gtgaaaatct gagtaatttc gcgtttatgg cagggaggag tatttgccga 360 gggccgagta gactttgacc gattacgtgg gggtttcgat taccgtgttt ttcacctaaa 420 tttccgcgta cggtgtcaaa gtccggtgtt tttacgcgat cgctagcgac atcgatcaca 480 agtttgtaca aaaaagctga acgagaaacg taaaatgata taaatatcaa tatattaaat 540 tagattttgc ataaaaaaca gactacataa tactgtaaaa cacaacatat ccagtcacta 600 tggcggccgc cgatttattc aacaaagcca cgttgtgtct caaaatctct gatgttacat 660 tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa 720 tacaaggggt gttatgagcc atattcaacg ggaaacgtct tgctcgaggc cgcgattaaa 780 ttccaacatg gatgctgatt tatatgggta taaatgggct cgtgataatg tcgggcaatc 840 aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca 900 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac 960 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt 1020 actcaccact gcgatccccg ggaaaacagc attccaggta ttagaagaat atcctgattc 1080 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt 1140 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat 1200 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga 1260 acaagtctgg aaagaaatgc ataagctttt gccattctca ccggattcag tcgtcactca 1320 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga 1380 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct 1440 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc 1500 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt 1560 taattggttg taacactggc acgcgtggat ccggcttact aaaagccaga taacagtatg 1620 cgtatttgcg cgctgatttt tgcggtataa gaatatatac tgatatgtat acccgaagta 1680 tgtcaaaaag aggtatgcta tgaagcagcg tattacagtg acagttgaca gcgacagcta 1740 tcagttgctc aaggcatata tgatgtcaat atctccggtc tggtaagcac aaccatgcag 1800 aatgaagccc gtcgtctgcg tgccgaacgc tggaaagcgg aaaatcagga agggatggct 1860 gaggtcgccc ggtttattga aatgaacggc tcttttgctg acgagaacag gggctggtga 1920 aatgcagttt aaggtttaca cctataaaag agagagccgt tatcgtctgt ttgtggatgt 1980 acagagtgat attattgaca cgcccgggcg acggatggtg atccccctgg ccagtgcacg 2040 tctgctgtca gataaagtct cccgtgaact ttacccggtg gtgcatatcg gggatgaaag 2100 ctggcgcatg atgaccaccg atatggccag tgtgccggtc tccgttatcg gggaagaagt 2160 ggctgatctc agccaccgcg aaaatgacat caaaaacgcc attaacctga tgttctgggg 2220 aatataaatg tcaggctccc ttatacacag ccagtctgca ggtcgaccat agtgactgga 2280 tatgttgtgt tttacagtat tatgtagtct gttttttatg caaaatctaa tttaatatat 2340 tgatatttat atcattttac gtttctcgtt cagctttctt gtacaaagtg gtgatcgatt 2400 cgacagatcg cgatcgcaag tgagtagtgt tctggggcgg gggaggacct gcatgagggc 2460 cagaataact gaaatctgtg cttttctgtg tgttgcagca gcatgagcgg aagcggctcc 2520 tttgagggag gggtattcag cccttatctg acggggcgtc tcccctcctg ggcgggagtg 2580 cgtcagaatg tgatgggatc cacggtggac ggccggcccg tgcagcccgc gaactcttca 2640 accctgacct atgcaaccct gagctcttcg tcgttggacg cagctgccgc cgcagctgct 2700 gcatctgccg ccagcgccgt gcgcggaatg gccatgggcg ccggctacta cggcactctg 2760 gtggccaact cgagttccac caataatccc gccagcctga acgaggagaa gctgttgctg 2820 ctgatggccc agctcgaggc cttgacccag cgcctgggcg agctgaccca gcaggtggct 2880 cagctgcagg agcagacgcg ggccgcggtt gccacggtga aatccaaata aaaaatgaat 2940 caataaataa acggagacgg ttgttgattt taacacagag tctgaatctt tatttgattt 3000 ttcgcgcgcg gtaggccctg gaccaccggt ctcgatcatt gagcacccgg tggatctttt 3060 ccaggacccg gtagaggtgg gcttggatgt tgaggtacat gggcatgagc ccgtcccggg 3120 ggtggaggta gctccattgc agggcctcgt gctcgggggt ggtgttgtaa atcacccagt 3180 catagcaggg gcgcagggca tggtgttgca caatatcttt gaggaggaga ctgatggcca 3240 cgggcagccc tttggtgtag gtgtttacaa atctgttgag ctgggaggga tgcatgcggg 3300 gggagatgag gtgcatcttg gcctggatct tgagattggc gatgttaccg cccagatccc 3360 gcctggggtt catgttgtgc aggaccacca gcacggtgta tccggtgcac ttggggaatt 3420 tatcatgcaa cttggaaggg aaggcgtgaa agaatttggc gacgcctttg tgcccgccca 3480 ggttttccat gcactcatcc atgatgatgg cgatgggccc gtgggcggcg gcctgggcaa 3540 agacgtttcg ggggtcggac acatcatagt tgtggtcctg ggtgaggtca tcataggcca 3600 ttttaatgaa tttggggcgg agggtgccgg actgggggac aaaggtaccc tcgatcccgg 3660 gggcgtagtt cccctcacag atctgcatct cccaggcttt gagctcggag ggggggatca 3720 tgtccacctg cggggcgata aagaacacgg tttccggggc gggggagatg agctgggccg 3780 aaagcaagtt ccggagcagc tgggacttgc cgcagccggt ggggccgtag atgaccccga 3840 tgaccggctg caggtggtag ttgagggaga gacagctgcc gtcctcccgg aggagggggg 3900 ccacctcgtt catcatctcg cgcacgtgca tgttctcgcg caccagttcc gccaggaggc 3960 gctctccccc cagggatagg agctcctgga gcgaggcgaa gtttttcagc ggcttgagtc 4020 cgtcggccat gggcattttg gagagggttt gttgcaagag ttccaggcgg tcccagagct 4080 cggtgatgtg ctctacggca tctcgatcca gcagacctcc tcgtttcgcg ggttgggacg 4140 gctgcgggag tagggcacca gacgatgggc gtccagcgca gccagggtcc ggtccttcca 4200 gggtcgcagc gtccgcgtca gggtggtctc cgtcacggtg aaggggtgcg cgccgggctg 4260 ggcgcttgcg agggtgcgct tcaggctcat ccggctggtc gaaaaccgct cccgatcggc 4320 gccctgcgcg tcggccaggt agcaattgac catgagttcg tagttgagcg cctcggccgc 4380 gtggcctttg gcgcggagct tacctttgga agtctgcccg caggcgggac agaggaggga 4440 cttgagggcg tagagcttgg gggcgaggaa gacggactcg ggggcgtagg cgtccgcgcc 4500 gcagtgggcg cagacggtct cgcactccac gagccaggtg aggtcgggct ggtcggggtc 4560 aaaaaccagt ttcccgccgt tctttttgat gcgtttctta cctttggtct ccatgagctc 4620 gtgtccccgc tgggtgacaa agaggctgtc cgtgtccccg tagaccgact ttatgggccg 4680 gtcctcgagc ggtgtgccgc ggtcctcctc gtagaggaac cccgcccact ccgagacgaa 4740 agcccgggtc caggccagca cgaaggaggc cacgtgggac gggtagcggt cgttgtccac 4800 cagcgggtcc accttttcca gggtatgcaa acacatgtcc ccctcgtcca catccaggaa 4860 ggtgattggc ttgtaagtgt aggccacgtg accgggggtc ccggccgggg gggtataaaa 4920 gggtgcgggt ccctgctcgt cctcactgtc ttccggatcg ctgtccagga gcgccagctg 4980 ttggggtagg tattccctct cgaaggcggg catgacctcg gcactcaggt tgtcagtttc 5040 tagaaacgag gaggatttga tattgacggt gccggcggag atgcctttca agagcccctc 5100 gtccatctgg tcagaaaaga cgatcttttt gttgtcgagc ttggtggcga aggagccgta 5160 gagggcgttg gagaggagct tggcgatgga gcgcatggtc tggttttttt ccttgtcggc 5220 gcgctccttg gcggcgatgt tgagctgcac gtactcgcgc gccacgcact tccattcggg 5280 gaagacggtg gtcagctcgt cgggcacgat tctgacctgc cagccccgat tatgcagggt 5340 gatgaggtcc acactggtgg ccacctcgcc gcgcaggggc tcattagtcc agcagaggcg 5400 tccgcccttg cgcgagcaga aggggggcag ggggtccagc atgacctcgt cgggggggtc 5460 ggcatcgatg gtgaagatgc cgggcaggag gtcggggtca aagtagctga tggaagtggc 5520 cagatcgtcc agggcagctt gccattcgcg cacggccagc gcgcgctcgt agggactgag 5580 gggcgtgccc cagggcatgg gatgggtaag cgcggaggcg tacatgccgc agatgtcgta 5640 gacgtagagg ggctcctcga ggatgccgat gtaggtgggg tagcagcgcc ccccgcggat 5700 gctggcgcgc acgtagtcat acagctcgtg cgagggggcg aggagccccg ggcccaggtt 5760 ggtgcgactg ggcttttcgg cgcggtagac gatctggcgg aaaatggcat gcgagttgga 5820 ggagatggtg ggcctttgga agatgttgaa gtgggcgtgg ggcagtccga ccgagtcgcg 5880 gatgaagtgg gcgtaggagt cttgcagctt ggcgacgagc tcggcggtga ctaggacgtc 5940 cagagcgcag tagtcgaggg tctcctggat gatgtcatac ttgagctgtc ccttttgttt 6000 ccacagctcg cggttgagaa ggaactcttc gcggtccttc cagtactctt cgagggggaa 6060 cccgtcctga tctgcacggt aagagcctag catgtagaac tggttgacgg ccttgtaggc 6120 gcagcagccc ttctccacgg ggagggcgta ggcctgggcg gccttgcgca gggaggtgtg 6180 cgtgagggcg aaagtgtccc tgaccatgac cttgaggaac tggtgcttga agtcgatatc 6240 gtcgcagccc ccctgctccc agagctggaa gtccgtgcgc ttcttgtagg cggggttggg 6300 caaagcgaaa gtaacatcgt tgaagaggat cttgcccgcg cggggcataa agttgcgagt 6360 gatgcggaaa ggttggggca cctcggcccg gttgttgatg acctgggcgg cgagcacgat 6420 ctcgtcgaag ccgttgatgt tgtggcccac gatgtagagt tccacgaatc gcggacggcc 6480 cttgacgtgg ggcagtttct tgagctcctc gtaggtgagc tcgtcggggt cgctgagccc 6540 gtgctgctcg agcgcccagt cggcgagatg ggggttggcg cggaggaagg aagtccagag 6600 atccacggcc agggcggttt gcagacggtc ccggtactga cggaactgct gcccgacggc 6660 cattttttcg ggggtgacgc agtagaaggt gcgggggtcc ccgtgccagc gatcccattt 6720 gagctggagg gcgagatcga gggcgagctc gacgagccgg tcgtccccgg agagtttcat 6780 gaccagcatg aaggggacga gctgcttgcc gaaggacccc atccaggtgt aggtttccac 6840 atcgtaggtg aggaagagcc tttcggtgcg aggatgcgag ccgatgggga agaactggat 6900 ctcctgccac caattggagg aatggctgtt gatgtgatgg aagtagaaat gccgacggcg 6960 cgccgaacac tcgtgcttgt gtttatacaa gcggccacag tgctcgcaac gctgcacggg 7020 atgcacgtgc tgcacgagct gtacctgagt tcctttgacg aggaatttca gtgggaagtg 7080 gagtcgtggc gcctgcatct cgtgctgtac tacgtcgtgg tggtcggcct ggccctcttc 7140 tgcctcgatg gtggtcatgc tgacgagccc gcgcgggagg caggtccaga cctcggcgcg 7200 agcgggtcgg agagcgagga cgagggcgcg caggccggag ctgtccaggg tcctgagacg 7260 ctgcggagtc aggtcagtgg gcagcggcgg cgcgcggttg acttgcagga gtttttccag 7320 ggcgcgcggg aggtccagat ggtacttgat ctccaccgcg ccattggtgg cgacgtcgat 7380 ggcttgcagg gtcccgtgcc cctggggtgt gaccaccgtc ccccgtttct tcttgggcgg 7440 ctggggcgac gggggcggtg cctcttccat ggttagaagc ggcggcgagg acgcgcgccg 7500 ggcggcaggg gcggctcggg gcccggaggc aggggcggca ggggcacgtc ggcgccgcgc 7560 gcgggtaggt tctggtactg cgcccggaga agactggcgt gagcgacgac gcgacggttg 7620 acgtcctgga tctgacgcct ctgggtgaag gccacgggac ccgtgagttt gaacctgaaa 7680 gagagttcga cagaatcaat ctcggtatcg ttgacggcgg cctgccgcag gatctcttgc 7740 acgtcgcccg agttgtcctg gtaggcgatc tcggtcatga actgctcgat ctcctcctct 7800 tgaaggtctc cgcggccggc gcgctccacg gtggccgcga ggtcgttgga gatgcggccc 7860 atgagctgcg agaaggcgtt catgcccgcc tcgttccaga cgcggctgta gaccacgacg 7920 ccctcgggat cgccggcgcg catgaccacc tgggcgaggt tgagctccac gtggcgcgtg 7980 aagaccgcgt agttgcagag gcgctggtag aggtagttga gcgtggtggc gatgtgctcg 8040 gtgacgaaga aatacatgat ccagcggcgg agcggcatct cgctgacgtc gcccagcgcc 8100 tccaaacgtt ccatggcctc gtaaaagtcc acggcgaagt tgaaaaactg ggagttgcgc 8160 gccgagacgg tcaactcctc ctccagaaga cggatgagct cggcgatggt ggcgcgcacc 8220 tcgcgctcga aggcccccgg gagttcctcc acttcctctt cttcctcctc cactaacatc 8280 tcttctactt cctcctcagg cggcagtggt ggcgggggag ggggcctgcg tcgccggcgg 8340 cgcacgggca gacggtcgat gaagcgctcg atggtctcgc cgcgccggcg tcgcatggtc 8400 tcggtgacgg cgcgcccgtc ctcgcggggc cgcagcgtga agacgccgcc gcgcatctcc 8460 aggtggccgg gggggtcccc gttgggcagg gagagggcgc tgacgatgca tcttatcaat 8520 tgccccgtag ggactccgcg caaggacctg agcgtctcga gatccacggg atctgaaaac 8580 cgctgaacga aggcttcgag ccagtcgcag tcgcaaggta ggctgagcac ggtttcttct 8640 ggcgggtcat gttggttggg agcggggcgg gcgatgctgc tggtgatgaa gttgaaatag 8700 gcggttctga gacggcggat ggtggcgagg agcaccaggt ctttgggccc ggcttgctgg 8760 atgcgcagac ggtcggccat gccccaggcg tggtcctgac acctggccag gtccttgtag 8820 tagtcctgca tgagccgctc cacgggcacc tcctcctcgc ccgcgcggcc gtgcatgcgc 8880 gtgagcccga agccgcgctg gggctggacg agcgccaggt cggcgacgac gcgctcggcg 8940 aggatggctt gctggatctg ggtgagggtg gtctggaagt catcaaagtc gacgaagcgg 9000 tggtaggctc cggtgttgat ggtgtaggag cagttggcca tgacggacca gttgacggtc 9060 tggtggcccg gacgcacgag ctcgtggtac ttgaggcgcg agtaggcgcg cgtgtcgaag 9120 atgtagtcgt tgcaggtgcg caccaggtac tggtagccga tgaggaagtg cggcggcggc 9180 tggcggtaga gcggccatcg ctcggtggcg ggggcgccgg gcgcgaggtc ctcgagcatg 9240 gtgcggtggt agccgtagat gtacctggac atccaggtga tgccggcggc ggtggtggag 9300 gcgcgcggga actcgcggac gcggttccag atgttgcgca gcggcaggaa gtagttcatg 9360 gtgggcacgg tctggcccgt gaggcgcgcg cagtcgtgga tgctctatac gggcaaaaac 9420 gaaagcggtc agcggctcga ctccgtggcc tggaggctaa gcgaacgggt tgggctgcgc 9480 gtgtaccccg gttcgaatct cgaatcaggc tggagccgca gctaacgtgg tattggcact 9540 cccgtctcga cccaagcctg caccaaccct ccaggatacg gaggcgggtc gttttgcaac 9600 ttttttttgg aggccggatg agactagtaa gcgcggaaag cggccgaccg cgatggctcg 9660 ctgccgtagt ctggagaaga atcgccaggg ttgcgttgcg gtgtgccccg gttcgaggcc 9720 ggccggattc cgcggctaac gagggcgtgg ctgccccgtc gtttccaaga ccccatagcc 9780 agccgacttc tccagttacg gagcgagccc ctcttttgtt ttgtttgttt ttgccagatg 9840 catcccgtac tgcggcagat gcgcccccac caccctccac cgcaacaaca gccccctcca 9900 cagccggcgc ttctgccccc gccccagcag caacttccag ccacgaccgc cgcggccgcc 9960 gtgagcgggg ctggacagag ttatgatcac cagctggcct tggaagaggg cgaggggctg 10020 gcgcgcctgg gggcgtcgtc gccggagcgg cacccgcgcg tgcagatgaa aagggacgct 10080 cgcgaggcct acgtgcccaa gcagaacctg ttcagagaca ggagcggcga ggagcccgag 10140 gagatgcgcg cggcccggtt ccacgcgggg cgggagctgc ggcgcggcct ggaccgaaag 10200 agggtgctga gggacgagga tttcgaggcg gacgagctga cggggatcag ccccgcgcgc 10260 gcgcacgtgg ccgcggccaa cctggtcacg gcgtacgagc agaccgtgaa ggaggagagc 10320 aacttccaaa aatccttcaa caaccacgtg cgcaccctga tcgcgcgcga ggaggtgacc 10380 ctgggcctga tgcacctgtg ggacctgctg gaggccatcg tgcagaaccc caccagcaag 10440 ccgctgacgg cgcagctgtt cctggtggtg cagcatagtc gggacaacga agcgttcagg 10500 gaggcgctgc tgaatatcac cgagcccgag ggccgctggc tcctggacct ggtgaacatt 10560 ctgcagagca tcgtggtgca ggagcgcggg ctgccgctgt ccgagaagct ggcggccatc 10620 aacttctcgg tgctgagttt gggcaagtac tacgctagga agatctacaa gaccccgtac 10680 gtgcccatag acaaggaggt gaagatcgac gggttttaca tgcgcatgac cctgaaagtg 10740 ctgaccctga gcgacgatct gggggtgtac cgcaacgaca ggatgcaccg tgcggtgagc 10800 gccagcaggc ggcgcgagct gagcgaccag gagctgatgc atagtctgca gcgggccctg 10860 accggggccg ggaccgaggg ggagagctac tttgacatgg gcgcggacct gcactggcag 10920 cccagccgcc gggccttgga ggcggcggca ggaccctacg tagaagaggt ggacgatgag 10980 gtggacgagg agggcgagta cctggaagac tgatggcgcg accgtatttt tgctagatgc 11040 aacaacaaca gccacctcct gatcccgcga tgcgggcggc gctgcagagc cagccgtccg 11100 gcattaactc ctcggacgat tggacccagg ccatgcaacg catcatggcg ctgacgaccc 11160 gcaaccccga agcctttaga cagcagcccc aggccaaccg gctctcggcc atcctggagg 11220 ccgtggtgcc ctcgcgctcc aaccccacgc acgagaaggt cctggccatc gtgaacgcgc 11280 tggtggagaa caaggccatc cgcggcgacg aggccggcct ggtgtacaac gcgctgctgg 11340 agcgcgtggc ccgctacaac agcaccaacg tgcagaccaa cctggaccgc atggtgaccg 11400 acgtgcgcga ggccgtggcc cagcgcgagc ggttccaccg cgagtccaac ctgggatcca 11460 tggtggcgct gaacgccttc ctcagcaccc agcccgccaa cgtgccccgg ggccaggagg 11520 actacaccaa cttcatcagc gccctgcgcc tgatggtgac cgaggtgccc cagagcgagg 11580 tgtaccagtc cgggccggac tacttcttcc agaccagtcg ccagggcttg cagaccgtga 11640 acctgagcca ggctttcaag aacttgcagg gcctgtgggg cgtgcaggcc ccggtcgggg 11700 accgcgcgac ggtgtcgagc ctgctgacgc cgaactcgcg cctgctgctg ctgctggtgg 11760 cccccttcac ggacagcggc agcatcaacc gcaactcgta cctgggctac ctgattaacc 11820 tgtaccgcga ggccatcggc caggcgcacg tggacgagca gacctaccag gagatcaccc 11880 acgtgagccg cgccctgggc caggacgacc cgggcaacct ggaagccacc ctgaactttt 11940 tgctgaccaa ccggtcgcag aagatcccgc cccagtacgc gctcagcacc gaggaggagc 12000 gcatcctgcg ttacgtgcag cagagcgtgg gcctgttcct gatgcaggag ggggccaccc 12060 ccagcgccgc gctcgacatg accgcgcgca acatggagcc cagcatgtac gccagcaacc 12120 gcccgttcat caataaactg atggactact tgcatcgggc ggccgccatg aactctgact 12180 atttcaccaa cgccatcctg aatccccact ggctcccgcc gccggggttc tacacgggcg 12240 agtacgacat gcccgacccc aatgacgggt tcctgtggga cgatgtggac agcagcgtgt 12300 tctccccccg accgggtgct aacgagcgcc ccttgtggaa gaaggaaggc agcgaccgac 12360 gcccgtcctc ggcgctgtcc ggccgcgagg gtgctgccgc ggcggtgccc gaggccgcca 12420 gtcctttccc gagcttgccc ttctcgctga acagtatccg cagcagcgag ctgggcagga 12480 tcacgcgccc gcgcttgctg ggcgaagagg agtacttgaa tgactcgctg ttgagacccg 12540 agcgggagaa gaacttcccc aataacggga tagaaagcct ggtggacaag atgagccgct 12600 ggaagacgta tgcgcaggag cacagggacg atccccgggc gtcgcagggg gccacgagcc 12660 ggggcagcgc cgcccgtaaa cgccggtggc acgacaggca gcggggacag atgtgggacg 12720 atgaggactc cgccgacgac agcagcgtgt tggacttggg tgggagtggt aacccgttcg 12780 ctcacctgcg cccccgtatc gggcgcatga tgtaagagaa accgaaaata aatgatactc 12840 accaaggcca tggcgaccag cgtgcgttcg tttcttctct gttgttgttg tatctagtat 12900 gatgaggcgt gcgtacccgg agggtcctcc tccctcgtac gagagcgtga tgcagcaggc 12960 gatggcggcg gcggcgatgc agcccccgct ggaggctcct tacgtgcccc cgcggtacct 13020 ggcgcctacg gaggggcgga acagcattcg ttactcggag ctggcaccct tgtacgatac 13080 cacccggttg tacctggtgg acaacaagtc ggcggacatc gcctcgctga actaccagaa 13140 cgaccacagc aacttcctga ccaccgtggt gcagaacaat gacttcaccc ccacggaggc 13200 cagcacccag accatcaact ttgacgagcg ctcgcggtgg ggcggccagc tgaaaaccat 13260 catgcacacc aacatgccca acgtgaacga gttcatgtac agcaacaagt tcaaggcgcg 13320 ggtgatggtc tcccgcaaga cccccaatgg ggtgacagtg acagaggatt atgatggtag 13380 tcaggatgag ctgaagtatg aatgggtgga atttgagctg cccgaaggca acttctcggt 13440 gaccatgacc atcgacctga tgaacaacgc catcatcgac aattacttgg cggtggggcg 13500 gcagaacggg gtgctggaga gcgacatcgg cgtgaagttc gacactagga acttcaggct 13560 gggctgggac cccgtgaccg agctggtcat gcccggggtg tacaccaacg aggctttcca 13620 tcccgatatt gtcttgctgc ccggctgcgg ggtggacttc accgagagcc gcctcagcaa 13680 cctgctgggc attcgcaaga ggcagccctt ccaggaaggc ttccagatca tgtacgagga 13740 tctggagggg ggcaacatcc ccgcgctcct ggatgtcgac gcctatgaga aaagcaagga 13800 ggatgcagca gctgaagcaa ctgcagccgt agctaccgcc tctaccgagg tcaggggcga 13860 taattttgca agcgccgcag cagtggcagc ggccgaggcg gctgaaaccg aaagtaagat 13920 agtcattcag ccggtggaga aggatagcaa gaacaggagc tacaacgtac taccggacaa 13980 gataaacacc gcctaccgca gctggtacct agcctacaac tatggcgacc ccgagaaggg 14040 cgtgcgctcc tggacgctgc tcaccacctc ggacgtcacc tgcggcgtgg agcaagtcta 14100 ctggtcgctg cccgacatga tgcaagaccc ggtcaccttc cgctccacgc gtcaagttag 14160 caactacccg gtggtgggcg ccgagctcct gcccgtctac tccaagagct tcttcaacga 14220 gcaggccgtc tactcgcagc agctgcgcgc cttcacctcg cttacgcacg tcttcaaccg 14280 cttccccgag aaccagatcc tcgtccgccc gcccgcgccc accattacca ccgtcagtga 14340 aaacgttcct gctctcacag atcacgggac cctgccgctg cgcagcagta tccggggagt 14400 ccagcgcgtg accgttactg acgccagacg ccgcacctgc ccctacgtct acaaggccct 14460 gggcatagtc gcgccgcgcg tcctctcgag ccgcaccttc taaatgtcca ttctcatctc 14520 gcccagtaat aacaccggtt ggggcctgcg cgcgcccagc aagatgtacg gaggcgctcg 14580 ccaacgctcc acgcaacacc ccgtgcgcgt gcgcgggcac ttccgcgctc cctggggcgc 14640 cctcaagggc cgcgtgcggt cgcgcaccac cgtcgacgac gtgatcgacc aggtggtggc 14700 cgacgcgcgc aactacaccc ccgccgccgc gcccgtctcc accgtggacg ccgtcatcga 14760 cagcgtggtg gcggacgcgc gccggtacgc ccgcgccaag agccggcggc ggcgcatcgc 14820 ccggcggcac cggagcaccc ccgccatgcg cgcggcgcga gccttgctgc gcagggccag 14880 gcgcacggga cgcagggcca tgctcagggc ggccagacgc gcggcttcag gcgccagcgc 14940 cggcaggacc cggagacgcg cggccacggc ggcggcagcg gccatcgcca gcatgtcccg 15000 cccgcggcga gggaacgtgt actgggtgcg cgacgccgcc accggtgtgc gcgtgcccgt 15060 gcgcacccgc ccccctcgca cttgaagatg ttcacttcgc gatgttgatg tgtcccagcg 15120 gcgaggagga tgtccaagcg caaattcaag gaagagatgc tccaggtcat cgcgcctgag 15180 atctacggcc ctgcggtggt gaaggaggaa agaaagcccc gcaaaatcaa gcgggtcaaa 15240 aaggacaaaa aggaagaaga aagtgatgtg gacggattgg tggagtttgt gcgcgagttc 15300 gccccccggc ggcgcgtgca gtggcgcggg cggaaggtgc aaccggtgct gagacccggc 15360 accaccgtgg tcttcacgcc cggcgagcgc tccggcaccg cttccaagcg ctcctacgac 15420 gaggtgtacg gggatgatga tattctggag caggcggccg agcgcctggg cgagtttgct 15480 tacggcaagc gcagccgttc cgcaccgaag gaagaggcgg tgtccatccc gctggaccac 15540 ggcaacccca cgccgagcct caagcccgtg accttgcagc aggtgctgcc gaccgcggcg 15600 ccgcgccggg ggttcaagcg cgagggcgag gatctgtacc ccaccatgca gctgatggtg 15660 cccaagcgcc agaagctgga agacgtgctg gagaccatga aggtggaccc ggacgtgcag 15720 cccgaggtca aggtgcggcc catcaagcag gtggccccgg gcctgggcgt gcagaccgtg 15780 gacatcaaga ttcccacgga gcccatggaa acgcagaccg agcccatgat caagcccagc 15840 accagcacca tggaggtgca gacggatccc tggatgccat cggctcctag tcgaagaccc 15900 cggcgcaagt acggcgcggc cagcctgctg atgcccaact acgcgctgca tccttccatc 15960 atccccacgc cgggctaccg cggcacgcgc ttctaccgcg gtcataccag cagccgccgc 16020 cgcaagacca ccactcgccg ccgccgtcgc cgcaccgccg ctgcaaccac ccctgccgcc 16080 ctggtgcgga gagtgtaccg ccgcggccgc gcacctctga ccctgccgcg cgcgcgctac 16140 cacccgagca tcgccattta aactttcgcc agctttgcag atcaatggcc ctcacatgcc 16200 gccttcgcgt tcccattacg ggctaccgag gaagaaaacc gcgccgtaga aggctggcgg 16260 ggaacgggat gcgtcgccac caccaccggc ggcggcgcgc catcagcaag cggttggggg 16320 gaggcttcct gcccgcgctg atccccatca tcgccgcggc gatcggggcg atccccggca 16380 ttgcttccgt ggcggtgcag gcctctcagc gccactgaga cacacttgga aacatcttgt 16440 aataaaccca tggactctga cgctcctggt cctgtgatgt gttttcgtag acagatggaa 16500 gacatcaatt tttcgtccct ggctccgcga cacggcacgc ggccgttcat gggcacctgg 16560 agcgacatcg gcaccagcca actgaacggg ggcgccttca attggagcag tctctggagc 16620 gggcttaaga atttcgggtc cacgcttaaa acctatggca gcaaggcgtg gaacagcacc 16680 acagggcagg cgctgaggga taagctgaaa gagcagaact tccagcagaa ggtggtcgat 16740 gggctcgcct cgggcatcaa cggggtggtg gacctggcca accaggccgt gcagcggcag 16800 atcaacagcc gcctggaccc ggtgccgccc gccggctccg tggagatgcc gcaggtggag 16860 gaggagctgc ctcccctgga caagcggggc gagaagcgac cccgccccga tgcggaggag 16920 acgctgctga cgcacacgga cgagccgccc ccgtacgagg aggcggtgaa actgggtctg 16980 cccaccacgc ggcccatcgc gcccctggcc accggggtgc tgaaacccga aaagcccgcg 17040 accctggact tgcctcctcc ccagccttcc cgcccctcta cagtggctaa gcccctgccg 17100 ccggtggccg tggcccgcgc gcgacccggg ggcaccgccc gccctcatgc gaactggcag 17160 agcactctga acagcatcgt gggtctggga gtgcagagtg tgaagcgccg ccgctgctat 17220 taaacctacc gtagcgctta acttgcttgt ctgtgtgtgt atgtattatg tcgccgccgc 17280 cgctgtccac cagaaggagg agtgaagagg cgcgtcgccg agttgcaaga tggccacccc 17340 atcgatgctg ccccagtggg cgtacatgca catcgccgga caggacgctt cggagtacct 17400 gagtccgggt ctggtgcagt ttgcccgcgc cacagacacc tacttcagtc tggggaacaa 17460 gtttaggaac cccacggtgg cgcccacgca cgatgtgacc accgaccgca gccagcggct 17520 gacgctgcgc ttcgtgcccg tggaccgcga ggacaacacc tactcgtaca aagtgcgcta 17580 cacgctggcc gtgggcgaca accgcgtgct ggacatggcc agcacctact ttgacatccg 17640 cggcgtgctg gatcggggcc ctagcttcaa accctactcc ggcaccgcct acaacagtct 17700 ggcccccaag ggagcaccca acacttgtca gtggacatat aaagccgatg gtgaaactgc 17760 cacagaaaaa acctatacat atggaaatgc acccgtgcag ggcattaaca tcacaaaaga 17820 tggtattcaa cttggaactg acaccgatga tcagccaatc tacgcagata aaacctatca 17880 gcctgaacct caagtgggtg atgctgaatg gcatgacatc actggtactg atgaaaagta 17940 tggaggcaga gctcttaagc ctgataccaa aatgaagcct tgttatggtt cttttgccaa 18000 gcctactaat aaagaaggag gtcaggcaaa tgtgaaaaca ggaacaggca ctactaaaga 18060 atatgacata gacatggctt tctttgacaa cagaagtgcg gctgctgctg gcctagctcc 18120 agaaattgtt ttgtatactg aaaatgtgga tttggaaact ccagataccc atattgtata 18180 caaagcaggc acagatgaca gcagctcttc tattaatttg ggtcagcaag ccatgcccaa 18240 cagacctaac tacattggtt tcagagacaa ctttatcggg ctcatgtact acaacagcac 18300 tggcaatatg ggggtgctgg ccggtcaggc ttctcagctg aatgctgtgg ttgacttgca 18360 agacagaaac accgagctgt cctaccagct cttgcttgac tctctgggtg acagaacccg 18420 gtatttcagt atgtggaatc aggcggtgga cagctatgat cctgatgtgc gcattattga 18480 aaatcatggt gtggaggatg aacttcccaa ctattgtttc cctctggatg ctgttggcag 18540 aacagatact tatcagggaa ttaaggctaa tggaactgat caaaccacat ggaccaaaga 18600 tgacagtgtc aatgatgcta atgagatagg caagggtaat ccattcgcca tggaaatcaa 18660 catccaagcc aacctgtgga ggaacttcct ctacgccaac gtggccctgt acctgcccga 18720 ctcttacaag tacacgccgg ccaatgttac cctgcccacc aacaccaaca cctacgatta 18780 catgaacggc cgggtggtgg cgccctcgct ggtggactcc tacatcaaca tcggggcgcg 18840 ctggtcgctg gatcccatgg acaacgtgaa ccccttcaac caccaccgca atgcggggct 18900 gcgctaccgc tccatgctcc tgggcaacgg gcgctacgtg cccttccaca tccaggtgcc 18960 ccagaaattt ttcgccatca agagcctcct gctcctgccc gggtcctaca cctacgagtg 19020 gaacttccgc aaggacgtca acatgatcct gcagagctcc ctcggcaacg acctgcgcac 19080 ggacggggcc tccatctcct tcaccagcat caacctctac gccaccttct tccccatggc 19140 gcacaacacg gcctccacgc tcgaggccat gctgcgcaac gacaccaacg accagtcctt 19200 caacgactac ctctcggcgg ccaacatgct ctaccccatc ccggccaacg ccaccaacgt 19260 gcccatctcc atcccctcgc gcaactgggc cgccttccgc ggctggtcct tcacgcgtct 19320 caagaccaag gagacgccct cgctgggctc cgggttcgac ccctacttcg tctactcggg 19380 ctccatcccc tacctcgacg gcaccttcta cctcaaccac accttcaaga aggtctccat 19440 caccttcgac tcctccgtca gctggcccgg caacgaccgg ctcctgacgc ccaacgagtt 19500 cgaaatcaag cgcaccgtcg acggcgaggg ctacaacgtg gcccagtgca acatgaccaa 19560 ggactggttc ctggtccaga tgctggccca ctacaacatc ggctaccagg gcttctacgt 19620 gcccgagggc tacaaggacc gcatgtactc cttcttccgc aacttccagc ccatgagccg 19680 ccaggtggtg gacgaggtca actacaagga ctaccaggcc gtcaccctgg cctaccagca 19740 caacaactcg ggcttcgtcg gctacctcgc gcccaccatg cgccagggcc agccctaccc 19800 cgccaactac ccctacccgc tcatcggcaa gagcgccgtc accagcgtca cccagaaaaa 19860 gttcctctgc gacagggtca tgtggcgcat ccccttctcc agcaacttca tgtccatggg 19920 cgcgctcacc gacctcggcc agaacatgct ctatgccaac tccgcccacg cgctagacat 19980 gaatttcgaa gtcgacccca tggatgagtc cacccttctc tatgttgtct tcgaagtctt 20040 cgacgtcgtc cgagtgcacc agccccaccg cggcgtcatc gaggccgtct acctgcgcac 20100 ccccttctcg gccggtaacg ccaccaccta agctcttgct tcttgcaagc catggccgcg 20160 ggctccggcg agcaggagct cagggccatc atccgcgacc tgggctgcgg gccctacttc 20220 ctgggcacct tcgataagcg cttcccggga ttcatggccc cgcacaagct ggcctgcgcc 20280 atcgtcaaca cggccggccg cgagaccggg ggcgagcact ggctggcctt cgcctggaac 20340 ccgcgctcga acacctgcta cctcttcgac cccttcgggt tctcggacga gcgcctcaag 20400 cagatctacc agttcgagta cgagggcctg ctgcgccgca gcgccctggc caccgaggac 20460 cgctgcgtca ccctggaaaa gtccacccag accgtgcagg gtccgcgctc ggccgcctgc 20520 gggctcttct gctgcatgtt cctgcacgcc ttcgtgcact ggcccgaccg ccccatggac 20580 aagaacccca ccatgaactt gctgacgggg gtgcccaacg gcatgctcca gtcgccccag 20640 gtggaaccca ccctgcgccg caaccaggag gcgctctacc gcttcctcaa ctcccactcc 20700 gcctactttc gctcccaccg cgcgcgcatc gagaaggcca ccgccttcga ccgcatgaat 20760 caagacatgt aaaccgtgtg tgtatgttaa atgtctttaa taaacagcac tttcatgtta 20820 cacatgcatc tgagatgatt tatttagaaa tcgaaagggt tctgccgggt ctcggcatgg 20880 cccgcgggca gggacacgtt gcggaactgg tacttggcca gccacttgaa ctcggggatc 20940 agcagtttgg gcagcggggt gtcggggaag gagtcggtcc acagcttccg cgtcagttgc 21000 agggcgccca gcaggtcggg cgcggagatc ttgaaatcgc agttgggacc cgcgttctgc 21060 gcgcgggagt tgcggtacac ggggttgcag cactggaaca ccatcagggc cgggtgcttc 21120 acgctcgcca gcaccgtcgc gtcggtgatg ctctccacgt cgaggtcctc ggcgttggcc 21180 atcccgaagg gggtcatctt gcaggtctgc cttcccatgg tgggcacgca cccgggcttg 21240 tggttgcaat cgcagtgcag ggggatcagc atcatctggg cctggtcggc gttcatcccc 21300 gggtacatgg ccttcatgaa agcctccaat tgcctgaacg cctgctgggc cttggctccc 21360 tcggtgaaga agaccccgca ggacttgcta gagaactggt tggtggcgca cccggcgtcg 21420 tgcacgcagc agcgcgcgtc gttgttggcc agctgcacca cgctgcgccc ccagcggttc 21480 tgggtgatct tggcccggtc ggggttctcc ttcagcgcgc gctgcccgtt ctcgctcgcc 21540 acatccatct cgatcatgtg ctccttctgg atcatggtgg tcccgtgcag gcaccgcagc 21600 ttgccctcgg cctcggtgca cccgtgcagc cacagcgcgc acccggtgca ctcccagttc 21660 ttgtgggcga tctgggaatg cgcgtgcacg aagccctgca ggaagcggcc catcatggtg 21720 gtcagggtct tgttgctagt gaaggtcagc ggaatgccgc ggtgctcctc gttgatgtac 21780 aggtggcaga tgcggcggta cacctcgccc tgctcgggca tcagctggaa gttggctttc 21840 aggtcggtct ccacgcggta gcggtccatc agcatagtca tgatttccat acccttctcc 21900 caggccgaga cgatgggcag gctcataggg ttcttcacca tcatcttagc gctagcagcc 21960 gcggccaggg ggtcgctctc gtccagggtc tcaaagctcc gcttgccgtc cttctcggtg 22020 atccgcaccg gggggtagct gaagcccacg gccgccagct cctcctcggc ctgtctttcg 22080 tcctcgctgt cctggctgac gtcctgcagg accacatgct tggtcttgcg gggtttcttc 22140 ttgggcggca gcggcggcgg agatgttgga gatggcgagg gggagcgcga gttctcgctc 22200 accactacta tctcttcctc ttcttggtcc gaggccacgc ggcggtaggt atgtctcttc 22260 gggggcagag gcggaggcga cgggctctcg ccgccgcgac ttggcggatg gctggcagag 22320 ccccttccgc gttcgggggt gcgctcccgg cggcgctctg actgacttcc tccgcggccg 22380 gccattgtgt tctcctaggg aggaacaaca agcatggaga ctcagccatc gccaacctcg 22440 ccatctgccc ccaccgccga cgagaagcag cagcagcaga atgaaagctt aaccgccccg 22500 ccgcccagcc ccgccacctc cgacgcggcc gtcccagaca tgcaagagat ggaggaatcc 22560 atcgagattg acctgggcta tgtgacgccc gcggagcacg aggaggagct ggcagtgcgc 22620 ttttcacaag aagagataca ccaagaacag ccagagcagg aagcagagaa tgagcagagt 22680 caggctgggc tcgagcatga cggcgactac ctccacctga gcggggggga ggacgcgctc 22740 atcaagcatc tggcccggca ggccaccatc gtcaaggatg cgctgctcga ccgcaccgag 22800 gtgcccctca gcgtggagga gctcagccgc gcctacgagt tgaacctctt ctcgccgcgc 22860 gtgcccccca agcgccagcc caatggcacc tgcgagccca acccgcgcct caacttctac 22920 ccggtcttcg cggtgcccga ggccctggcc acctaccaca tctttttcaa gaaccaaaag 22980 atccccgtct cctgccgcgc caaccgcacc cgcgccgacg cccttttcaa cctgggtccc 23040 ggcgcccgcc tacctgatat cgcctccttg gaagaggttc ccaagatctt cgagggtctg 23100 ggcagcgacg agactcgggc cgcgaacgct ctgcaaggag aaggaggaga gcatgagcac 23160 cacagcgccc tggtcgagtt ggaaggcgac aacgcgcggc tggcggtgct caaacgcacg 23220 gtcgagctga cccatttcgc ctacccggct ctgaacctgc cccccaaagt catgagcgcg 23280 gtcatggacc aggtgctcat caagcgcgcg tcgcccatct ccgaggacga gggcatgcaa 23340 gactccgagg agggcaagcc cgtggtcagc gacgagcagc tggcccggtg gctgggtcct 23400 aatgctagtc cccagagttt ggaagagcgg cgcaaactca tgatggccgt ggtcctggtg 23460 accgtggagc tggagtgcct gcgccgcttc ttcgccgacg cggagaccct gcgcaaggtc 23520 gaggagaacc tgcactacct cttcaggcac gggttcgtgc gccaggcctg caagatctcc 23580 aacgtggagc tgaccaacct ggtctcctac atgggcatct tgcacgagaa ccgcctgggg 23640 cagaacgtgc tgcacaccac cctgcgcggg gaggcccggc gcgactacat ccgcgactgc 23700 gtctacctct acctctgcca cacctggcag acgggcatgg gcgtgtggca gcagtgtctg 23760 gaggagcaga acctgaaaga gctctgcaag ctcctgcaga agaacctcaa gggtctgtgg 23820 accgggttcg acgagcgcac caccgcctcg gacctggccg acctcatttt ccccgagcgc 23880 ctcaggctga cgctgcgcaa cggcctgccc gactttatga gccaaagcat gttgcaaaac 23940 tttcgctctt tcatcctcga acgctccgga atcctgcccg ccacctgctc cgcgctgccc 24000 tcggacttcg tgccgctgac cttccgcgag tgccccccgc cgctgtggag ccactgctac 24060 ctgctgcgcc tggccaacta cctggcctac cactcggacg tgatcgagga cgtcagcggc 24120 gagggcctgc tcgagtgcca ctgccgctgc aacctctgca cgccgcaccg ctccctggcc 24180 tgcaaccccc agctgctgag cgagacccag atcatcggca ccttcgagtt gcaagggccc 24240 agcgaaggcg agggttcagc cgccaagggg ggtctgaaac tcaccccggg gctgtggacc 24300 tcggcctact tgcgcaagtt cgtgcccgag gactaccatc ccttcgagat caggttctac 24360 gaggaccaat cccatccgcc caaggccgag ctgtcggcct gcgtcatcac ccagggggcg 24420 atcctggccc aattgcaagc catccagaaa tcccgccaag aattcttgct gaaaaagggc 24480 cgcggggtct acctcgaccc ccagaccggt gaggagctca accccggctt cccccaggat 24540 gccccgagga aacaagaagc tgaaagtgga gctgccgccc gtggaggatt tggaggaaga 24600 ctgggagaac agcagtcagg cagaggagga ggagatggag gaagactggg acagcactca 24660 ggcagaggag gacagcctgc aagacagtct ggaggaagac gaggaggagg cagaggagga 24720 ggtggaagaa gcagccgccg ccagaccgtc gtcctcggcg ggggagaaag caagcagcac 24780 ggataccatc tccgctccgg gtcggggtcc cgctcgacca cacagtagat gggacgagac 24840 cggacgattc ccgaacccca ccacccagac cggtaagaag gagcggcagg gatacaagtc 24900 ctggcggggg cacaaaaacg ccatcgtctc ctgcttgcag gcctgcgggg gcaacatctc 24960 cttcacccgg cgctacctgc tcttccaccg cggggtgaac tttccccgca acatcttgca 25020 ttactaccgt cacctccaca gcccctacta cttccaagaa gaggcagcag cagcagaaaa 25080 agaccagcag aaaaccagca gctagaaaat ccacagcggc ggcagcaggt ggactgagga 25140 tcgcggcgaa cgagccggcg caaacccggg agctgaggaa ccggatcttt cccaccctct 25200 atgccatctt ccagcagagt cgggggcagg agcaggaact gaaagtcaag aaccgttctc 25260 tgcgctcgct cacccgcagt tgtctgtatc acaagagcga agaccaactt cagcgcactc 25320 tcgaggacgc cgaggctctc ttcaacaagt actgcgcgct cactcttaaa gagtagcccg 25380 cgcccgccca gtcgcagaaa aaggcgggaa ttacgtcacc tgtgcccttc gccctagccg 25440 cctccaccca tcatcatgag caaagagatt cccacgcctt acatgtggag ctaccagccc 25500 cagatgggcc tggccgccgg tgccgcccag gactactcca cccgcatgaa ttggctcagc 25560 gccgggcccg cgatgatctc acgggtgaat gacatccgcg cccaccgaaa ccagatactc 25620 ctagaacagt cagcgctcac cgccacgccc cgcaatcacc tcaatccgcg taattggccc 25680 gccgccctgg tgtaccagga aattccccag cccacgaccg tactacttcc gcgagacgcc 25740 caggccgaag tccagctgac taactcaggt gtccagctgg cgggcggcgc caccctgtgt 25800 cgtcaccgcc ccgctcaggg tataaagcgg ctggtgatcc ggggcagagg cacacagctc 25860 aacgacgagg tggtgagctc ttcgctgggt ctgcgacctg acggagtctt ccaactcgcc 25920 ggatcgggga gatcttcctt cacgcctcgt caggccgtcc tgactttgga gagttcgtcc 25980 tcgcagcccc gctcgggtgg catcggcact ctccagttcg tggaggagtt cactccctcg 26040 gtctacttca accccttctc cggctccccc ggccactacc cggacgagtt catcccgaac 26100 ttcgacgcca tcagcgagtc ggtggacggc tacgattgag tttaaactca cccccttatc 26160 cagtgaaata aagatcatat tgatgatgat tttacagaaa taaaaaataa tcatttgatt 26220 tgaaataaag atacaatcat attgatgatt tgagtttaac aaaaaaataa agaatcactt 26280 acttgaaatc tgataccagg tctctgtcca tgttttctgc caacaccact tcactcccct 26340 cttcccagct ctggtactgc aggccccggc gggctgcaaa cttcctccac acgctgaagg 26400 ggatgtcaaa ttcctcctgt ccctcaatct tcattttatc ttctatcaga tgtccaaaaa 26460 gcgcgtccgg gtggatgatg acttcgaccc cgtctacccc tacgatgcag acaacgcacc 26520 gaccgtgccc ttcatcaacc cccccttcgt ctcttcagat ggattccaag agaagcccct 26580 gggggtgttg tccctgcgac tggccgaccc cgtcaccacc aagaacgggg aaatcaccct 26640 caagctggga gagggggtgg acctcgattc ctcgggaaaa ctcatctcca acacggccac 26700 caaggccgcc gcccctctca gtttttccaa caacaccatt tcccttaaca tggatcaccc 26760 cttttacact aaagatggaa aattatcctt acaagtttct ccaccattaa atatactgag 26820 aacaagcatt ctaaacacac tagctttagg ttttggatca ggtttaggac tccgtggctc 26880 tgccttggca gtacagttag tctctccact tacatttgat actgatggaa acataaagct 26940 taccttagac agaggtttgc atgttacaac aggagatgca attgaaagca acataagctg 27000 ggctaaaggt ttaaaatttg aagatggagc catagcaacc aacattggaa atgggttaga 27060 gtttggaagc agtagtacag aaacaggtgt tgatgatgct tacccaatcc aagttaaact 27120 tggatctggc cttagctttg acagtacagg agccataatg gctggtaaca aagaagacga 27180 taaactcact ttgtggacaa cacctgatcc atcaccaaac tgtcaaatac tcgcagaaaa 27240 tgatgcaaaa ctaacacttt gcttgactaa atgtggtagt caaatactgg ccactgtgtc 27300 agtcttagtt gtaggaagtg gaaacctaaa ccccattact ggcaccgtaa gcagtgctca 27360 ggtgtttcta cgttttgatg caaacggtgt tcttttaaca gaacattcta cactaaaaaa 27420 atactggggg tataggcagg gagatagcat agatggcact ccatatacca atgctgtagg 27480 attcatgccc aatttaaaag cttatccaaa gtcacaaagt tctactacta aaaataatat 27540 agtagggcaa gtatacatga atggagatgt ttcaaaacct atgcttctca ctataaccct 27600 caatggtact gatgacagca acagtacata ttcaatgtca ttttcataca cctggactaa 27660 tggaagctat gttggagcaa catttggggc taactcttat accttctcat acatcgccca 27720 agaatgaaca ctgtatccca ccctgcatgc caacccttcc caccccactc tgtggaacaa 27780 actctgaaac acaaaataaa ataaagttca agtgttttat tgattcaaca gtttcacaga 27840 accctagtat tcaacctgcc acctccctcc caacacacag agtacacagt cctttctccc 27900 cggctggcct taaaaagcat catatcatgg gtaacagaca tattcttagg tgttatattc 27960 cacacggttt cctgtcgagc caaacgctca tcagtgatat taataaactc cccgggcagc 28020 tcacttaagt tcatgtcgct gtccagctgc tgagccacag gctgctgtcc aacttgcggt 28080 tgcttaacgg gcggcgaagg agaagtccac gcctacatgg gggtagagtc ataatcgtgc 28140 atcaggatag ggcggtggtg ctgcagcagc gcgcgaataa actgctgccg ccgccgctcc 28200 gtcctgcagg aatacaacat ggcagtggtc tcctcagcga tgattcgcac cgcccgcagc 28260 ataaggcgcc ttgtcctccg ggcacagcag cgcaccctga tctcacttaa atcagcacag 28320 taactgcagc acagcaccac aatattgttc aaaatcccac agtgcaaggc gctgtatcca 28380 aagctcatgg cggggaccac agaacccacg tggccatcat accacaagcg caggtagatt 28440 aagtggcgac ccctcataaa cacgctggac ataaacatta cctcttttgg catgttgtaa 28500 ttcaccacct cccggtacca tataaacctc tgattaaaca tggcgccatc caccaccatc 28560 ctaaaccagc tggccaaaac ctgcccgccg gctatacact gcagggaacc gggactggaa 28620 caatgacagt ggagagccca ggactcgtaa ccatggatca tcatgctcgt catgatatca 28680 atgttggcac aacacaggca cacgtgcata cacttcctca ggattacaag ctcctcccgc 28740 gttagaacca tatcccaggg aacaacccat tcctgaatca gcgtaaatcc cacactgcag 28800 ggaagacctc gcacgtaact cacgttgtgc attgtcaaag tgttacattc gggcagcagc 28860 ggatgatcct ccagtatggt agcgcgggtt tctgtctcaa aaggaggtag acgatcccta 28920 ctgtacggag tgcgccgaga caaccgagat cgtgttggtc gtagtgtcat gccaaatgga 28980 acgccggacg tagtcatatt tcctgaagca aaaccaggtg cgggcgtgac aaacagatct 29040 gcgtctccgg tctcgccgct tagatcgctc tgtgtagtag ttgtagtata tccactctct 29100 caaagcatcc aggcgccccc tggcttcggg ttctatgtaa actccttcat gcgccgctgc 29160 cctgataaca tccaccaccg cagaataagc cacacccagc caacctacac attcgttctg 29220 cgagtcacac acgggaggag cgggaagagc tggaagaacc atgattaact ttattccaaa 29280 cggtctcgga gcacttcaaa atgcaggtcc cggaggtggc acctctcgcc cccactgtgt 29340 tggtggaaaa taacagccag gtcaaaggtg acacggttct cgagatgttc cacggtggct 29400 tccagcaaag cctccacgcg cacatccaga aacaagagga cagcgaaagc gggagcgttt 29460 tctaattcct caatcatcat attacactcc tgcaccatcc ccagataatt ttcatttttc 29520 cagccttgaa tgattcgtat tagttcctga ggtaaatcca agccagccat gataaaaagc 29580 tcgcgcagag cgccctccac cggcattctt aagcacaccc tcataattcc aagagattct 29640 gctcctggtt cacctgcagc agattaacaa tgggaatatc aaaatctctg ccgcgatccc 29700 taagctcctc cctcaacaat aactgtatgt aatctttcat atcatctccg aaatttttag 29760 ccatagggcc gccaggaata agagcagggc aagccacatt acagataaag cgaagtcctc 29820 cccagtgwgc attgccaaat gtaagattga aataagcatg ctggctagac cctgtgatat 29880 cttccagata actggacaga aaatcaggca agcaattttt aagaaaatca acaaaagaaa 29940 agtcgtccag gtgcaggttt agagcctcag gaacaacgat ggaataagtg caaggagtgc 30000 gttccagcat ggttagtgtt tttttggtga tctgtagaac aaaaaataaa catgcaatat 30060 taaaccatgc tagcctggcg aacaggtggg taaatcactc tttccagcac caggcaggct 30120 acggggtctc cggcgcgacc ctcgtagaag ctgtcgccat gattgaaaag catcaccgag 30180 agaccttccc ggtggccggc atggatgatt cgagaagaag catacactcc gggaacattg 30240 gcatccgtga gtgaaaaaaa gcgacctata aagcctcggg gcactacaat gctcaatctc 30300 aattccagca aagccacccc atgcggatgg agcacaaaat tggcaggtgc gtaaaaaatg 30360 taattactcc cctcctgcac aggcagcaaa gcccccgctc cctccagaaa cacatacaaa 30420 gcctcagcgt ccatagctta ccgagcacgg caggcgcaag agtcagagaa aaggctgagc 30480 tctaacctga ctgcccgctc ctgtgctcaa tatatagccc taacctacac tgacgtaaag 30540 gccaaagtct aaaaataccc gccaaataat cacacacgcc cagcacacgc ccagaaaccg 30600 gtgacacact caaaaaaata cgcgcacttc ctcaaacgcc caaaactgcc gtcatttccg 30660 ggttcccacg ctacgtcatc aaaacacgac tttcaaattc cgtcgaccgt taaaaacgtc 30720 acccgccccg cccctaacgg tcgcccgtct ctcagccaat cagcgccccg catccccaaa 30780 ttcaaacacc tcatttgcat attaacgcgc acaaaaagtt tgaggtatat tattgatgat 30840 gg 30842 SEQ ID NO: 6-bgh polyadenylation signal ctgtgccttctagttgccagccatctgttgtttgcccctcccccgtgccttccttgaccctggaaggtgcc actcccactgtcctttcctaataaaatgaggaaattgcatcgcattgtctgagtaggtgtcattctattct ggggggtggggtggggcaggacagcaagggggaggattgggaagacaatagcaggcatgctggggatgcgg tgggctctatgg SEQ ID NO: 7: long CMV promoter ATCGCCATTTTTCCAAAAGTGATTTTTGGGCATACGCGATATCTGGCGATAGCGCTTA TATCGTTTACGGGGGATGGCGATAGACGATTTGGTGACTTGGGCGATTCTGTGTGT CGCAAATATCGCATTTCGATATAGGTGACAGACGATATGAGGCTATATCGCCGATA GAGGCGACATCAAGCTGGCACATGGCCAATGCATATCGATCTATACATTGAATCAATA TTGGCCATTAGCCATATTATTCATTGGTTATATAGCATAAATCAATATTGGCTATTGG CCATTGCATACGTTGTATCCATATCATAATATGTACATTTATATTGGCTCATGTCCAAC ATTACCGCCATGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGG TCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCC CGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCC CATAGTAACGCCAATAGGGACTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAA ACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACG TCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTT TCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTT TGGCAGTACATCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTC CACCCCATTGACGTCAATGGGAGTTTGTTTGGCACCAAAATCAACGGGATTTCCAA AATGTCGTAACAACTCCGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTGGG AGGTCTATATAAGCAGAGCTCTCCCTATCAGTGATAGAGATCTCCCTATCAGTGATAG AGATCGTCGACGAGCTCGTTTAGTGAACCGTCAGATCGCCTGGAGACGCCATCCACG CTGTTTGACCTCCATAGAAGACACCGGGACCGATCCAGCCTCCGCGGCCGGGAACG GTGCATTGGAACGCGGATTCCCCGTGCCAAGAGTGACGTAAGTACCGCCTATAGAGT CTATAGGCCCACCCCCTTGGCTTCTTATGCATGCTATACTGTTTTTGGCTTGGGGTCT ATACACCCCCGCTTCCTCATGTTATAGGTGATGGTATAGCTTAGCCTATAGGTGTGGG TTATTGACCATTATTGACCACTCCCCTATTGGTGACGATACTTTCCATTACTAATCCAT AACATGGCTTTGCCACAACTCTTTATTGGCTATATGCCAATACACTGTCCTTCAG AGACTGACACGGACTCTGTATTTTTACAGGATGGGGTCTCATTATTATTTACAAATT CACATATACAACACCACCGTCCCCAGTGCCCGCAGTTTTTATTAAACATAACGTGGGA TCTCCACGCGAATCTCGGGTACGTGTTCCGGACATGGGCTCTTCTCCGGTAGCGGCG GAGCTTCTACATCCGAGCCCTGCTCCCATGCCTCCAGCGACTCATGGTCGCTCGGCAG CTCCTTGCTCCTAACAGTGGAGGCCAGACTTAGGCACAGCACGATGCCCACCACCACC AGTGTGCCGCACAAGGCCGTGGCGGTAGGGTATGTGTCTGAAAATGAGCTCGGGGA GCGGGCTTGCACCGCTGACGCATTTGGAAGACTTAAGGCAGCGGCAGAAGAAGATGC AGGCAGCTGAGTTGTTGTGTTCTGATAAGAGTCAGAGGTAACTCCCGTTGCGGTGCT GTTAACGGTGGAGGGCAGTGTAGTCTGAGCAGTACTCGTTGCTGCCGCGCGCGCCAC CAGACATAATAGCTGACAGACTAACAGACTGTTCCTTTCCATGGGTCTTTTCTGCA