质粒名称: | pLVX-EF1alpha-SARS-CoV-2-nsp7-2xStrep-IRES-Puro |
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出品公司: | Addgene |
目录编号: | 141373 |
存储实验室: | Nevan Krogan |
载体骨架基本信息 |
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载体名称: | pLVX-EF1alpha-IRES-Puro |
载体出品公司: | Takara Bio |
空载体大小: | 8825 bp |
完整质粒大小: | 9165 bp |
载体类型: | 哺乳细胞表达, Lentiviral |
筛选标记: | Puromycin |
载体抗性: | 氨苄霉素 |
生长温度: | 30 ℃ |
宿主菌: | NEB Stable |
拷贝数: | 未知 |
插入基因信息 |
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插入基因名称: | SARS-CoV-2-nsp7 |
物种来源: | SARS-CoV-2 |
基因突变: | human codon optimized |
启动子: | EF1alpha |
标签/融合蛋白: | 2xStrep (C terminal on insert) |
克隆方法: | 未知 |
文献引用方法 |
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材料和方法部分: |
pLVX-EF1alpha-SARS-CoV-2-nsp7-2xStrep-IRES-Puro was a gift from Nevan Krogan (Addgene plasmid # 141373 ; http://n2t.net/addgene:141373 ; RRID:Addgene_141373)
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参考文献部分: | A SARS-CoV-2 protein interaction map reveals targets for drug repurposing. Gordon DE, Jang GM, Bouhaddou M, Xu J, Obernier K, White KM, O'Meara MJ, Rezelj VV, Guo JZ, Swaney DL, Tummino TA, Huttenhain R, Kaake RM, Richards AL, Tutuncuoglu B, Foussard H, Batra J, Haas K, Modak M, Kim M, Haas P, Polacco BJ, Braberg H, Fabius JM, Eckhardt M, Soucheray M, Bennett MJ, Cakir M, McGregor MJ, Li Q, Meyer B, Roesch F, Vallet T, Mac Kain A, Miorin L, Moreno E, Naing ZZC, Zhou Y, Peng S, Shi Y, Zhang Z, Shen W, Kirby IT, Melnyk JE, Chorba JS, Lou K, Dai SA, Barrio-Hernandez I, Memon D, Hernandez-Armenta C, Lyu J, Mathy CJP, Perica T, Pilla KB, Ganesan SJ, Saltzberg DJ, Rakesh R, Liu X, Rosenthal SB, Calviello L, Venkataramanan S, Liboy-Lugo J, Lin Y, Huang XP, Liu Y, Wankowicz SA, Bohn M, Safari M, Ugur FS, Koh C, Savar NS, Tran QD, Shengjuler D, Fletcher SJ, O'Neal MC, Cai Y, Chang JCJ, Broadhurst DJ, Klippsten S, Sharp PP, Wenzell NA, Kuzuoglu-Ozturk D, Wang HY, Trenker R, Young JM, Cavero DA, Hiatt J, Roth TL, Rathore U, Subramanian A, Noack J, Hubert M, Stroud RM, Frankel AD, Rosenberg OS, Verba KA, Agard DA, Ott M, Emerman M, Jura N, von Zastrow M, Verdin E, Ashworth A, Schwartz O, d'Enfert C, Mukherjee S, Jacobson M, Malik HS, Fujimori DG, Ideker T, Craik CS, Floor SN, Fraser JS, Gross JD, Sali A, Roth BL, Ruggero D, Taunton J, Kortemme T, Beltrao P, Vignuzzi M, Garcia-Sastre A, Shokat KM, Shoichet BK, Krogan NJ. Nature. 2020 Jul;583(7816):459-468. doi: 10.1038/s41586-020-2286-9. Epub 2020 Apr 30. 10.1038/s41586-020-2286-9 PubMed 32353859 |
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载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pLVX-EF1alpha-SARS-CoV-2-nsp7-2xStrep-IRES-Puro 9165 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Lentiviral expression of SARS-CoV-2 protein; transient expression and generate lentivirus. ACCESSION . VERSION . KEYWORDS pLVX-EF1alpha-SARS-CoV-2-nsp7-2xStrep-IRES-Puro SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9165) AUTHORS Gordon DE, et al. TITLE A SARS-CoV-2-Human Protein-Protein Interaction Map Reveals Drug Targets and Potential Drug Repurposing JOURNAL BioRxiv March 2020 REFERENCE 2 (bases 1 to 9165) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..9165 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 1..634 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 681..806 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1303..1536 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1721..1765 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 2028..2143 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" promoter 2338..3519 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 2568..3510 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 3467..3487 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" CDS 3820..3843 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" CDS 3880..3903 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" misc_feature 3915..4488 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4082..4099) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 4309..4328 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 4521..5120 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(4521..4540) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 5017..5037 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 5134..5722 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5187..5207) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5605..5616) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 5929..6562 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" primer_bind complement(6690..6706) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6690..6706) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6703..6725) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6714..6730 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(6738..6768) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6783..6804 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6921..6938) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7092..7680) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7172..7191) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7851..8711) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 8474..8493 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8712..8816) /gene="bla" /label=AmpR promoter polyA_signal 8864..8998 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(8901..8920) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 8955..8974 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag 2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac 2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg 2161 acagcagaga tccagtttat cgatgagtaa ttcatacaaa aggactcgcc cctgccttgg 2221 ggaatcccag ggaccgtcgt taaactccca ctaacgtaga acccagagat cgctgcgttc 2281 ccgccccctc acccgcccgc tctcgtcatc actgaggtgg agaagagcat gcgtgaggct 2341 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag 2401 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg 2461 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag 2521 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt 2581 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt 2641 ccacgcccct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg 2701 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg 2761 cttgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct 2821 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg 2881 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc 2941 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga 3001 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct 3061 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca 3121 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg 3181 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg 3241 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat 3301 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg 3361 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa 3421 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca 3481 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gatctatttc cggtgaattc 3541 gccgccacca tgagcaagat gagcgacgta aaatgcacgt cagttgttct cttgagcgtc 3601 ctgcaacaac tcagagtgga gtctagttcc aagctctggg ctcaatgcgt tcagcttcac 3661 aatgacattc tgttggcgaa agataccacg gaggcattcg agaagatggt ctcacttctg 3721 agcgttctcc tgtcaatgca aggggcggtc gatatcaata aattgtgtga ggaaatgctc 3781 gacaaccggg cgacattgca gctcgaaggc ggcgggggat ggagccatcc acaattcgag 3841 aaaggcggtg gttcaggagg aggtagcggg ggtggatcat ggtcacatcc gcagtttgaa 3901 aagtaaggat cccgcccctc tccctccccc ccccctaacg ttactggccg aagccgcttg 3961 gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc 4021 aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc 4081 cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa 4141 gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct 4201 ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca 4261 caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca 4321 agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat 4381 ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc 4441 cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataagc ttgccacaac 4501 ccacaaggag acgaccttcc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg 4561 acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc 4621 gccacaccgt cgacccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc 4681 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg 4741 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc 4801 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg 4861 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc 4921 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg 4981 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc 5041 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga 5101 cccgcaagcc cggtgcctag acgcgtctgg aacaatcaac ctctggatta caaaatttgt 5161 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct 5221 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat 5281 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg 5341 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag 5401 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc 5461 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg 5521 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc 5581 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc 5641 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc 5701 tccctttggg ccgcctcccc gcctggaatt aattctgcag tcgagaccta gaaaaacatg 5761 gagcaatcac aagtagcaat acagcagcta ccaatgctga ttgtgcctgg ctagaagcac 5821 aagaggagga ggaggtgggt tttccagtca cacctcaggt acctttaaga ccaatgactt 5881 acaaggcagc tgtagatctt agccactttt taaaagaaaa gaggggactg gaagggctaa 5941 ttcactccca acgaagacaa gatatccttg atctgtggat ctaccacaca caaggctact 6001 tccctgatta gcagaactac acaccagggc caggggtcag atatccactg acctttggat 6061 ggtgctacaa gctagtacca gttgagccag ataaggtaga agaggccaat aaaggagaga 6121 acaccagctt gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtgt 6181 tagagtggag gtttgacagc cgcctagcat ttcatcacgt ggcccgagag ctgcatccgg 6241 agtacttcaa gaactgctga tatcgagctt gctacaaggg actttccgct ggggactttc 6301 cagggaggcg tggcctgggc gggactgggg agtggcgagc cctcagatcc tgcatataag 6361 cagctgcttt ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc 6421 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa 6481 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag 6541 tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa 6601 cttgcaaaga aatgaatatc agagagtgag aggccttgac attgctagcg tttaccgtcg 6661 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat 6721 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc 6781 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga 6841 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 6901 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 6961 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 7021 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 7081 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 7141 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 7201 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 7261 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag 7321 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 7381 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 7441 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg 7501 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg 7561 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct 7621 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa 7681 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 7741 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa 7801 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc 7861 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga 7921 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca 7981 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc 8041 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat 8101 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc 8161 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt 8221 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc 8281 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg 8341 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt 8401 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg 8461 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga 8521 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg 8581 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg 8641 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt 8701 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc 8761 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca 8821 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atcaacttgt ttattgcagc 8881 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 8941 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcaa 9001 ctggataact caagctaacc aaaatcatcc caaacttccc accccatacc ctattaccac 9061 tgccaattac ctgtggtttc atttactcta aacctgtgat tcctctgaat tattttcatt 9121 ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagt //
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