质粒名称: | pLVX-EF1alpha-SARS-CoV-2-orf3a-2xStrep-IRES-Puro |
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出品公司: | Addgene |
目录编号: | 141383 |
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: | 8825 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-orf3a |
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-orf3a-2xStrep-IRES-Puro was a gift from Nevan Krogan (Addgene plasmid # 141383 ; http://n2t.net/addgene:141383 ; RRID:Addgene_141383)
For viral preps, please replace (Addgene plasmid # 141383) in the above sentence with: (Addgene viral prep # 141383-LV)
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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-orf3a-2xStrep-IRES-Puro 9738 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-orf3a-2xStrep-IRES-Puro SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9738) 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 9738) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..9738 /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" regulatory 3544..3553 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 4393..4416 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" CDS 4453..4476 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" misc_feature 4488..5061 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4655..4672) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 4882..4901 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5094..5693 /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(5094..5113) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 5590..5610 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 5707..6295 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5760..5780) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(6178..6189) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 6502..7135 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" primer_bind complement(7263..7279) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7263..7279) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(7276..7298) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7287..7303 /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(7311..7341) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7356..7377 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7494..7511) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7665..8253) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7745..7764) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8424..9284) /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 9047..9066 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(9285..9389) /gene="bla" /label=AmpR promoter polyA_signal 9437..9571 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(9474..9493) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 9528..9547 /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 tggacctttt tatgagaatc tttaccatcg ggactgtcac actgaaacag 3601 ggagaaatta aggacgcgac cccgagtgac tttgtgaggg ccactgctac aattcccatc 3661 caagcctctc ttccattcgg ctggctcata gtgggtgtgg cactgctcgc ggtattccag 3721 tcagcgtcta agatcataac gctgaagaag agatggcagt tggctctcag caaaggtgtt 3781 catttcgtct gtaacctgct tcttcttttc gtcactgtct attctcactt gctcttggtt 3841 gcagccgggc tggaagcccc gtttctgtat ctgtacgcgc tcgtttactt cctccagagc 3901 ataaatttcg tacgaattat tatgagactc tggttgtgct ggaagtgtcg gagcaagaat 3961 cctctcttgt atgacgccaa ttacttcctt tgttggcaca ctaattgtta tgattactgc 4021 ataccttaca atagcgttac ctcatccata gtaatcacct ccggtgacgg aacaacgtca 4081 ccaatctccg agcatgatta ccagattgga ggctatacgg aaaaatggga gagtggtgtc 4141 aaggactgtg tggtgctgca tagctatttc acaagcgatt attatcaact gtacagtaca 4201 cagttgtcca cggataccgg ggtcgagcat gtcacatttt ttatttataa caagatcgtt 4261 gatgagccag aggagcatgt gcagatccac acgatagatg gaagctccgg tgtagtgaat 4321 ccagtcatgg aaccgatcta tgacgagccc accacgacca catctgtgcc cctgctcgaa 4381 ggcggcgggg gatggagcca tccacaattc gagaaaggcg gtggttcagg aggaggtagc 4441 gggggtggat catggtcaca tccgcagttt gaaaagtaag gatcccgccc ctctccctcc 4501 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat 4561 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct 4621 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg 4681 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta 4741 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag 4801 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg 4861 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat 4921 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca 4981 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc 5041 ctttgaaaaa cacgatgata agcttgccac aacccacaag gagacgacct tccatgaccg 5101 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc 5161 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca 5221 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca 5281 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg 5341 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc 5401 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt 5461 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg 5521 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct 5581 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg 5641 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tagacgcgtc 5701 tggaacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat 5761 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct 5821 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag 5881 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc 5941 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc 6001 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct 6061 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg 6121 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg 6181 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg 6241 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga 6301 attaattctg cagtcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag 6361 ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag 6421 tcacacctca ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact 6481 ttttaaaaga aaagagggga ctggaagggc taattcactc ccaacgaaga caagatatcc 6541 ttgatctgtg gatctaccac acacaaggct acttccctga ttagcagaac tacacaccag 6601 ggccaggggt cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc 6661 cagataaggt agaagaggcc aataaaggag agaacaccag cttgttacac cctgtgagcc 6721 tgcatgggat ggatgacccg gagagagaag tgttagagtg gaggtttgac agccgcctag 6781 catttcatca cgtggcccga gagctgcatc cggagtactt caagaactgc tgatatcgag 6841 cttgctacaa gggactttcc gctggggact ttccagggag gcgtggcctg ggcgggactg 6901 gggagtggcg agccctcaga tcctgcatat aagcagctgc tttttgcctg tactgggtct 6961 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 7021 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 7081 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt 7141 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt 7201 gagaggcctt gacattgcta gcgtttaccg tcgacctcta gctagagctt ggcgtaatca 7261 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga 7321 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt 7381 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga 7441 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc 7501 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg 7561 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc 7621 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 7681 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 7741 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 7801 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat 7861 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 7921 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 7981 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 8041 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 8101 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 8161 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag 8221 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg 8281 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa 8341 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata 8401 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg 8461 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata 8521 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg 8581 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct 8641 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt 8701 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc 8761 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga 8821 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt 8881 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc 8941 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa 9001 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca 9061 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca 9121 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct 9181 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc 9241 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa 9301 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt 9361 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc 9421 gacggatcgg gagatcaact tgtttattgc agcttataat ggttacaaat aaagcaatag 9481 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 9541 actcatcaat gtatcttatc atgtctggat caactggata actcaagcta accaaaatca 9601 tcccaaactt cccaccccat accctattac cactgccaat tacctgtggt ttcatttact 9661 ctaaacctgt gattcctctg aattattttc attttaaaga aattgtattt gttaaatatg 9721 tactacaaac ttagtagt //
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