质粒名称: | pLVX-EF1alpha-eGFP-2xStrep-IRES-Puro |
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出品公司: | Addgene |
目录编号: | 141395 |
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: | eGFP |
物种来源: | jellyfish |
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-eGFP-2xStrep-IRES-Puro was a gift from Nevan Krogan (Addgene plasmid # 141395 ; http://n2t.net/addgene:141395 ; RRID:Addgene_141395) |
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-eGFP-2xStrep-IRES-Puro 9642 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-eGFP-2xStrep-IRES-Puro SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9642) 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 9642) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..9642 /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 3544..4260 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 3544..4260 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3589..3610) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3850..3869) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4197..4218 /label=EGFP-C /note="EGFP, forward primer" CDS 4288..4311 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" CDS 4348..4371 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" misc_feature 4392..4965 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4559..4576) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 4786..4805 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 4998..5597 /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(4998..5017) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 5494..5514 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 5611..6199 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5664..5684) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(6082..6093) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 6406..7039 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" primer_bind complement(7167..7183) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7167..7183) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(7180..7202) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7191..7207 /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(7215..7245) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7260..7281 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7398..7415) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7569..8157) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7649..7668) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8328..9188) /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 8951..8970 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(9189..9293) /gene="bla" /label=AmpR promoter polyA_signal 9341..9475 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(9378..9397) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 9432..9451 /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 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg 3601 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc 3661 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc 3721 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg 3781 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc 3841 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc 3901 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg 3961 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag 4021 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc 4081 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac 4141 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg 4201 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag 4261 gtgcggccgc tcgagggagg cggtggatgg agccaccccc agttcgagaa gggcggcggc 4321 agcggcggcg gcagcggcgg cggcagctgg agccaccccc agttcgagaa ggggcccgtt 4381 taaggatccc gcccctctcc ctcccccccc cctaacgtta ctggccgaag ccgcttggaa 4441 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat 4501 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct 4561 ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct 4621 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc 4681 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa 4741 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc 4801 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg 4861 gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc 4921 ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataagcttg ccacaaccca 4981 caaggagacg accttccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg 5041 acgtcccccg ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc 5101 acaccgtcga cccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca 5161 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg 5221 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca 5281 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc 5341 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc 5401 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg 5461 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg 5521 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc 5581 gcaagcccgg tgcctagacg cgtctggaac aatcaacctc tggattacaa aatttgtgaa 5641 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta 5701 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa 5761 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg 5821 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc 5881 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc 5941 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg 6001 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg 6061 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg 6121 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc 6181 ctttgggccg cctccccgcc tggaattaat tctgcagtcg agacctagaa aaacatggag 6241 caatcacaag tagcaataca gcagctacca atgctgattg tgcctggcta gaagcacaag 6301 aggaggagga ggtgggtttt ccagtcacac ctcaggtacc tttaagacca atgacttaca 6361 aggcagctgt agatcttagc cactttttaa aagaaaagag gggactggaa gggctaattc 6421 actcccaacg aagacaagat atccttgatc tgtggatcta ccacacacaa ggctacttcc 6481 ctgattagca gaactacaca ccagggccag gggtcagata tccactgacc tttggatggt 6541 gctacaagct agtaccagtt gagccagata aggtagaaga ggccaataaa ggagagaaca 6601 ccagcttgtt acaccctgtg agcctgcatg ggatggatga cccggagaga gaagtgttag 6661 agtggaggtt tgacagccgc ctagcatttc atcacgtggc ccgagagctg catccggagt 6721 acttcaagaa ctgctgatat cgagcttgct acaagggact ttccgctggg gactttccag 6781 ggaggcgtgg cctgggcggg actggggagt ggcgagccct cagatcctgc atataagcag 6841 ctgctttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct 6901 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta 6961 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca 7021 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt 7081 gcaaagaaat gaatatcaga gagtgagagg ccttgacatt gctagcgttt accgtcgacc 7141 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg 7201 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa 7261 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac 7321 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt 7381 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga 7441 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca 7501 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg 7561 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt 7621 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc 7681 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct 7741 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc 7801 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta 7861 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca 7921 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag 7981 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag 8041 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt 8101 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa 8161 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg 8221 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga 8281 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta 8341 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc 8401 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg 8461 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga 8521 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt 8581 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt 8641 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc 8701 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc 8761 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca 8821 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag 8881 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg 8941 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa 9001 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa 9061 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga 9121 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga 9181 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg 9241 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt 9301 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc aacttgttta ttgcagctta 9361 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 9421 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcaactg 9481 gataactcaa gctaaccaaa atcatcccaa acttcccacc ccatacccta ttaccactgc 9541 caattacctg tggtttcatt tactctaaac ctgtgattcc tctgaattat tttcatttta 9601 aagaaattgt atttgttaaa tatgtactac aaacttagta gt //
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