质粒名称: | pLVX-EF1alpha-SARS-CoV-2-orf9c-IRES-Puro |
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出品公司: | Addgene |
目录编号: | 141393 |
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: | 8825bp |
完整质粒大小: | 9126 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-orf9c |
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 (N 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: | pLXV-EF1alpha-2xStrep-SARS-CoV-2-orf9c-IRES-Puro was a gift from Nevan Krogan (Addgene plasmid # 141393 ; http://n2t.net/addgene:141393 ; RRID:Addgene_141393) |
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 pLXV-EF1alpha-2xStrep-SARS-CoV-2-orf9c-IRES-Puro 9126 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Lentiviral expression of SARS-CoV-2 protein; transient expression and generate lentivirus. ACCESSION . VERSION . KEYWORDS pLXV-EF1alpha-2xStrep-SARS-CoV-2-orf9c-IRES-Puro SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9126) 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 9126) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..9126 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..1182 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 231..1173 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1130..1150 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" CDS 1216..1239 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" CDS 1276..1299 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" misc_feature 1539..2112 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(1706..1723) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 1933..1952 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 2145..2744 /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(2145..2164) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 2641..2661 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 2758..3346 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2811..2831) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3229..3240) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 3553..4186 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" primer_bind complement(4314..4330) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4314..4330) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4327..4349) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4338..4354 /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(4362..4392) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4407..4428 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4545..4562) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4716..5304) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4796..4815) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5475..6335) /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 6098..6117 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6336..6440) /gene="bla" /label=AmpR promoter polyA_signal 6488..6622 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6525..6544) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6579..6598 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" LTR 6790..7423 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 7470..7595 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 8092..8325 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 8510..8554 /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 8817..8932 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" ORIGIN 1 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg 61 gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg 121 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag 181 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg 241 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta 301 cttccacgcc cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg 361 tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc 421 tggcttgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct 481 gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc 541 tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg 601 gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg 661 cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg 721 cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca 781 ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg 841 aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt 901 ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc 961 gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg 1021 atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg gcacttgatg 1081 taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag 1141 acagtggttc aaagtttttt tcttccattt caggtgtcgt gaggatctat ttccggtgaa 1201 ttcgccgcca ccatgtggtc ccatccgcag tttgagaagg gtggtggttc aggcggaggc 1261 tccgggggcg ggagctggtc tcacccgcaa tttgaaaaag gcgctgcggc tcttcaatcc 1321 tgctataact tcttgaaaga gcaacactgc caaaaggcga gcacacagaa aggagccgag 1381 gcagccgtaa agccgcttct tgtccctcat cacgttgtcg ccacagtcca agaaatacag 1441 cttcaagcgg ccgttggcga gcttctgctc ctcgaatggc tcgcaatggc ggtgatgttg 1501 ttgctgttgt gctgctgtct tacggattga ggatcccgcc cctctccctc ccccccccct 1561 aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt 1621 tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg 1681 acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc 1741 gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt 1801 tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta 1861 taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg 1921 gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag 1981 gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag 2041 tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa 2101 acacgatgat aagcttgcca caacccacaa ggagacgacc ttccatgacc gagtacaagc 2161 ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg 2221 cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg 2281 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg 2341 tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg 2401 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc 2461 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg 2521 ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc 2581 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc 2641 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg 2701 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctagacgcgt ctggaacaat 2761 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct 2821 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg 2881 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg 2941 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt 3001 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt 3061 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg 3121 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc 3181 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat 3241 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc 3301 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg aattaattct 3361 gcagtcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca gctaccaatg 3421 ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca gtcacacctc 3481 aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag 3541 aaaagagggg actggaaggg ctaattcact cccaacgaag acaagatatc cttgatctgt 3601 ggatctacca cacacaaggc tacttccctg attagcagaa ctacacacca gggccagggg 3661 tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag ccagataagg 3721 tagaagaggc caataaagga gagaacacca gcttgttaca ccctgtgagc ctgcatggga 3781 tggatgaccc ggagagagaa gtgttagagt ggaggtttga cagccgccta gcatttcatc 3841 acgtggcccg agagctgcat ccggagtact tcaagaactg ctgatatcga gcttgctaca 3901 agggactttc cgctggggac tttccaggga ggcgtggcct gggcgggact ggggagtggc 3961 gagccctcag atcctgcata taagcagctg ctttttgcct gtactgggtc tctctggtta 4021 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa 4081 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac 4141 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt 4201 catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggcct 4261 tgacattgct agcgtttacc gtcgacctct agctagagct tggcgtaatc atggtcatag 4321 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc 4381 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc 4441 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa 4501 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg 4561 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 4621 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 4681 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 4741 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 4801 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 4861 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc 4921 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 4981 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 5041 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 5101 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca 5161 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 5221 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 5281 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 5341 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 5401 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 5461 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 5521 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 5581 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 5641 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 5701 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 5761 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt 5821 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 5881 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 5941 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 6001 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 6061 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga 6121 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 6181 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 6241 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 6301 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 6361 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 6421 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg 6481 ggagatcaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa 6541 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa 6601 tgtatcttat catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact 6661 tcccacccca taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg 6721 tgattcctct gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa 6781 cttagtagtt ggaagggcta attcactccc aaagaagaca agatatcctt gatctgtgga 6841 tctaccacac acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca 6901 gatatccact gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag 6961 aagaggccaa taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg 7021 atgacccgga gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg 7081 tggcccgaga gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg 7141 gactttccgc tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag 7201 ccctcagatc ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac 7261 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa 7321 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag 7381 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg 7441 acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag 7501 cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg 7561 gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat 7621 cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata 7681 tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat 7741 cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag 7801 aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga 7861 taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca 7921 ccgcacagca agcggccggc cgctgatctt cagacctgga ggaggagata tgagggacaa 7981 ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc 8041 caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt 8101 gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac 8161 ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc 8221 tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc 8281 aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg 8341 ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc 8401 tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta 8461 cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca 8521 agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg 8581 gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt 8641 ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca 8701 gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg 8761 agagagagac agagacagat ccattcgatt agtgaacgga tctcgacggt atcgccttta 8821 aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa 8881 cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt 8941 attacaggga cagcagagat ccagtttatc gatgagtaat tcatacaaaa ggactcgccc 9001 ctgccttggg gaatcccagg gaccgtcgtt aaactcccac taacgtagaa cccagagatc 9061 gctgcgttcc cgccccctca cccgcccgct ctcgtcatca ctgaggtgga gaagagcatg 9121 cgtgag //
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