质粒名称: | pLVCT-tTR-KRAB |
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出品公司: | 爱迪基因 |
目录编号: | 11643 |
存储实验室: | Patrick Aebischer, Didier Trono |
载体骨架基本信息 |
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载体名称: | -- |
载体出品公司: | -- |
载体抗性: | 氨苄青霉素 |
空载体大小: | 12877 bp |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | 哺乳动物表达,慢病毒 |
筛选标记: | -- |
高拷贝/低拷贝: | 高拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | CAG promoters, GFP, tTR-KRAB, Tet-on |
物种来源: | -- |
插入大小: | -- |
基因突变: | -- |
GenBank编号 | -- |
启动子 | -- |
标签/融合蛋白: | -- |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | -- |
3' 克隆位点: | -- |
5' 测序引物 | See map |
3' 测序引物 | -- |
文献引用方法 |
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材料和方法部分: | pLVCT-tTR-KRAB was a gift from Patrick Aebischer & Didier Trono (Addgene plasmid # 11643 ; http://n2t.net/addgene:11643 ; RRID:Addgene_11643) |
参考文献部分: | A versatile tool for conditional gene expression and knockdown. Szulc J, Wiznerowicz M, Sauvain MO, Trono D, Aebischer P. Nat Methods. 2006 Feb . 3(2):109-16. 10.1038/nmeth846 PubMed 16432520 |
买家导航
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 pLVCT-tTR-KRAB 12884 bp ds-DNA circular SYN 23-MAR-2020 DEFINITION Tet-regulated (Tet-on) lentiviral vector for transgene (CAG promoter) - OR - shRNA (H1 promoter when subcloned from pLVTHM (Addgene#12247)) - 2nd generation. ACCESSION . VERSION . KEYWORDS pLVCT-tTR-KRAB SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12884) AUTHORS Szulc J, Wiznerowicz M, Sauvain MO, Trono D, Aebischer P TITLE A versatile tool for conditional gene expression and knockdown. JOURNAL Nat Methods. 2006 Feb . 3(2):109-16. PUBMED 16432520 REFERENCE 2 (bases 1 to 12884) AUTHORS . TITLE Direct Submission JOURNAL Exported Mar 23, 2020 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..12884 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 6..639 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 686..811 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1304..1537 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." enhancer 2061..2440 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2442..2719 /label=chicken beta-actin promoter intron 2720..3736 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 3660..3682 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 3744..3763 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" misc_feature 3822..3939 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" regulatory 4016..4025 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 4022..4738 /codon_start=1 /product="enhanced GFP" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(4067..4088) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4328..4347) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4675..4696 /label=EGFP-C /note="EGFP, forward primer" misc_feature 4834..5385 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4978..4995) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 5205..5224 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5388..6005 /codon_start=1 /gene="tetR from transposon Tn10" /product="tetracycline repressor TetR" /label=TetR /note="TetR binds to the tetracycline operator tetO to inhibit transcription. This inhibition can be relieved by adding tetracycline or doxycycline." /translation="MARLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESG" primer_bind complement(5461..5480) /label=Tet-R /note="Tetracycline resistance gene, reverse primer" CDS 6009..6029 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 6132..6326 /codon_start=1 /gene="ZNF10" /gene_synonym="KOX1" /product="Kruppel-associated box (KRAB) transcriptional repression domain from the human zinc finger protein ZNF10 (Margolin et al., 1994)" /label=KRAB /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY QLTKPDVILRLEKGEEPWLV" misc_feature 6469..7057 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(6522..6542) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(6940..6951) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" protein_bind 7195..7465 /label=tetracycline response element /bound_moiety="tetracycline repressor TetR" /note="contains seven copies of the tetracycline operator tetO" LTR 7504..7684 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" promoter complement(7709..7727) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(7710..7727) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(8017..8036) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind complement(8250..8269) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 8369..8391 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(8429..8447) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 8515..8619 /gene="bla" /label=AmpR promoter CDS 8620..9480 /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 complement(8838..8857) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 9651..10239 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 10140..10159 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 10393..10410 /label=L4440 /note="L4440 vector, forward primer" primer_bind complement(10480..10500) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 10485..10814 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 10665..10800 /label=SV40 ori /note="SV40 origin of replication" primer_bind 10727..10746 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" intron 11948..12013 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS 12143..12163 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 12588..12722 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(12625..12644) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 12679..12698 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" ORIGIN 1 gtagttggaa gggctaattc actcccaaag aagacaagat atccttgatc tgtggatcta 61 ccacacacaa ggctacttcc ctgattagca gaactacaca ccagggccag gggtcagata 121 tccactgacc tttggatggt gctacaagct agtaccagtt gagccagata aggtagaaga 181 ggccaataaa ggagagaaca ccagcttgtt acaccctgtg agcctgcatg ggatggatga 241 cccggagaga gaagtgttag agtggaggtt tgacagccgc ctagcatttc atcacgtggc 301 ccgagagctg catccggagt acttcaagaa ctgctgatat cgagcttgct acaagggact 361 ttccgctggg gactttccag ggaggcgtgg cctgggcggg actggggagt ggcgagccct 421 cagatcctgc atataagcag ctgctttttg cctgtactgg gtctctctgg ttagaccaga 481 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct 541 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat 601 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt 661 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg 721 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg 781 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg 841 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt 901 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga 961 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact 1021 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa 1081 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc 1141 acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa 1201 gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg 1261 caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg 1321 ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg 1381 ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg 1441 cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc 1501 tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa 1561 aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac 1621 agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct 1681 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat 1741 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt 1801 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg 1861 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc 1921 tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag 1981 acagagacag atccattcga ttagtgaacg gatctcgacg gtatcgatca cgagactagc 2041 ctcgaccatc gatgtcgatc gacattgatt attgactagt tattaatagt aatcaattac 2101 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg 2161 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc 2221 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac 2281 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa 2341 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac 2401 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt 2461 ctgcttcact ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt 2521 ttaattattt tgtgcagcga tgggggcggg gggggggggg gggcgcgcgc naggsggggc 2581 ggggcggggc gaggggcggg gcggggcgag gcggagaggt gcggcggcag ccaatcagag 2641 cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa 2701 gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc 2761 cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg 2821 gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc 2881 ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag ggccctttgt gcggggggag 2941 cggctcgggg ggtgcgtgcg tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct 3001 gcccggcggc tgtgagcgct gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg 3061 cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa 3121 aggctgcgtg cggggtgtgt gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg 3181 gctgcaaccc cccctgcacc cccctccccg agttgctgag cacggcccgg cttcgggtgc 3241 ggggctccgt acggggcgtg gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt 3301 gggggtgccg ggcggggcgg ggccgcctcg ggccggggag ggctcggggg aggggcgcgg 3361 cggcccccgg agcgccggcg gctgtcgagg cgcggcgagc cgcagccatt gccttttatg 3421 gtaatcgtgc gagagggcgc agggacttcc tttgtcccaa atctgtgcgg agccgaaatc 3481 tgggaggcgc cgccgcaccc cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag 3541 gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct 3601 ccagcctcgg ggctgtccgc ggggggacgg ctgccttcgg gggggacggg gcagggcggg 3661 gttcggcttc tggcgtgtga ccggcggctc tagagcctct gctaaccatg ttcatgcctt 3721 cttctttttc ctacagctcc tgggcaacgt gctggttatt gtgctgtctc atcattttgg 3781 caaagaatta aatttaatta atctcgacgg tatcggttaa cttttaaaag aaaagggggg 3841 attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact 3901 aaagaattac aaaaacaaat tacaaaaatt caaaatttta tcgatcacga gactagcctc 3961 gaggtttaaa ctacgggatc caggcctaag cttacgcgtc ctagcgctac cggtcgccac 4021 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga 4081 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta 4141 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac 4201 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa 4261 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt 4321 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct 4381 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca 4441 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa 4501 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc 4561 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca 4621 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt 4681 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagtc 4741 cggactcaga tctcgactag ctagtagcta gctagctagt cgagctcaag cttcgaattc 4801 ccgggctgca ggaattccgc cccccccccc ctaacgttac tggccgaagc cgcttggaat 4861 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg 4921 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc 4981 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt 5041 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg 5101 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac 5161 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg 5221 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg 5281 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaacgtc taggcccccc 5341 gaaccacggg gacgtggttt tcctttgaaa aacacgatga taataccatg gctagattag 5401 ataaaagtaa agtgattaac agcgcattag agctgcttaa tgaggtcgga atcgaaggtt 5461 taacaacccg taaactcgcc cagaagctag gtgtagagca gcctacattg tattggcatg 5521 taaaaaataa gcgggctttg ctcgacgcct tagccattga gatgttagat aggcaccata 5581 ctcacttttg ccctttagaa ggggaaagct ggcaagattt tttacgtaat aacgctaaaa 5641 gttttagatg tgctttacta agtcatcgcg atggagcaaa agtacattta ggtacacggc 5701 ctacagaaaa acagtatgaa actctcgaaa atcaattagc ctttttatgc caacaaggtt 5761 tttcactaga gaatgcatta tatgcactca gcgctgtggg gcattttact ttaggttgcg 5821 tattggaaga tcaagagcat caagtcgcta aagaagaaag ggaaacacct actactgata 5881 gtatgccgcc attattacga caagctatcg aattatttga tcaccaaggt gcagagccag 5941 ccttcttatt cggccttgaa ttgatcatat gcggattaga aaaacaactt aaatgtgaaa 6001 gtgggtcgcc aaaaaagaag agaaaggtcg acggcggtgg tgctttgtct cctcagcact 6061 ctgctgtcac tcaaggaagt atcatcaaga acaaggaggg catggatgct aagtcactaa 6121 ctgcctggtc ccggacactg gtgaccttca aggatgtatt tgtggacttc accagggagg 6181 agtggaagct gctggacact gctcagcaga tcgtgtacag aaatgtgatg ctggagaact 6241 ataagaacct ggtttccttg ggttatcagc ttactaagcc agatgtgatc ctccggttgg 6301 agaagggaga agagccctgg ctggtggaga gagaaattca ccaagagacc catcctgatt 6361 cagagactgc atttgaaatc aaatcatcag tttaaggatc tcgactagct agtagctagc 6421 tagctagtcg agctcaagct tcgaattcga tgggactagt catatgataa tcaacctctg 6481 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta 6541 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt 6601 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc 6661 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt 6721 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg 6781 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac 6841 aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc 6901 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac 6961 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct 7021 cagacgagtc ggatctccct ttgggccgcc tccccgcatc ggtacgtatg gccaggtacc 7081 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg 7141 gggactggaa gggctaattc actcccaacg aagacaagat ctcgagttta ccactcccta 7201 tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt 7261 gaaagtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac 7321 tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag 7381 aaaagtgaaa gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt 7441 taccactccc tatcagtgat agagaaaagt gaaagtcgga tcgatctgct ttttgcttgt 7501 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac 7561 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg 7621 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct 7681 agcagcatct agaattaatt ccgtgtattc tatagtgtca cctaaatcgt atgtgtatga 7741 tacataaggt tatgtattaa ttgtagccgc gttctaacga caatatgtac aagcctaatt 7801 gtgtagcatc tggcttactg aagcagaccc tatcatctct ctcgtaaact gccgtcagag 7861 tcggtttggt tggacgaacc ttctgagttt ctggtaacgc cgtcccgcac ccggaaatgg 7921 tcagcgaacc aatcagcagg gtcatcgcta gccagatcct ctacgccgga cgcatcgtgg 7981 ccggcatcac cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg 8041 gggaagatcg ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg 8101 caggccccgt ggccggggga ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg 8161 cggcggtgct caacggcctc aacctactac tgggctgctt cctaatgcag gagtcgcata 8221 agggagagcg tcgaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc 8281 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca 8341 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg 8401 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat 8461 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga 8521 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa 8581 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt 8641 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg 8701 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg 8761 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg 8821 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag 8881 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca 8941 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg 9001 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc 9061 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg 9121 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg 9181 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac 9241 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg 9301 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg 9361 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact 9421 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa 9481 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt 9541 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag 9601 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct 9661 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt 9721 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg 9781 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct 9841 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc 9901 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg 9961 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa 10021 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg 10081 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg 10141 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga 10201 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt 10261 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct 10321 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga 10381 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg 10441 cctctccccg cgcgttggcc gattcattaa tgcagctgtg gaatgtgtgt cagttagggt 10501 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 10561 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 10621 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc 10681 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg 10741 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc 10801 taggcttttg caaaaagctt ggacacaaga caggcttgcg agatatgttt gagaatacca 10861 ctttatcccg cgtcagggag aggcagtgcg taaaaagacg cggactcatg tgaaatactg 10921 gtttttagtg cgccagatct ctataatctc gcgcaaccta ttttcccctc gaacactttt 10981 taagccgtag ataaacaggc tgggacactt cacatgagcg aaaaatacat cgtcacctgg 11041 gacatgttgc agatccatgc acgtaaactc gcaagccgac tgatgccttc tgaacaatgg 11101 aaaggcatta ttgccgtaag ccgtggcggt ctgtaccggg tgcgttactg gcgcgtgaac 11161 tgggtattcg tcatgtcgat accgtttgta tttccagcta cgatcacgac aaccagcgcg 11221 agcttaaagt gctgaaacgc gcagaaggcg atggcgaagg cttcatcgtt attgatgacc 11281 tggtggatac cggtggtact gcggttgcga ttcgtgaaat gtatccaaaa gcgcactttg 11341 tcaccatctt cgcaaaaccg gctggtcgtc cgctggttga tgactatgtt gttgatatcc 11401 cgcaagatac ctggattgaa cagccgtggg atatgggcgt cgtattcgtc ccgccaatct 11461 ccggtcgcta atcttttcaa cgcctggcac tgccgggcgt tgttcttttt aacttcaggc 11521 gggttacaat agtttccagt aagtattctg gaggctgcat ccatgacaca ggcaaacctg 11581 agcgaaaccc tgttcaaacc ccgctttaaa catcctgaaa cctcgacgct agtccgccgc 11641 tttaatcacg gcgcacaacc gcctgtgcag tcggcccttg atggtaaaac catccctcac 11701 tggtatcgca tgattaaccg tctgatgtgg atctggcgcg gcattgaccc acgcgaaatc 11761 ctcgacgtcc aggcacgtat tgtgatgagc gatgccgaac gtaccgacga tgatttatac 11821 gatacggtga ttggctaccg tggcggcaac tggatttatg agtgggcccc ggatctttgt 11881 gaaggaacct tacttctgtg gtgtgacata attggacaaa ctacctacag agatttaaag 11941 ctctaaggta aatataaaat ttttaagtgt ataatgtgtt aaactactga ttctaattgt 12001 ttgtgtattt tagattccaa cctatggaac tgatgaatgg gagcagtggt ggaatgcctt 12061 taatgaggaa aacctgtttt gctcagaaga aatgccatct agtgatgatg aggctactgc 12121 tgactctcaa cattctactc ctccaaaaaa gaagagaaag gtagaagacc ccaaggactt 12181 tccttcagaa ttgctaagtt ttttgagtca tgctgtgttt agtaatagaa ctcttgcttg 12241 ctttgctatt tacaccacaa aggaaaaagc tgcactgcta tacaagaaaa ttatggaaaa 12301 atattctgta acctttataa gtaggcataa cagttataat cataacatac tgttttttct 12361 tactccacac aggcatagag tgtctgctat taataactat gctcaaaaat tgtgtacctt 12421 tagcttttta atttgtaaag gggttaataa ggaatatttg atgtatagtg ccttgactag 12481 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac 12541 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg 12601 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt 12661 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga 12721 tcaactggat aactcaagct aaccaaaatc atcccaaact tcccacccca taccctatta 12781 ccactgccaa ttacctgtgg tttcatttac tctaaacctg tgattcctct gaattatttt 12841 cattttaaag aaattgtatt tgttaaatat gtactacaaa ctta //