质粒名称: | LentiCRISPRv2GFP |
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出品公司: | 爱迪基因 |
目录编号: | 82416 |
存储实验室: | David Feldser |
载体骨架基本信息 |
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载体名称: | lentiCRISPR v2 |
载体出品公司: | Feng Zhang |
载体抗性: | 氨苄西林 |
完整质粒大小: | 13129bp |
载体修饰: | 在gfp中克隆以取代puro电阻 |
载体类型: | 慢病毒,crispr |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | GFP |
备用名称:
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EGFP |
物种来源: | 维多利亚多管发光水母Aequorea victoria |
插入大小: | 720bp |
启动子 | EFS (P2A) |
克隆方法: | 吉布森克隆 |
5' 测序引物 | ATGGTGTCTAAGGGCGAAGA |
文献引用方法 |
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材料和方法部分: | |
参考文献部分: | Systematic in vivo inactivation of chromatin regulating enzymes identifies Setd2 as a potent tumor suppressor in lung adenocarcinoma. Walter DM, Venancio OS, Buza EL, Tobias JW, Deshpande C, Gudiel AA, Kim-Kiselak C, Cicchini M, Yates TJ, Feldser DM. Cancer Res. 2017 Feb 15. pii: canres.2159.2016. doi: 10.1158/0008-5472.CAN-16-2159. 10.1158/0008-5472.CAN-16-2159 PubMed 28202515 |
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 LentiCRISPRv2GFP 13131 bp ds-DNA circular SYN 12-MAR-2019 DEFINITION Lentiviral vector expressing GFP alongside Cas9 and an sgRNA cloning site. ACCESSION . VERSION . KEYWORDS LentiCRISPRv2GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 13131) AUTHORS Walter DM, Venancio OS, Buza EL, Tobias JW, Deshpande C, Gudiel AA, Kim-Kiselak C, Cicchini M, Yates TJ, Feldser DM TITLE Systematic in vivo inactivation of chromatin regulating enzymes identifies Setd2 as a potent tumor suppressor in lung adenocarcinoma. JOURNAL Cancer Res. 2017 Feb 15. pii: canres.2159.2016. doi: 10.1158/0008-5472.CAN-16-2159. PUBMED 28202515 REFERENCE 2 (bases 1 to 13131) AUTHORS . TITLE Direct Submission JOURNAL Exported Mar 12, 2019 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..13131 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..241 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 1..21 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 172..191 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_RNA 266..341 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" promoter 403..614 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" regulatory 633..642 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 639..4742 /codon_start=1 /product="Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9 /note="generates RNA-guided double strand breaks in DNA" /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" CDS 4743..4790 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" CDS 4791..4814 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 4824..4880 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" primer_bind complement(4926..4947) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(5187..5206) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 5534..5555 /label=EGFP-C /note="EGFP, forward primer" misc_feature 5616..6204 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5669..5689) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(6087..6098) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 6276..6509 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" primer_bind complement(6535..6552) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 6541..6765 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 6811..7239 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6898..6917) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7108..7129 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(7248..7268) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 7253..7582 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 7433..7568 /label=SV40 ori /note="SV40 origin of replication" primer_bind 7495..7514 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 7630..7677 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 7696..8070 /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" polyA_signal 8200..8321 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(8237..8256) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 8291..8310 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(8370..8386) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8370..8386) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(8383..8405) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 8394..8410 /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(8418..8448) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8463..8484 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(8601..8618) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(8772..9360) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(8852..8871) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(9531..10391) /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 10154..10173 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(10392..10496) /gene="bla" /label=AmpR promoter primer_bind complement(10571..10590) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 10762..11141 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 11143..11341 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 11296..11316 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 11359..11539 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 11586..11711 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 12204..12437 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." misc_feature 12964..13081 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" ORIGIN 1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag 61 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga 121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat 181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga 241 cgaaacaccg gagacggacg tctctgtttt agagctagaa atagcaagtt aaaataaggc 301 tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cttttttgaa ttcgctagct 361 aggtcttgaa aggagtggga attggctccg gtgcccgtca gtgggcagag cgcacatcgc 421 ccacagtccc cgagaagttg gggggagggg tcggcaattg atccggtgcc tagagaaggt 481 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg 541 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg 601 ccgccagaac acaggaccgg ttctagagcg ctgccaccat ggacaagaag tacagcatcg 661 gcctggacat cggcaccaac tctgtgggct gggccgtgat caccgacgag tacaaggtgc 721 ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca cagcatcaag aagaacctga 781 tcggagccct gctgttcgac agcggcgaaa cagccgaggc cacccggctg aagagaaccg 841 ccagaagaag atacaccaga cggaagaacc ggatctgcta tctgcaagag atcttcagca 901 acgagatggc caaggtggac gacagcttct tccacagact ggaagagtcc ttcctggtgg 961 aagaggataa gaagcacgag cggcacccca tcttcggcaa catcgtggac gaggtggcct 1021 accacgagaa gtaccccacc atctaccacc tgagaaagaa actggtggac agcaccgaca 1081 aggccgacct gcggctgatc tatctggccc tggcccacat gatcaagttc cggggccact 1141 tcctgatcga gggcgacctg aaccccgaca acagcgacgt ggacaagctg ttcatccagc 1201 tggtgcagac ctacaaccag ctgttcgagg aaaaccccat caacgccagc ggcgtggacg 1261 ccaaggccat cctgtctgcc agactgagca agagcagacg gctggaaaat ctgatcgccc 1321 agctgcccgg cgagaagaag aatggcctgt tcggaaacct gattgccctg agcctgggcc 1381 tgacccccaa cttcaagagc aacttcgacc tggccgagga tgccaaactg cagctgagca 1441 aggacaccta cgacgacgac ctggacaacc tgctggccca gatcggcgac cagtacgccg 1501 acctgtttct ggccgccaag aacctgtccg acgccatcct gctgagcgac atcctgagag 1561 tgaacaccga gatcaccaag gcccccctga gcgcctctat gatcaagaga tacgacgagc 1621 accaccagga cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca 1681 aagagatttt cttcgaccag agcaagaacg gctacgccgg ctacattgac ggcggagcca 1741 gccaggaaga gttctacaag ttcatcaagc ccatcctgga aaagatggac ggcaccgagg 1801 aactgctcgt gaagctgaac agagaggacc tgctgcggaa gcagcggacc ttcgacaacg 1861 gcagcatccc ccaccagatc cacctgggag agctgcacgc cattctgcgg cggcaggaag 1921 atttttaccc attcctgaag gacaaccggg aaaagatcga gaagatcctg accttccgca 1981 tcccctacta cgtgggccct ctggccaggg gaaacagcag attcgcctgg atgaccagaa 2041 agagcgagga aaccatcacc ccctggaact tcgaggaagt ggtggacaag ggcgcttccg 2101 cccagagctt catcgagcgg atgaccaact tcgataagaa cctgcccaac gagaaggtgc 2161 tgcccaagca cagcctgctg tacgagtact tcaccgtgta taacgagctg accaaagtga 2221 aatacgtgac cgagggaatg agaaagcccg ccttcctgag cggcgagcag aaaaaggcca 2281 tcgtggacct gctgttcaag accaaccgga aagtgaccgt gaagcagctg aaagaggact 2341 acttcaagaa aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa gatcggttca 2401 acgcctccct gggcacatac cacgatctgc tgaaaattat caaggacaag gacttcctgg 2461 acaatgagga aaacgaggac attctggaag atatcgtgct gaccctgaca ctgtttgagg 2521 acagagagat gatcgaggaa cggctgaaaa cctatgccca cctgttcgac gacaaagtga 2581 tgaagcagct gaagcggcgg agatacaccg gctggggcag gctgagccgg aagctgatca 2641 acggcatccg ggacaagcag tccggcaaga caatcctgga tttcctgaag tccgacggct 2701 tcgccaacag aaacttcatg cagctgatcc acgacgacag cctgaccttt aaagaggaca 2761 tccagaaagc ccaggtgtcc ggccagggcg atagcctgca cgagcacatt gccaatctgg 2821 ccggcagccc cgccattaag aagggcatcc tgcagacagt gaaggtggtg gacgagctcg 2881 tgaaagtgat gggccggcac aagcccgaga acatcgtgat cgaaatggcc agagagaacc 2941 agaccaccca gaagggacag aagaacagcc gcgagagaat gaagcggatc gaagagggca 3001 tcaaagagct gggcagccag atcctgaaag aacaccccgt ggaaaacacc cagctgcaga 3061 acgagaagct gtacctgtac tacctgcaga atgggcggga tatgtacgtg gaccaggaac 3121 tggacatcaa ccggctgtcc gactacgatg tggaccatat cgtgcctcag agctttctga 3181 aggacgactc catcgacaac aaggtgctga ccagaagcga caagaaccgg ggcaagagcg 3241 acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa ctactggcgg cagctgctga 3301 acgccaagct gattacccag agaaagttcg acaatctgac caaggccgag agaggcggcc 3361 tgagcgaact ggataaggcc ggcttcatca agagacagct ggtggaaacc cggcagatca 3421 caaagcacgt ggcacagatc ctggactccc ggatgaacac taagtacgac gagaatgaca 3481 agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga 3541 aggatttcca gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct 3601 acctgaacgc cgtcgtggga accgccctga tcaaaaagta ccctaagctg gaaagcgagt 3661 tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat gatcgccaag agcgagcagg 3721 aaatcggcaa ggctaccgcc aagtacttct tctacagcaa catcatgaac tttttcaaga 3781 ccgagattac cctggccaac ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg 3841 aaaccgggga gatcgtgtgg gataagggcc gggattttgc caccgtgcgg aaagtgctga 3901 gcatgcccca agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag 3961 agtctatcct gcccaagagg aacagcgata agctgatcgc cagaaagaag gactgggacc 4021 ctaagaagta cggcggcttc gacagcccca ccgtggccta ttctgtgctg gtggtggcca 4081 aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa agagctgctg gggatcacca 4141 tcatggaaag aagcagcttc gagaagaatc ccatcgactt tctggaagcc aagggctaca 4201 aagaagtgaa aaaggacctg atcatcaagc tgcctaagta ctccctgttc gagctggaaa 4261 acggccggaa gagaatgctg gcctctgccg gcgaactgca gaagggaaac gaactggccc 4321 tgccctccaa atatgtgaac ttcctgtacc tggccagcca ctatgagaag ctgaagggct 4381 cccccgagga taatgagcag aaacagctgt ttgtggaaca gcacaagcac tacctggacg 4441 agatcatcga gcagatcagc gagttctcca agagagtgat cctggccgac gctaatctgg 4501 acaaagtgct gtccgcctac aacaagcacc gggataagcc catcagagag caggccgaga 4561 atatcatcca cctgtttacc ctgaccaatc tgggagcccc tgccgccttc aagtactttg 4621 acaccaccat cgaccggaag aggtacacca gcaccaaaga ggtgctggac gccaccctga 4681 tccaccagag catcaccggc ctgtacgaga cacggatcga cctgtctcag ctgggaggcg 4741 acaagcgacc tgccgccaca aagaaggctg gacaggctaa gaagaagaaa gattacaaag 4801 acgatgacga taagggatcc ggcgcaacaa acttctctct gctgaaacaa gccggagatg 4861 tcgaagagaa tcctggaccg atggtgtcta agggcgaaga gctgttcacc ggggtggtgc 4921 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg 4981 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc 5041 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc 5101 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg 5161 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga 5221 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg 5281 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca 5341 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg 5401 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg 5461 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg 5521 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca 5581 tggacgagct gtacaagtaa acgcgttaag tcgacaatca acctctggat tacaaaattt 5641 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg 5701 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt 5761 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg 5821 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc 5881 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg 5941 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt 6001 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc 6061 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg 6121 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga 6181 tctccctttg ggccgcctcc ccgcgtcgac tttaagacca atgacttaca aggcagctgt 6241 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg 6301 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg 6361 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt 6421 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc 6481 cttttagtca gtgtggaaaa tctctagcag ggcccgttta aacccgctga tcagcctcga 6541 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc 6601 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc 6661 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt 6721 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa 6781 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg 6841 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc 6901 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa 6961 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac 7021 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt 7081 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca 7141 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt 7201 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca 7261 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct 7321 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca 7381 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc 7441 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta 7501 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt 7561 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga 7621 tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag 7681 gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac 7741 gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag 7801 gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac 7861 caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac 7921 gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc 7981 gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc 8041 gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc 8101 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc 8161 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat 8221 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg 8281 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg 8341 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat 8401 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc 8461 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga 8521 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 8581 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 8641 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 8701 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 8761 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 8821 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 8881 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 8941 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag 9001 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 9061 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 9121 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg 9181 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg 9241 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct 9301 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa 9361 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 9421 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa 9481 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc 9541 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga 9601 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca 9661 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc 9721 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat 9781 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc 9841 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt 9901 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc 9961 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg 10021 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt 10081 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg 10141 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga 10201 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg 10261 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg 10321 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt 10381 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc 10441 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca 10501 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt 10561 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt 10621 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct 10681 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg 10741 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta 10801 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 10861 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 10921 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 10981 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 11041 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 11101 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 11161 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 11221 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 11281 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 11341 gcgcgttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct 11401 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta 11461 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca 11521 gtgtggaaaa tctctagcag tggcgcccga acagggactt gaaagcgaaa gggaaaccag 11581 aggagctctc tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc 11641 ggcgactggt gagtacgcca aaaattttga ctagcggagg ctagaaggag agagatgggt 11701 gcgagagcgt cagtattaag cgggggagaa ttagatcgcg atgggaaaaa attcggttaa 11761 ggccaggggg aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag 11821 aacgattcgc agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg 11881 gacagctaca accatccctt cagacaggat cagaagaact tagatcatta tataatacag 11941 tagcaaccct ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag 12001 acaagataga ggaagagcaa aacaaaagta agaccaccgc acagcaagcg gccgctgatc 12061 ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa 12121 gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag 12181 agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga 12241 agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt 12301 atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa 12361 ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta 12421 aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct 12481 gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc 12541 tggatggagt gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa 12601 gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca 12661 agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg 12721 atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga 12781 gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc 12841 gacaggcccg aaggaataga agaagaaggt ggagagagag acagagacag atccattcga 12901 ttagtgaacg gatcggcact gcgtgcgcca attctgcaga caaatggcag tattcatcca 12961 caattttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat 13021 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt 13081 tcgggtttat tacagggaca gcagagatcc agtttggtta attaaggtac c //