质粒名称: | pX330a dCas9-KRAB |
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出品公司: | 爱迪基因 |
目录编号: | 92361 |
存储实验室: | Tatjana Sauka-Spengler |
载体骨架基本信息 |
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载体名称: | x330 |
载体出品公司: | -- |
载体抗性: | 氨苄青霉素 |
空载体大小: | 4332 bp |
完整质粒大小: | 8731 bp |
载体修饰: | -- |
载体类型: | 哺乳细胞表达,CRISPR |
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: | dCas9-KRAB |
物种来源: | -- |
插入大小: | 4399 bp |
基因突变: | -- |
GenBank编号 | -- |
启动子 | CAG |
标签/融合蛋白: | -- |
克隆方法: | Gibson克隆 |
5' 克隆位点: | -- |
3' 克隆位点: | -- |
5' 测序引物 | tctgactgaccgcgttactc |
3' 测序引物 | aggaaaggacagtgggagtg |
文献引用方法 |
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材料和方法部分: | pX330a dCas9-KRAB was a gift from Tatjana Sauka-Spengler (Addgene plasmid # 92361 ; http://n2t.net/addgene:92361 ; RRID:Addgene_92361) |
参考文献部分: | Genome and epigenome engineering CRISPR toolkit for in vivo modulation of cis-regulatory interactions and gene expression in the chicken embryo. Williams RM, Senanayake U, Artibani M, Taylor G, Wells D, Ahmed AA, Sauka-Spengler T. Development. 2018 Feb 23;145(4). pii: dev.160333. doi: 10.1242/dev.160333. 10.1242/dev.160333 PubMed 29386245 |
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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 pX330a dCas9-KRAB 8731 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Modified CAG promoter-containing vector for ubiquitous expression of catalytically inactive Cas9 fused to KRAB repressor domain. For targeted enhancer inactivation in chicken embryos.. ACCESSION . VERSION . KEYWORDS pX330a dCas9-KRAB SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8731) AUTHORS Williams RM, Senanayake U, Artibani M, Taylor G, Wells D, Ahmed AA, Sauka-Spengler T TITLE Genome and epigenome engineering CRISPR toolkit for probing in vivo cis-regulatory interactions in the chicken embryo JOURNAL BioRxiv REFERENCE 2 (bases 1 to 8731) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8731 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 24..612 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 513..532 /label=pBR322ori-F /note="pBR322 origin, forward primer" promoter 674..914 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 674..694 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 845..864 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_RNA 941..1016 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" enhancer 1113..1398 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer; contains an 18-bp deletion relative to the standard CMV enhancer" promoter 1400..1677 /label=chicken beta-actin promoter intron 1678..1906 /label=hybrid intron /note="hybrid between chicken beta-actin (CBA) and minute virus of mice (MMV) introns (Gray et al., 2011)" CDS 1927..1947 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1957..6060 /codon_start=1 /product="catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9m4 /note="RNA-guided DNA-binding protein that lacks endonuclease activity due to the mutations D10A, D839A, H840A, and N863A (Mali et al., 2013)" /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVAAIVPQSFLKDDSIDNKVLTRSDKARGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" CDS 6073..6093 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 6127..6321 /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="RTLVTFKDVFVDFTREEWKLLDTAQQILYRNVMLENYKNLVSLGY QLTKPDVILRLEKGEEPWLV" CDS 6388..6408 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" primer_bind complement(6445..6462) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 6451..6658 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 6667..6807 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 6667..6796 /label=AAV2 ITR rep_origin 6882..7337 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6969..6988) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7179..7200 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(7354..7373) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 7473..7495 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(7533..7551) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 7619..7723 /gene="bla" /label=AmpR promoter CDS 7724..8584 /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(7942..7961) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" ORIGIN 1 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg 61 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact 121 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg 181 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg 241 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac 301 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca 361 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga 421 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc 481 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct 541 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg 601 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct 661 tttgctcaca tgtgagggcc tatttcccat gattccttca tatttgcata tacgatacaa 721 ggctgttaga gagataattg gaattaattt gactgtaaac acaaagatat tagtacaaaa 781 tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa 841 aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat 901 cttgtggaaa ggacgaaaca ccgggtcttc gagaagacct gttttagagc tagaaatagc 961 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt 1021 ttgttttaga gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc 1081 caattctgca gacaaatggc tctagaggta cccgttacat aacttacggt aaatggcccg 1141 cctggctgac cgcccaacga cccccgccca ttgacgtcaa tagtaacgcc aatagggact 1201 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa 1261 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg 1321 cattgtgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta 1381 gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc 1441 cccccctccc cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg 1501 ggggcggggg gggggggggg gcgcgcgcca ggcggggcgg ggcggggcga ggggcggggc 1561 ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct 1621 tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga 1681 gtcgctgcga cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc 1741 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc 1801 gggctgtaat tagctgagca agaggtaagg gtttaaggga tggttggttg gtggggtatt 1861 aatgtttaat tacctggagc acctgcctga aatcactttt tttcaggttg gaccggtgcc 1921 accatgccca agaagaagag gaaggtggga agggggatgg acaagaagta ctccattggg 1981 ctcgctatcg gcacaaacag cgtcggctgg gccgtcatta cggacgagta caaggtgccg 2041 agcaaaaaat tcaaagttct gggcaatacc gatcgccaca gcataaagaa gaacctcatt 2101 ggcgccctcc tgttcgactc cggggagacg gccgaagcca cgcggctcaa aagaacagca 2161 cggcgcagat atacccgcag aaagaatcgg atctgctacc tgcaggagat ctttagtaat 2221 gagatggcta aggtggatga ctctttcttc cataggctgg aggagtcctt tttggtggag 2281 gaggataaaa agcacgagcg ccacccaatc tttggcaata tcgtggacga ggtggcgtac 2341 catgaaaagt acccaaccat atatcatctg aggaagaagc ttgtagacag tactgataag 2401 gctgacttgc ggttgatcta tctcgcgctg gcgcatatga tcaaatttcg gggacacttc 2461 ctcatcgagg gggacctgaa cccagacaac agcgatgtcg acaaactctt tatccaactg 2521 gttcagactt acaatcagct tttcgaagag aacccgatca acgcatccgg agttgacgcc 2581 aaagcaatcc tgagcgctag gctgtccaaa tcccggcggc tcgaaaacct catcgcacag 2641 ctccctgggg agaagaagaa cggcctgttt ggtaatctta tcgccctgtc actcgggctg 2701 acccccaact ttaaatctaa cttcgacctg gccgaagatg ccaagcttca actgagcaaa 2761 gacacctacg atgatgatct cgacaatctg ctggcccaga tcggcgacca gtacgcagac 2821 ctttttttgg cggcaaagaa cctgtcagac gccattctgc tgagtgatat tctgcgagtg 2881 aacacggaga tcaccaaagc tccgctgagc gctagtatga tcaagcgcta tgatgagcac 2941 caccaagact tgactttgct gaaggccctt gtcagacagc aactgcctga gaagtacaag 3001 gaaattttct tcgatcagtc taaaaatggc tacgccggat acattgacgg cggagcaagc 3061 caggaggaat tttacaaatt tattaagccc atcttggaaa aaatggacgg caccgaggag 3121 ctgctggtaa agcttaacag agaagatctg ttgcgcaaac agcgcacttt cgacaatgga 3181 agcatccccc accagattca cctgggcgaa ctgcacgcta tcctcaggcg gcaagaggat 3241 ttctacccct ttttgaaaga taacagggaa aagattgaga aaatcctcac atttcggata 3301 ccctactatg taggccccct cgcccgggga aattccagat tcgcgtggat gactcgcaaa 3361 tcagaagaga ccatcactcc ctggaacttc gaggaagtcg tggataaggg ggcctctgcc 3421 cagtccttca tcgaaaggat gactaacttt gataaaaatc tgcctaacga aaaggtgctt 3481 cctaaacact ctctgctgta cgagtacttc acagtttata acgagctcac caaggtcaaa 3541 tacgtcacag aagggatgag aaagccagca ttcctgtctg gagagcagaa gaaagctatc 3601 gtggacctcc tcttcaagac gaaccggaaa gttaccgtga aacagctcaa agaggactat 3661 ttcaaaaaga ttgaatgttt cgactctgtt gaaatcagcg gagtggagga tcgcttcaac 3721 gcatccctgg gaacgtatca cgatctcctg aaaatcatta aagacaagga cttcctggac 3781 aatgaggaga acgaggacat tcttgaggac attgtcctca cccttacgtt gtttgaagat 3841 agggagatga ttgaagaacg cttgaaaact tacgctcatc tcttcgacga caaagtcatg 3901 aaacagctca agaggcgccg atatacagga tgggggcggc tgtcaagaaa actgatcaat 3961 gggatccgag acaagcagag tggaaagaca atcctggatt ttcttaagtc cgatggattt 4021 gccaaccgga acttcatgca gttgatccat gatgactctc tcacctttaa ggaggacatc 4081 cagaaagcac aagtttctgg ccagggggac agtctccacg agcacatcgc taatcttgca 4141 ggtagcccag ctatcaaaaa gggaatactg cagaccgtta aggtcgtgga tgaactcgtc 4201 aaagtaatgg gaaggcataa gcccgagaat atcgttatcg agatggcccg agagaaccaa 4261 actacccaga agggacagaa gaacagtagg gaaaggatga agaggattga agagggtata 4321 aaagaactgg ggtcccaaat ccttaaggaa cacccagttg aaaacaccca gcttcagaat 4381 gagaagctct acctgtacta cctgcagaac ggcagggaca tgtacgtgga tcaggaactg 4441 gacatcaatc ggctctccga ctacgacgtg gctgctatcg tgccccagtc ttttctcaaa 4501 gatgattcta ttgataataa agtgttgaca agatccgata aagctagagg gaagagtgat 4561 aacgtcccct cagaagaagt tgtcaagaaa atgaaaaatt attggcggca gctgctgaac 4621 gccaaactga tcacacaacg gaagttcgat aatctgacta aggctgaacg aggtggcctg 4681 tctgagttgg ataaagccgg cttcatcaaa aggcagcttg ttgagacacg ccagatcacc 4741 aagcacgtgg cccaaattct cgattcacgc atgaacacca agtacgatga aaatgacaaa 4801 ctgattcgag aggtgaaagt tattactctg aagtctaagc tggtctcaga tttcagaaag 4861 gactttcagt tttataaggt gagagagatc aacaattacc accatgcgca tgatgcctac 4921 ctgaatgcag tggtaggcac tgcacttatc aaaaaatatc ccaagcttga atctgaattt 4981 gtttacggag actataaagt gtacgatgtt aggaaaatga tcgcaaagtc tgagcaggaa 5041 ataggcaagg ccaccgctaa gtacttcttt tacagcaata ttatgaattt tttcaagacc 5101 gagattacac tggccaatgg agagattcgg aagcgaccac ttatcgaaac aaacggagaa 5161 acaggagaaa tcgtgtggga caagggtagg gatttcgcga cagtccggaa ggtcctgtcc 5221 atgccgcagg tgaacatcgt taaaaagacc gaagtacaga ccggaggctt ctccaaggaa 5281 agtatcctcc cgaaaaggaa cagcgacaag ctgatcgcac gcaaaaaaga ttgggacccc 5341 aagaaatacg gcggattcga ttctcctaca gtcgcttaca gtgtactggt tgtggccaaa 5401 gtggagaaag ggaagtctaa aaaactcaaa agcgtcaagg aactgctggg catcacaatc 5461 atggagcgat caagcttcga aaaaaacccc atcgactttc tcgaggcgaa aggatataaa 5521 gaggtcaaaa aagacctcat cattaagctt cccaagtact ctctctttga gcttgaaaac 5581 ggccggaaac gaatgctcgc tagtgcgggc gagctgcaga aaggtaacga gctggcactg 5641 ccctctaaat acgttaattt cttgtatctg gccagccact atgaaaagct caaagggtct 5701 cccgaagata atgagcagaa gcagctgttc gtggaacaac acaaacacta ccttgatgag 5761 atcatcgagc aaataagcga attctccaaa agagtgatcc tcgccgacgc taacctcgat 5821 aaggtgcttt ctgcttacaa taagcacagg gataagccca tcagggagca ggcagaaaac 5881 attatccact tgtttactct gaccaacttg ggcgcgcctg cagccttcaa gtacttcgac 5941 accaccatag acagaaagcg gtacacctct acaaaggagg tcctggacgc cacactgatt 6001 catcagtcaa ttacggggct ctatgaaaca agaatcgacc tctctcagct cggtggagac 6061 agcagggctg accccaagaa gaagaggaag gtggctagcg atgctaagtc actgactgcc 6121 tggtcccgga cactggtgac cttcaaggat gtgtttgtgg acttcaccag ggaggagtgg 6181 aagctgctgg acactgctca gcagatcctg tacagaaatg tgatgctgga gaactataag 6241 aacctggttt ccttgggtta tcagcttact aagccagatg tgatcctccg gttggagaag 6301 ggagaagagc cctggctggt ggagagagaa attcaccaag agacccatcc tgattcagag 6361 actgcatttg aaatcaaatc atcagttccg aaaaagaaac gcaaagttgc tagctgataa 6421 gaattcctag agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg 6481 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct 6541 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg 6601 gggtggggca ggacagcaag ggggaggatt gggaagagaa tagcaggcat gctggggagc 6661 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca 6721 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga 6781 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc 6841 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc 6901 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct 6961 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg 7021 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga 7081 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt 7141 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg 7201 aacaacactc aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc 7261 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat 7321 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt 7381 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc 7441 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc 7501 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt 7561 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg 7621 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca 7681 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt 7741 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga 7801 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga 7861 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat 7921 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca 7981 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt 8041 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac 8101 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct 8161 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga 8221 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac 8281 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat 8341 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg 8401 ctggtttatt gctgataaat ctggagccgg tgagcgtgga agccgcggta tcattgcagc 8461 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc 8521 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg 8581 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta 8641 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg 8701 tgagttttcg ttccactgag cgtcagaccc c //