质粒名称: | pMJ806 |
---|---|
出品公司: | 爱迪基因 |
目录编号: | 39312 |
存储实验室: | Jennifer Doudna |
载体骨架基本信息 |
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载体名称: | pEC-K-MBP |
载体出品公司: | pET-based custom vector |
载体抗性: | 卡那霉素25ug/mL |
空载体大小: | 6400 bp |
完整质粒大小: | 10500 bp |
载体修饰: | -- |
载体类型: | 细菌表达,CRISPR |
筛选标记: | -- |
高拷贝/低拷贝: | 低拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | Cas9,Csn1 |
物种来源: | Streptococcus pyogenes |
插入大小: | 4104 bp |
基因突变: | -- |
GenBank编号 | AAK33936.1 13622193 |
启动子 | T7 |
标签/融合蛋白: |
MBP(主干上的N终端)
His6(主干上的N终端)
|
克隆方法: | Ligation Independent克隆 |
5' 克隆位点: | -- |
3' 克隆位点: | -- |
5' 测序引物 | MBP-forward |
3' 测序引物 | T7 terminator |
文献引用方法 |
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材料和方法部分: | pMJ806 was a gift from Jennifer Doudna (Addgene plasmid # 39312 ; http://n2t.net/addgene:39312 ; RRID:Addgene_39312) |
参考文献部分: | A Programmable Dual-RNA-Guided DNA Endonuclease in Adaptive Bacterial Immunity. Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier E. Science. 2012 Jun 28. 10.1126/science.1225829 PubMed 22745249 |
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 pMJ806 10577 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pMJ806 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10577) AUTHORS Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier E TITLE A Programmable Dual-RNA-Guided DNA Endonuclease in Adaptive Bacterial Immunity. JOURNAL Science. 2012 Jun 28. PUBMED 22745249 REFERENCE 2 (bases 1 to 10577) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10577 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 5..144 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(70..89) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 230..252 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(246..437) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(1246..2328) /codon_start=1 /gene="lacI" /product="lac repressor" /label=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)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" primer_bind 2290..2309 /label=LacI-R /note="LacI, reverse primer" promoter complement(2329..2406) /gene="lacI" /label=lacI promoter primer_bind 2612..2631 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind 2715..2734 /label=T7 /note="T7 promoter, forward primer" promoter 2715..2733 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 2734..2758 /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)." RBS 2773..2795 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 2806..2823 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 2827..3924 /codon_start=1 /gene="malE (mutated)" /product="maltose binding protein from E. coli" /label=MBP /note="This version of the gene does not encode a signal sequence, so MBP will remain in the cytosol." /translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW YAVRTAVINAASGRQTVDEALKDAQT" primer_bind 3898..3921 /label=MBP-F /note="Maltose binding protein, forward primer" CDS 3955..3975 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 3985..8091 /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 8156..8173 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(8226..8244) /label=T7 Term /note="T7 terminator, reverse primer" terminator 8240..8287 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 8324..8779 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(8411..8430) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 8621..8642 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(8871..9686) /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind 9594..9613 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 9678..9697 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 9808..10396 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 10297..10316 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 10550..10567 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg 61 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg 121 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga 181 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc 241 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca 301 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg 361 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt 421 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg 481 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg 541 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa 601 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc 661 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt 721 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac 781 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca 841 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc 901 cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga 961 ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg 1021 atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc 1081 acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg 1141 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat 1201 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc 1261 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc 1321 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg 1381 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc 1441 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc 1501 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat 1561 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat 1621 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc 1681 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg 1741 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa 1801 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt 1861 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc 1921 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg 1981 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga 2041 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga 2101 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc 2161 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt 2221 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata 2281 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct 2341 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat 2401 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta 2461 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc 2521 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga 2581 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa 2641 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct 2701 cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata acaattcccc 2761 tctagaaata attttgttta actttaagaa ggagatatac atatgcacca tcaccatcac 2821 catggaaaaa tcgaagaagg taaactggta atctggatta acggcgataa aggctataac 2881 ggtctcgctg aagtcggtaa gaaattcgag aaagataccg gaattaaagt caccgttgag 2941 catccggata aactggaaga gaaattccca caggttgcgg caactggcga tggccctgac 3001 attatcttct gggcacacga ccgctttggt ggctacgctc aatctggcct gttggctgaa 3061 atcaccccgg acaaagcgtt ccaggacaag ctgtatccgt ttacctggga tgccgtacgt 3121 tacaacggca agctgattgc ttacccgatc gctgttgaag cgttatcgct gatttataac 3181 aaagatctgc tgccgaaccc gccaaaaacc tgggaagaga tcccggcgct ggataaagaa 3241 ctgaaagcga aaggtaagag cgcgctgatg ttcaacctgc aagaaccgta cttcacctgg 3301 ccgctgattg ctgctgacgg gggttatgcg ttcaagtatg aaaacggcaa gtacgacatt 3361 aaagacgtgg gcgtggataa cgctggcgcg aaagcgggtc tgaccttcct ggttgacctg 3421 attaaaaaca aacacatgaa tgcagacacc gattactcca tcgcagaagc tgcctttaat 3481 aaaggcgaaa cagcgatgac catcaacggc ccgtgggcat ggtccaacat cgacaccagc 3541 aaagtgaatt atggtgtaac ggtactgccg accttcaagg gtcaaccatc caaaccgttc 3601 gttggcgtgc tgagcgcagg tattaacgcc gccagtccga acaaagagct ggcaaaagag 3661 ttcctcgaaa actatctgct gactgatgaa ggtctggaag cggttaataa agacaaaccg 3721 ctgggtgccg tagcgctgaa gtcttacgag gaagagttgg cgaaagatcc acgtattgcc 3781 gccactatgg aaaacgccca gaaaggtgaa atcatgccga acatcccgca gatgtccgct 3841 ttctggtatg ccgtgcgtac tgcggtgatc aacgccgcca gcggtcgtca gactgtcgat 3901 gaagccctga aagacgcgca gactaattcg agctcgaaca acaacaacac tagtgaaaac 3961 ctgtatttcc agggagcagc ctcgatggat aagaaatact caataggctt agatatcggc 4021 acaaatagcg tcggatgggc ggtgatcact gatgaatata aggttccgtc taaaaagttc 4081 aaggttctgg gaaatacaga ccgccacagt atcaaaaaaa atcttatagg ggctctttta 4141 tttgacagtg gagagacagc ggaagcgact cgtctcaaac ggacagctcg tagaaggtat 4201 acacgtcgga agaatcgtat ttgttatcta caggagattt tttcaaatga gatggcgaaa 4261 gtagatgata gtttctttca tcgacttgaa gagtcttttt tggtggaaga agacaagaag 4321 catgaacgtc atcctatttt tggaaatata gtagatgaag ttgcttatca tgagaaatat 4381 ccaactatct atcatctgcg aaaaaaattg gtagattcta ctgataaagc ggatttgcgc 4441 ttaatctatt tggccttagc gcatatgatt aagtttcgtg gtcatttttt gattgaggga 4501 gatttaaatc ctgataatag tgatgtggac aaactattta tccagttggt acaaacctac 4561 aatcaattat ttgaagaaaa ccctattaac gcaagtggag tagatgctaa agcgattctt 4621 tctgcacgat tgagtaaatc aagacgatta gaaaatctca ttgctcagct ccccggtgag 4681 aagaaaaatg gcttatttgg gaatctcatt gctttgtcat tgggtttgac ccctaatttt 4741 aaatcaaatt ttgatttggc agaagatgct aaattacagc tttcaaaaga tacttacgat 4801 gatgatttag ataatttatt ggcgcaaatt ggagatcaat atgctgattt gtttttggca 4861 gctaagaatt tatcagatgc tattttactt tcagatatcc taagagtaaa tactgaaata 4921 actaaggctc ccctatcagc ttcaatgatt aaacgctacg atgaacatca tcaagacttg 4981 actcttttaa aagctttagt tcgacaacaa cttccagaaa agtataaaga aatctttttt 5041 gatcaatcaa aaaacggata tgcaggttat attgatgggg gagctagcca agaagaattt 5101 tataaattta tcaaaccaat tttagaaaaa atggatggta ctgaggaatt attggtgaaa 5161 ctaaatcgtg aagatttgct gcgcaagcaa cggacctttg acaacggctc tattccccat 5221 caaattcact tgggtgagct gcatgctatt ttgagaagac aagaagactt ttatccattt 5281 ttaaaagaca atcgtgagaa gattgaaaaa atcttgactt ttcgaattcc ttattatgtt 5341 ggtccattgg cgcgtggcaa tagtcgtttt gcatggatga ctcggaagtc tgaagaaaca 5401 attaccccat ggaattttga agaagttgtc gataaaggtg cttcagctca atcatttatt 5461 gaacgcatga caaactttga taaaaatctt ccaaatgaaa aagtactacc aaaacatagt 5521 ttgctttatg agtattttac ggtttataac gaattgacaa aggtcaaata tgttactgaa 5581 ggaatgcgaa aaccagcatt tctttcaggt gaacagaaga aagccattgt tgatttactc 5641 ttcaaaacaa atcgaaaagt aaccgttaag caattaaaag aagattattt caaaaaaata 5701 gaatgttttg atagtgttga aatttcagga gttgaagata gatttaatgc ttcattaggt 5761 acctaccatg atttgctaaa aattattaaa gataaagatt ttttggataa tgaagaaaat 5821 gaagatatct tagaggatat tgttttaaca ttgaccttat ttgaagatag ggagatgatt 5881 gaggaaagac ttaaaacata tgctcacctc tttgatgata aggtgatgaa acagcttaaa 5941 cgtcgccgtt atactggttg gggacgtttg tctcgaaaat tgattaatgg tattagggat 6001 aagcaatctg gcaaaacaat attagatttt ttgaaatcag atggttttgc caatcgcaat 6061 tttatgcagc tgatccatga tgatagtttg acatttaaag aagacattca aaaagcacaa 6121 gtgtctggac aaggcgatag tttacatgaa catattgcaa atttagctgg tagccctgct 6181 attaaaaaag gtattttaca gactgtaaaa gttgttgatg aattggtcaa agtaatgggg 6241 cggcataagc cagaaaatat cgttattgaa atggcacgtg aaaatcagac aactcaaaag 6301 ggccagaaaa attcgcgaga gcgtatgaaa cgaatcgaag aaggtatcaa agaattagga 6361 agtcagattc ttaaagagca tcctgttgaa aatactcaat tgcaaaatga aaagctctat 6421 ctctattatc tccaaaatgg aagagacatg tatgtggacc aagaattaga tattaatcgt 6481 ttaagtgatt atgatgtcga tcacattgtt ccacaaagtt tccttaaaga cgattcaata 6541 gacaataagg tcttaacgcg ttctgataaa aatcgtggta aatcggataa cgttccaagt 6601 gaagaagtag tcaaaaagat gaaaaactat tggagacaac ttctaaacgc caagttaatc 6661 actcaacgta agtttgataa tttaacgaaa gctgaacgtg gaggtttgag tgaacttgat 6721 aaagctggtt ttatcaaacg ccaattggtt gaaactcgcc aaatcactaa gcatgtggca 6781 caaattttgg atagtcgcat gaatactaaa tacgatgaaa atgataaact tattcgagag 6841 gttaaagtga ttaccttaaa atctaaatta gtttctgact tccgaaaaga tttccaattc 6901 tataaagtac gtgagattaa caattaccat catgcccatg atgcgtatct aaatgccgtc 6961 gttggaactg ctttgattaa gaaatatcca aaacttgaat cggagtttgt ctatggtgat 7021 tataaagttt atgatgttcg taaaatgatt gctaagtctg agcaagaaat aggcaaagca 7081 accgcaaaat atttctttta ctctaatatc atgaacttct tcaaaacaga aattacactt 7141 gcaaatggag agattcgcaa acgccctcta atcgaaacta atggggaaac tggagaaatt 7201 gtctgggata aagggcgaga ttttgccaca gtgcgcaaag tattgtccat gccccaagtc 7261 aatattgtca agaaaacaga agtacagaca ggcggattct ccaaggagtc aattttacca 7321 aaaagaaatt cggacaagct tattgctcgt aaaaaagact gggatccaaa aaaatatggt 7381 ggttttgata gtccaacggt agcttattca gtcctagtgg ttgctaaggt ggaaaaaggg 7441 aaatcgaaga agttaaaatc cgttaaagag ttactaggga tcacaattat ggaaagaagt 7501 tcctttgaaa aaaatccgat tgacttttta gaagctaaag gatataagga agttaaaaaa 7561 gacttaatca ttaaactacc taaatatagt ctttttgagt tagaaaacgg tcgtaaacgg 7621 atgctggcta gtgccggaga attacaaaaa ggaaatgagc tggctctgcc aagcaaatat 7681 gtgaattttt tatatttagc tagtcattat gaaaagttga agggtagtcc agaagataac 7741 gaacaaaaac aattgtttgt ggagcagcat aagcattatt tagatgagat tattgagcaa 7801 atcagtgaat tttctaagcg tgttatttta gcagatgcca atttagataa agttcttagt 7861 gcatataaca aacatagaga caaaccaata cgtgaacaag cagaaaatat tattcattta 7921 tttacgttga cgaatcttgg agctcccgct gcttttaaat attttgatac aacaattgat 7981 cgtaaacgat atacgtctac aaaagaagtt ttagatgcca ctcttatcca tcaatccatc 8041 actggtcttt atgaaacacg cattgatttg agtcagctag gaggtgacta acgaggccgg 8101 tgctttgcag gatccgaatt cgagctccgt cgacaagctt gcggccgcac tcgagcacca 8161 ccaccaccac cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc 8221 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg 8281 ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc 8341 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc 8401 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc 8461 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc 8521 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg 8581 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact 8641 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt 8701 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa 8761 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt 8821 gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc ttagaaaaac 8881 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt 8941 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca 9001 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc 9061 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt 9121 gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc 9181 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg 9241 agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg 9301 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat 9361 acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta 9421 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc 9481 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc 9541 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga 9601 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa 9661 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac 9721 agttttattg ttcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga 9781 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg 9841 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc 9901 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct 9961 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc 10021 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt 10081 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg 10141 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct 10201 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag 10261 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag 10321 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg 10381 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg 10441 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac 10501 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt 10561 gagcgaggaa gcggaag //