质粒名称: | pUB-Cas9 |
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出品公司: | 爱迪基因 |
目录编号: | 86556 |
存储实验室: | Andreas Weber |
载体骨架基本信息 |
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载体名称: | pKB65 |
载体出品公司: | Kristin Bernhardt |
载体抗性: | 卡那霉素 |
空载体大小: | 11630 bp |
完整质粒大小: | 14121 bp |
载体修饰: | -- |
载体类型: | 植物表达,CRISPR |
筛选标记: | 潮霉素 |
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: | Cas9 |
物种来源: | Synthetic |
插入大小: | 4137 bp |
基因突变: | Cas9 is codon optimized for C. reinhardtii |
GenBank编号 | -- |
启动子 | UBIQUITIN10 |
标签/融合蛋白: | -- |
克隆方法: | Gateway克隆 |
5' 克隆位点: | -- |
3' 克隆位点: | -- |
5' 测序引物 | TTTCGTTCGATCCCAATTTC |
3' 测序引物 | aagaccggcaacaggattc |
文献引用方法 |
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材料和方法部分: | pUB-Cas9 was a gift from Andreas Weber (Addgene plasmid # 86556 ; http://n2t.net/addgene:86556 ; RRID:Addgene_86556) |
参考文献部分: | An Efficient Visual Screen for CRISPR/Cas9 Activity in Arabidopsis thaliana. Hahn F, Mantegazza O, Greiner A, Hegemann P, Eisenhut M, Weber AP. Front Plant Sci. 2017 Jan 24;8:39. doi: 10.3389/fpls.2017.00039. eCollection 2017. 10.3389/fpls.2017.00039 PubMed 28174584 |
买家导航
pUBCas9载体基本信息
pUBCas9载体图谱质粒图谱和多克隆位点信息
pUBCas9载体简介
pUBCas9载体序列
LOCUS pUB-Cas9 14128 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION For ubiquitous expression of Cas9 in planta. sgRNA can be introduced as described in Hahn et al. (2017). ACCESSION . VERSION . KEYWORDS pUB-Cas9 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 14128) AUTHORS Hahn F, Mantegazza O, Greiner A, Hegemann P, Eisenhut M, Weber AP TITLE An Efficient Visual Screen for CRISPR/Cas9 Activity in Arabidopsis thaliana. JOURNAL Front Plant Sci. 2017 Jan 24;8:39. doi: 10.3389/fpls.2017.00039. eCollection 2017. PUBMED 28174584 REFERENCE 2 (bases 1 to 14128) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..14128 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(12..28) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(12..28) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(25..47) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 36..52 /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(60..90) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 105..126 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 317..993 /label=CaMV 35S promoter (enhanced) /note="cauliflower mosaic virus 35S promoter with a duplicated enhancer region" primer_bind 930..950 /label=35S promoter /note="CaMV 35S promoter, forward primer" CDS 1060..2085 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK K" polyA_signal 2126..2300 /label=CaMV poly(A) signal /note="cauliflower mosaic virus polyadenylation signal" misc_feature 2378..2402 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" CDS 2827..3621 /codon_start=1 /gene="aphA-3" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin" /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF" rep_origin 3708..4296 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4197..4216 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4450..4467 /label=L4440 /note="L4440 vector, forward primer" misc_feature 4482..4622 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(4548..4567) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4708..4730 /label=pGEX 3' /note="pGEX vectors, reverse primer" rep_origin 4966..5160 /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" CDS complement(5226..6299) /codon_start=1 /product="replication protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 RepA /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI LVMRYRNLIEGEASAGS" CDS complement(6728..7357) /codon_start=1 /product="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 StaA /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI" misc_feature 8658..8682 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" primer_bind 8870..8892 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 8884..8901 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 8885..8901 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 9610..9634 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 9655..9675 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 9676..13779 /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD ATLIHQSITGLYETRIDLSQLGGD" protein_bind complement(13783..13807) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" terminator 13871..14123 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" ORIGIN 1 ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac 61 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc 121 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg 181 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattggcta gagcagcttg 241 ccaacatggt ggagcacgac actctcgtct actccaagaa tatcaaagat acagtctcag 301 aagaccaaag ggctattgag acttttcaac aaagggtaat atcgggaaac ctcctcggat 361 tccattgccc agctatctgt cacttcatca aaaggacagt agaaaaggaa ggtggcacct 421 acaaatgcca tcattgcgat aaaggaaagg ctatcgttca agatgcctct gccgacagtg 481 gtcccaaaga tggaccccca cccacgagga gcatcgtgga aaaagaagac gttccaacca 541 cgtcttcaaa gcaagtggat tgatgtgaac atggtggagc acgacactct cgtctactcc 601 aagaatatca aagatacagt ctcagaagac caaagggcta ttgagacttt tcaacaaagg 661 gtaatatcgg gaaacctcct cggattccat tgcccagcta tctgtcactt catcaaaagg 721 acagtagaaa aggaaggtgg cacctacaaa tgccatcatt gcgataaagg aaaggctatc 781 gttcaagatg cctctgccga cagtggtccc aaagatggac ccccacccac gaggagcatc 841 gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag tggattgatg tgatatctcc 901 actgacgtaa gggatgacgc acaatcccac tatccttcgc aagacccttc ctctatataa 961 ggaagttcat ttcatttgga gaggacacgc tgaaatcacc agtctctctc tacaaatcta 1021 tctctctcga gctttcgcag atccgggggg caatgagata tgaaaaagcc tgaactcacc 1081 gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag 1141 ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc 1201 ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt 1261 gcatcggccg cgctcccgat tccggaagtg cttgacattg gggagtttag cgagagcctg 1321 acctattgca tctcccgccg ttcacagggt gtcacgttgc aagacctgcc tgaaaccgaa 1381 ctgcccgctg ttctacaacc ggtcgcggag gctatggatg cgatcgctgc ggccgatctt 1441 agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg 1501 cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac 1561 gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac 1621 tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac 1681 aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac 1741 gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc 1801 tacttcgagc ggaggcatcc ggagcttgca ggatcgccac gactccgggc gtatatgctc 1861 cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct 1921 tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca 1981 caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt actcgccgat 2041 agtggaaacc gacgccccag cactcgtccg agggcaaaga aatagagtag atgccgaccg 2101 ggatctgtcg atcgacaagc tcgagtttct ccataataat gtgtgagtag ttcccagata 2161 agggaattag ggttcctata gggtttcgct catgtgttga gcatataaga aacccttagt 2221 atgtatttgt atttgtaaaa tacttctatc aataaaattt ctaattccta aaaccaaaat 2281 ccagtactaa aatccagatc ccccgaatta attcggcgtt aattcagtac attaaaaacg 2341 tccgcaatgt gttattaagt tgtctaagcg tcaatttgtt tacaccacaa tatatcctgc 2401 caccagccag ccaacagctc cccgaccggc agctcggcac aaaatcacca ctcgatacag 2461 gcagcccatc agtccgggac ggcgtcagcg ggagagccgt tgtaaggcgg cagactttgc 2521 tcatgttacc gatgctattc ggaagaacgg caactaagct gccgggtttg aaacacggat 2581 gatctcgcgg agggtagcat gttgattgta acgatgacag agcgttgctg cctgtgatca 2641 ccgcggtttc aaaatcggct ccgtcgatac tatgttatac gccaactttg aaaacaactt 2701 tgaaaaagct gttttctggt atttaaggtt ttagaatgca aggaacagtg aattggagtt 2761 cgtcttgtta taattagctt cttggggtat ctttaaatac tgtagaaaag aggaaggaaa 2821 taataaatgg ctaaaatgag aatatcaccg gaattgaaaa aactgatcga aaaataccgc 2881 tgcgtaaaag atacggaagg aatgtctcct gctaaggtat ataagctggt gggagaaaat 2941 gaaaacctat atttaaaaat gacggacagc cggtataaag ggaccaccta tgatgtggaa 3001 cgggaaaagg acatgatgct atggctggaa ggaaagctgc ctgttccaaa ggtcctgcac 3061 tttgaacggc atgatggctg gagcaatctg ctcatgagtg aggccgatgg cgtcctttgc 3121 tcggaagagt atgaagatga acaaagccct gaaaagatta tcgagctgta tgcggagtgc 3181 atcaggctct ttcactccat cgacatatcg gattgtccct atacgaatag cttagacagc 3241 cgcttagccg aattggatta cttactgaat aacgatctgg ccgatgtgga ttgcgaaaac 3301 tgggaagaag acactccatt taaagatccg cgcgagctgt atgatttttt aaagacggaa 3361 aagcccgaag aggaacttgt cttttcccac ggcgacctgg gagacagcaa catctttgtg 3421 aaagatggca aagtaagtgg ctttattgat cttgggagaa gcggcagggc ggacaagtgg 3481 tatgacattg ccttctgcgt ccggtcgatc agggaggata tcggggaaga acagtatgtc 3541 gagctatttt ttgacttact ggggatcaag cctgattggg agaaaataaa atattatatt 3601 ttactggatg aattgtttta gtacctagaa tgcatgacca aaatccctta acgtgagttt 3661 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt 3721 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt 3781 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag 3841 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta 3901 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat 3961 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg 4021 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg 4081 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac 4141 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga 4201 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt 4261 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta 4321 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat 4381 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg 4441 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc 4501 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct 4561 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg 4621 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat 4681 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt 4741 catcaccgaa acgcgcgagg cagggtgcct tgatgtgggc gccggcggtc gagtggcgac 4801 ggcgcggctt gtccgcgccc tggtagattg cctggccgta ggccagccat ttttgagcgg 4861 ccagcggccg cgataggccg acgcgaagcg gcggggcgta gggagcgcag cgaccgaagg 4921 gtaggcgctt tttgcagctc ttcggctgtg cgctggccag acagttatgc acaggccagg 4981 cgggttttaa gagttttaat aagttttaaa gagttttagg cggaaaaatc gccttttttc 5041 tcttttatat cagtcactta catgtgtgac cggttcccaa tgtacggctt tgggttccca 5101 atgtacgggt tccggttccc aatgtacggc tttgggttcc caatgtacgt gctatccaca 5161 ggaaagagac cttttcgacc tttttcccct gctagggcaa tttgccctag catctgctcc 5221 gtacattagg aaccggcgga tgcttcgccc tcgatcaggt tgcggtagcg catgactagg 5281 atcgggccag cctgccccgc ctcctccttc aaatcgtact ccggcaggtc atttgacccg 5341 atcagcttgc gcacggtgaa acagaacttc ttgaactctc cggcgctgcc actgcgttcg 5401 tagatcgtct tgaacaacca tctggcttct gccttgcctg cggcgcggcg tgccaggcgg 5461 tagagaaaac ggccgatgcc gggatcgatc aaaaagtaat cggggtgaac cgtcagcacg 5521 tccgggttct tgccttctgt gatctcgcgg tacatccaat cagctagctc gatctcgatg 5581 tactccggcc gcccggtttc gctctttacg atcttgtagc ggctaatcaa ggcttcaccc 5641 tcggataccg tcaccaggcg gccgttcttg gccttcttcg tacgctgcat ggcaacgtgc 5701 gtggtgttta accgaatgca ggtttctacc aggtcgtctt tctgctttcc gccatcggct 5761 cgccggcaga acttgagtac gtccgcaacg tgtggacgga acacgcggcc gggcttgtct 5821 cccttccctt cccggtatcg gttcatggat tcggttagat gggaaaccgc catcagtacc 5881 aggtcgtaat cccacacact ggccatgccg gccggccctg cggaaacctc tacgtgcccg 5941 tctggaagct cgtagcggat cacctcgcca gctcgtcggt cacgcttcga cagacggaaa 6001 acggccacgt ccatgatgct gcgactatcg cgggtgccca cgtcatagag catcggaacg 6061 aaaaaatctg gttgctcgtc gcccttgggc ggcttcctaa tcgacggcgc accggctgcc 6121 ggcggttgcc gggattcttt gcggattcga tcagcggccg cttgccacga ttcaccgggg 6181 cgtgcttctg cctcgatgcg ttgccgctgg gcggcctgcg cggccttcaa cttctccacc 6241 aggtcatcac ccagcgccgc gccgatttgt accgggccgg atggtttgcg accgctcacg 6301 ccgattcctc gggcttgggg gttccagtgc cattgcaggg ccggcaggca acccagccgc 6361 ttacgcctgg ccaaccgccc gttcctccac acatggggca ttccacggcg tcggtgcctg 6421 gttgttcttg attttccatg ccgcctcctt tagccgctaa aattcatcta ctcatttatt 6481 catttgctca tttactctgg tagctgcgcg atgtattcag atagcagctc ggtaatggtc 6541 ttgccttggc gtaccgcgta catcttcagc ttggtgtgat cctccgccgg caactgaaag 6601 ttgacccgct tcatggctgg cgtgtctgcc aggctggcca acgttgcagc cttgctgctg 6661 cgtgcgctcg gacggccggc acttagcgtg tttgtgcttt tgctcatttt ctctttacct 6721 cattaactca aatgagtttt gatttaattt cagcggccag cgcctggacc tcgcgggcag 6781 cgtcgccctc gggttctgat tcaagaacgg ttgtgccggc ggcggcagtg cctgggtagc 6841 tcacgcgctg cgtgatacgg gactcaagaa tgggcagctc gtacccggcc agcgcctcgg 6901 caacctcacc gccgatgcgc gtgcctttga tcgcccgcga cacgacaaag gccgcttgta 6961 gccttccatc cgtgacctca atgcgctgct taaccagctc caccaggtcg gcggtggccc 7021 atatgtcgta agggcttggc tgcaccggaa tcagcacgaa gtcggctgcc ttgatcgcgg 7081 acacagccaa gtccgccgcc tggggcgctc cgtcgatcac tacgaagtcg cgccggccga 7141 tggccttcac gtcgcggtca atcgtcgggc ggtcgatgcc gacaacggtt agcggttgat 7201 cttcccgcac ggccgcccaa tcgcgggcac tgccctgggg atcggaatcg actaacagaa 7261 catcggcccc ggcgagttgc agggcgcggg ctagatgggt tgcgatggtc gtcttgcctg 7321 acccgccttt ctggttaagt acagcgataa ccttcatgcg ttccccttgc gtatttgttt 7381 atttactcat cgcatcatat acgcagcgac cgcatgacgc aagctgtttt actcaaatac 7441 acatcacctt tttagacggc ggcgctcggt ttcttcagcg gccaagctgg ccggccaggc 7501 cgccagcttg gcatcagaca aaccggccag gatttcatgc agccgcacgg ttgagacgtg 7561 cgcgggcggc tcgaacacgt acccggccgc gatcatctcc gcctcgatct cttcggtaat 7621 gaaaaacggt tcgtcctggc cgtcctggtg cggtttcatg cttgttcctc ttggcgttca 7681 ttctcggcgg ccgccagggc gtcggcctcg gtcaatgcgt cctcacggaa ggcaccgcgc 7741 cgcctggcct cggtgggcgt cacttcctcg ctgcgctcaa gtgcgcggta cagggtcgag 7801 cgatgcacgc caagcagtgc agccgcctct ttcacggtgc ggccttcctg gtcgatcagc 7861 tcgcgggcgt gcgcgatctg tgccggggtg agggtagggc gggggccaaa cttcacgcct 7921 cgggccttgg cggcctcgcg cccgctccgg gtgcggtcga tgattaggga acgctcgaac 7981 tcggcaatgc cggcgaacac ggtcaacacc atgcggccgg ccggcgtggt ggtgtcggcc 8041 cacggctctg ccaggctacg caggcccgcg ccggcctcct ggatgcgctc ggcaatgtcc 8101 agtaggtcgc gggtgctgcg ggccaggcgg tctagcctgg tcactgtcac aacgtcgcca 8161 gggcgtaggt ggtcaagcat cctggccagc tccgggcggt cgcgcctggt gccggtgatc 8221 ttctcggaaa acagcttggt gcagccggcc gcgtgcagtt cggcccgttg gttggtcaag 8281 tcctggtcgt cggtgctgac gcgggcatag cccagcaggc cagcggcggc gctcttgttc 8341 atggcgtaat gtctccggtt ctagtcgcaa gtattctact ttatgcgact aaaacacgcg 8401 acaagaaaac gccaggaaaa gggcagggcg gcagcctgtc gcgtaactta ggacttgtgc 8461 gacatgtcgt tttcagaaga cggctgcact gaacgtcaga agccgactgc actatagcag 8521 cggaggggtt ggatcaaagt actttgatcc cgaggggaac cctgtggttg gcatgcacat 8581 acaaatggac gaacggataa accttttcac gcccttttaa atatccgatt attctaataa 8641 acgctctttt ctcttaggtt tacccgccaa tatatcctgt caaacactga tagtttaaac 8701 tgaaggcggg aaacgacaat ctgatccaag ctcaagctgc tctagcattc gccattcagg 8761 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg 8821 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga 8881 cgttgtaaaa cgacggccag tgccaagctt gattacgaat tcgagctcgg tacccgacga 8941 gtcagtaata aacggcgtca aagtggttgc agccggcaca cacgagtcgt gtttatcaac 9001 tcaaagcaca aatacttttc ctcaacctaa aaataaggca attagccaaa aacaactttg 9061 cgtgtaaaca acgctcaata cacgtgtcat tttattatta gctattgctt caccgcctta 9121 gctttctcgt gacctagtcg tcctcgtctt ttcttcttct tcttctataa aacaataccc 9181 aaagagctct tcttcttcac aattcagatt tcaatttctc aaaatcttaa aaactttctc 9241 tcaattctct ctaccgtgat caaggtaaat ttctgtgttc cttattctct caaaatcttc 9301 gattttgttt tcgttcgatc ccaatttcgt atatgttctt tggtttagat tctgttaatc 9361 ttagatcgaa gacgattttc tgggtttgat cgttagatat catcttaatt ctcgattagg 9421 gtttcataga tatcatccga tttgttcaaa taatttgagt tttgtcgaat aattactctt 9481 cgatttgtga tttctatcta gatctggtgt tagtttctag tttgtgcgat cgaatttgta 9541 gattaatctg agtttttctg attaacagct cgagtgcggg ccccccctcg aggcgcgcca 9601 agctatcaaa caagtttgta caaaaaagca ggctaccata tggccagcag cccccccaag 9661 aagaagcgca aggtcgacaa gaagtactcc atcggcctgg acatcggcac gaacagcgtg 9721 ggctgggccg tgatcaccga cgagtacaag gtgccgtcca agaagttcaa ggtgctgggc 9781 aacaccgacc gccacagcat caagaagaac ctgatcggcg ccctgctgtt cgactcgggc 9841 gagaccgccg aggcgacgcg cctgaagcgc accgcgcgcc gccgctacac gcgccgcaag 9901 aaccgcatct gctacctgca ggagatcttc agcaacgaga tggccaaggt ggacgactcg 9961 ttcttccacc gcctggagga gtccttcctg gtggaggagg acaagaagca cgagcgccac 10021 cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc gacgatctac 10081 cacctgcgca agaagctggt ggacagcacc gacaaggcgg acctgcgcct gatctacctg 10141 gccctggcgc acatgatcaa gttccgcggc cacttcctga tcgagggcga cctgaacccc 10201 gacaactcgg acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc 10261 gaggagaacc cgatcaacgc ctcgggcgtg gacgccaagg cgatcctgtc cgcgcgcctg 10321 tccaagagcc gccgcttgga gaacctgatc gcccagctgc ccggcgagaa gaagaacggc 10381 ctgttcggca acctgatcgc gctgagcctg ggcctgacgc cgaacttcaa gtcgaacttc 10441 gacctggccg aggacgcgaa gctgcagctg agcaaggaca cctacgacga cgacctggac 10501 aacctgctgg cccagatcgg cgaccagtac gcggacctgt tcctggccgc gaagaacctg 10561 tcggacgcca tcctgctgtc cgacatcctg cgcgtgaaca ccgagatcac gaaggccccc 10621 ctgtcggcgt ccatgatcaa gcgctacgac gagcaccacc aggacctgac cctgctgaag 10681 gcgctggtgc gccagcagct gccggagaag tacaaggaga tcttcttcga ccagtccaag 10741 aacggctacg ccggctacat cgacggcggc gccagccagg aggagttcta caagttcatc 10801 aagcccatcc tggagaagat ggacggcacg gaggagctgc tggtgaagct gaaccgcgag 10861 gacctgctgc gcaagcagcg caccttcgac aacggcagca tcccccacca gatccacctg 10921 ggcgagctgc acgccatcct gcgccgccag gaggacttct acccgttcct gaaggacaac 10981 cgcgagaaga tcgagaagat cctgacgttc cgcatcccct actacgtggg cccgctggct 11041 cgcggcaact cccgcttcgc gtggatgacc cgcaagagcg aggagaccat cacgccctgg 11101 aacttcgagg aggtggtgga caagggcgcc agcgcgcagt cgttcatcga gcgcatgacc 11161 aacttcgaca agaacctgcc caacgagaag gtgctgccga agcactccct gctgtacgag 11221 tacttcaccg tgtacaacga gctgacgaag gtgaagtacg tgaccgaggg catgcgcaag 11281 cccgccttcc tgagcggcga gcagaagaag gcgatcgtgg acctgctgtt caagaccaac 11341 cgcaaggtga cggtgaagca gctgaaggag gactacttca agaagatcga gtgcttcgac 11401 agcgtggaga tctcgggcgt ggaggaccgc ttcaacgcca gcctgggcac ctaccacgac 11461 ctgctgaaga tcatcaagga caaggacttc ctggacaacg aggagaacga ggacatcctg 11521 gaggacatcg tgctgaccct gacgctgttc gaggaccgcg agatgatcga ggagcgcctg 11581 aagacgtacg cccacctgtt cgacgacaag gtgatgaagc agctgaagcg ccgccgctac 11641 accggctggg gccgcctgag ccgcaagctg atcaacggca tccgcgacaa gcagtcgggc 11701 aagaccatcc tggacttcct gaagtccgac ggcttcgcga accgcaactt catgcagctg 11761 atccacgacg actcgctgac cttcaaggag gacatccaga aggcccaggt gtcgggccag 11821 ggcgactccc tgcacgagca catcgccaac ctggcgggct cccccgcgat caagaagggc 11881 atcctgcaga ccgtgaaggt ggtggacgag ctggtgaagg tgatgggccg ccacaagccg 11941 gagaacatcg tgatcgagat ggcccgcgag aaccagacca cgcagaaggg ccagaagaac 12001 agccgcgagc gcatgaagcg catcgaggag ggcatcaagg agctgggctc gcagatcctg 12061 aaggagcacc ccgtggagaa cacccagctg cagaacgaga agctgtacct gtactacctg 12121 cagaacggcc gcgacatgta cgtggaccag gagctggaca tcaaccgcct gtccgactac 12181 gacgtggacc acatcgtgcc ccagagcttc ctgaaggacg actcgatcga caacaaggtg 12241 ctgacccgca gcgacaagaa ccgcggcaag agcgacaacg tgccgtcgga ggaggtggtg 12301 aagaagatga agaactactg gcgccagctg ctgaacgcca agctgatcac gcagcgcaag 12361 ttcgacaacc tgaccaaggc cgagcgcggc ggcctgtcgg agctggacaa ggcgggcttc 12421 atcaagcgcc agctggtgga gacccgccag atcacgaagc acgtggcgca gatcctggac 12481 tcccgcatga acacgaagta cgacgagaac gacaagctga tccgcgaggt gaaggtgatc 12541 accctgaagt ccaagctggt cagcgacttc cgcaaggact tccagttcta caaggtgcgc 12601 gagatcaaca actaccacca cgcccacgac gcgtacctga acgccgtggt gggcaccgcg 12661 ctgatcaaga agtaccccaa gctggagagc gagttcgtgt acggcgacta caaggtgtac 12721 gacgtgcgca agatgatcgc caagtcggag caggagatcg gcaaggccac cgcgaagtac 12781 ttcttctact ccaacatcat gaacttcttc aagaccgaga tcacgctggc caacggcgag 12841 atccgcaagc gccccctgat cgagaccaac ggcgagacgg gcgagatcgt gtgggacaag 12901 ggccgcgact tcgcgaccgt gcgcaaggtg ctgagcatgc cccaggtgaa catcgtgaag 12961 aagaccgagg tgcagacggg cggcttctcc aaggagagca tcctgccgaa gcgcaactcg 13021 gacaagctga tcgcccgcaa gaaggactgg gaccccaaga agtacggcgg cttcgactcc 13081 ccgaccgtgg cctacagcgt gctggtggtg gcgaaggtgg agaagggcaa gtccaagaag 13141 ctgaagagcg tgaaggagct gctgggcatc accatcatgg agcgcagctc gttcgagaag 13201 aaccccatcg acttcctgga ggcgaagggc tacaaggagg tgaagaagga cctgatcatc 13261 aagctgccga agtactcgct gttcgagctg gagaacggcc gcaagcgcat gctggcctcc 13321 gcgggcgagc tgcagaaggg caacgagctg gccctgccca gcaagtacgt gaacttcctg 13381 tacctggcgt cccactacga gaagctgaag ggctcgccgg aggacaacga gcagaagcag 13441 ctgttcgtgg agcagcacaa gcactacctg gacgagatca tcgagcagat ctcggagttc 13501 tccaagcgcg tgatcctggc cgacgcgaac ctggacaagg tgctgagcgc ctacaacaag 13561 caccgcgaca agcccatccg cgagcaggcg gagaacatca tccacctgtt caccctgacg 13621 aacctgggcg ccccggccgc gttcaagtac ttcgacacca cgatcgaccg caagcgctac 13681 accagcacga aggaggtgct ggacgcgacc ctgatccacc agagcatcac cggcctgtac 13741 gagacgcgca tcgacctgtc gcagctgggc ggcgactaat gtacccagct ttcttgtaca 13801 aagtggttcg ataattctta attaactagt tctagagcgg ccgccaccgc ggtggagctc 13861 gaatttcccc gatcgttcaa acatttggca ataaagtttc ttaagattga atcctgttgc 13921 cggtcttgcg atgattatca tataatttct gttgaattac gttaagcatg taataattaa 13981 catgtaatgc atgacgttat ttatgagatg ggtttttatg attagagtcc cgcaattata 14041 catttaatac gcgatagaaa acaaaatata gcgcgcaaac taggataaat tatcgcgcgc 14101 ggtgtcatct atgttactag atcgggaa //