质粒名称: | pJBEI-6409 |
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出品公司: | 爱迪基因 |
目录编号: | 47048 |
存储实验室: | Taek Soon Lee |
载体骨架基本信息 |
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载体名称: | pBbA5c |
载体出品公司: | -- |
载体抗性: | 氯霉素 |
空载体大小: | 3500 bp |
完整质粒大小: | 14932 bp |
载体修饰: | -- |
载体类型: | 细菌表达,合成生物学;布格瑞克 |
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: | Codon-optimized sequences for MEV pathway expression in E. coli to produce limonene |
物种来源: | S. cerevisiae (budding yeast); Mentha spicata, Abies grandis and Staphylococcus aureus |
插入大小: | 11500 bp |
基因突变: | -- |
GenBank编号 | -- |
启动子 | PlacUV5 |
标签/融合蛋白: | -- |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | EcoRI, BglII (未破坏) |
3' 克隆位点: | BamHI, XhoI (未破坏) |
5' 测序引物 | F5 |
3' 测序引物 | R1 |
文献引用方法 |
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材料和方法部分: | pJBEI-6409 was a gift from Taek Soon Lee (Addgene plasmid # 47048 ; http://n2t.net/addgene:47048 ; RRID:Addgene_47048) |
参考文献部分: | Metabolic engineering of Escherichia coli for limonene and perillyl alcohol production. Alonso-Gutierrez J, Chan R, Batth TS, Adams PD, Keasling JD, Petzold CJ, Lee TS. Metab Eng. 2013 May 29;19C:33-41. doi: 10.1016/j.ymben.2013.05.004. 10.1016/j.ymben.2013.05.004 PubMed 23727191 |
买家导航
pJBEI6409载体基本信息
pJBEI6409载体图谱质粒图谱和多克隆位点信息
pJBEI6409载体简介
pJBEI6409载体序列
LOCUS pJBEI-6409 14932 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION BglBrick plasmid (=pBbA5c-MTSAe-T1f-MBI(f)-T1002i-Ptrc-trGPPS(co)-LS) coding for MEV pathway enzymes to produce limonene from glucose in E. coli. ACCESSION . VERSION . KEYWORDS pJBEI-6409 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 14932) AUTHORS Alonso-Gutierrez J, Chan R, Batth TS, Adams PD, Keasling JD, Petzold CJ, Lee TS TITLE Metabolic engineering of Escherichia coli for limonene and perillyl alcohol production. JOURNAL Metab Eng. 2013 May 29;19C:33-41. doi: 10.1016/j.ymben.2013.05.004. PUBMED 23727191 REFERENCE 2 (bases 1 to 14932) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..14932 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(49..1131) /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 1093..1112 /label=LacI-R /note="LacI, reverse primer" promoter complement(1132..1209) /gene="lacI" /label=lacI promoter primer_bind 1415..1434 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" promoter 1520..1550 /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" protein_bind 1558..1574 /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)." CDS complement(2309..2320) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" terminator 5324..5395 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 5411..5438 /label=T7Te terminator /note="phage T7 early transcription terminator" promoter 5650..5679 /label=trc promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" protein_bind 5687..5703 /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)." CDS 8726..8737 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" promoter 10432..10461 /label=trc promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" protein_bind 10469..10485 /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)." terminator 13097..13168 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 13184..13211 /label=T7Te terminator /note="phage T7 early transcription terminator" primer_bind complement(13239..13256) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(13373..13918) /direction=LEFT /label=p15A ori /note="Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin. " primer_bind complement(13453..13472) /label=pBR322ori-F /note="pBR322 origin, forward primer" terminator 14032..14126 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(14147..14806) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind 14721..14740 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" promoter complement(14807..14909) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" ORIGIN 1 gacgtcggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct 61 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 121 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag 181 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg 241 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc 301 ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc cggactcggt 361 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac 421 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc 481 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag 541 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc 601 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact 661 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc 721 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg 781 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat 841 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg 901 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc 961 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac 1021 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg 1081 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac 1141 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc 1201 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca 1261 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg 1321 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca 1381 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag 1441 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga 1501 tcgtttaggc accccaggct ttacacttta tgcttccggc tcgtataatg tgtggaattg 1561 tgagcggata acaatttcag aattcaaaag atcttaggag gaatataaaa tgaaaaattg 1621 tgtcatcgtc agtgcggtac gtactgctat cggtagtttt aacggttcac tcgcttccac 1681 cagcgccatc gacctggggg cgacagtaat taaagccgcc attgaacgtg caaaaatcga 1741 ttcacaacac gttgatgaag tgattatggg taacgtgtta caagccgggc tggggcaaaa 1801 tccggcgcgt caggcactgt taaaaagcgg gctggcagaa acggtgtgcg gattcacggt 1861 caataaagta tgtggttcgg gtcttaaaag tgtggcgctt gccgcccagg ccattcaggc 1921 aggtcaggcg cagagcattg tggcgggggg tatggaaaat atgagtttag ccccctactt 1981 actcgatgca aaagcacgct ctggttatcg tcttggagac ggacaggttt atgacgtaat 2041 cctgcgcgat ggcctgatgt gcgccaccca tggttatcat atggggatta ccgccgaaaa 2101 cgtggctaaa gagtacggaa ttacccgtga aatgcaggat gaactggcgc tacattcaca 2161 gcgtaaagcg gcagccgcaa ttgagtccgg tgcttttaca gccgaaatcg tcccggtaaa 2221 tgttgtcact cgaaagaaaa ccttcgtctt cagtcaagac gagttcccga aagcgaactc 2281 aacggctgaa gcgttaggtg cattgcgccc ggccttcgat aaagcaggaa cagtcaccgc 2341 tgggaacgcg tctggtatta acgacggtgc tgccgctctg gtgattatgg aagaatctgc 2401 ggcgctggca gcaggcctta cccccctggc tcgcattaaa agttatgcca gcggtggcgt 2461 gccccccgca ttgatgggta tggggccagt acctgccacg caaaaagcgt tacaactggc 2521 ggggctgcaa ctggcggata ttgatctcat tgaggctaat gaagcatttg ctgcacagtt 2581 ccttgccgtt gggaaaaacc tgggctttga ttctgagaaa gtgaatgtca acggcggggc 2641 catcgcgctc gggcatccta tcggtgccag tggtgctcgt attctggtca cactattaca 2701 tgccatgcag gcacgcgata aaacgctggg gctggcaaca ctgtgcattg gcggcggtca 2761 gggaattgcg atggtgattg aacggttgaa ttgaggatct tgaattaagg aggacagcta 2821 aatgacaata ggtatcgata aaataaactt ttacgttcca aagtactatg tagacatggc 2881 taaattagca gaagcacgcc aagtagaccc aaacaaattt ttaattggaa ttggtcaaac 2941 tgaaatggct gttagtcctg taaaccaaga catcgtttca atgggcgcta acgctgctaa 3001 ggacattata acagacgaag acaaaaagaa aattggtatg gtaattgtgg caactgaatc 3061 agcagttgat gctgctaaag cagccgctgt tcaaattcac aacttattag gtattcaacc 3121 ttttgcacgc tgctttgaaa tgaaagaagc ttgttatgct gcaacaccag caattcaatt 3181 agctaaagat tatttagcaa ctagaccgaa tgaaaaagta ttagttattg ctacagatac 3241 agcacgttat ggattgaact caggcggcga gccaacacaa ggtgctggcg cagttgcgat 3301 ggttattgca cataatccaa gcattttggc attaaatgaa gatgctgttg cttacactga 3361 agacgtttat gatttctggc gtccaactgg acataaatat ccattagttg atggtgcatt 3421 atctaaagat gcttatatcc gctcattcca acaaagctgg aatgaatacg caaaacgtca 3481 aggtaagtcg ctagctgact tcgcatctct atgcttccat gttccattta caaaaatggg 3541 taaaaaggca ttagagtcaa tcattgataa cgctgatgaa acaactcaag agcgtttacg 3601 ttcaggatat gaagatgctg tagattataa ccgttatgtc ggtaatattt atactggatc 3661 attatattta agcctaatat cattacttga aaatcgagat ttacaagctg gtgaaacaat 3721 cggtttattc agttatggct caggttcagt tggtgaattt tatagtgcga cattagttga 3781 aggctacaaa gatcatttag atcaagctgc acataaagca ttattaaata accgtactga 3841 agtatctgtt gatgcatatg aaacattctt caaacgtttt gatgacgttg aatttgacga 3901 agaacaagat gctgttcatg aagatcgtca tattttctac ttatcaaata ttgaaaataa 3961 cgttcgcgaa tatcacagac cagagtaatt aggatctatt caggaaacag accatgtcca 4021 tgcaaagttt agataagaat tttcgacatt tatctcgtaa agaaaagtta caacaattgg 4081 ttgataagca atggttatca gaagaacaat tcgacatttt actgaatcat ccattaatcg 4141 atgaagaagt agccaatagt ttaattgaaa atgtcatcgc gcaaggtgca ttacccgttg 4201 gattattacc gaatatcatt gtggacgata aggcatatgt tgtacctatg atggtggaag 4261 agccttcagt tgtcgctgca gctagttatg gtgcaaagct agtgaatcag actggcggat 4321 ttaaaacggt atcttctgaa cgtattatga taggtcaaat cgtctttgat ggcgttgacg 4381 atactgaaaa attatcagca gacattaaag ctttagaaaa gcaaattcat aaaattgcgg 4441 atgaggcata tccttctatt aaagcgcgtg gtggtggtta ccaacgtata gcgattgata 4501 catttcctga gcaacagtta ctatctttaa aagtatttgt tgatacgaaa gatgctatgg 4561 gcgctaatat gcttaatacg attttagagg ccataactgc atttttaaaa aatgaatttc 4621 cgcaaagcga cattttaatg agtattttat ccaatcatgc aacagcgtcc gttgttaaag 4681 ttcaaggcga aattgatgtt aaagatttag caaggggcga gagaactgga gaagaggttg 4741 ccaaacgaat ggaacgtgct tctgtattgg cccaagtaga tattcatcgt gcagcaacac 4801 ataataaagg tgttatgaat ggcatacatg ctgttgtttt agcaacagga aatgatacgc 4861 gtggtgcaga agcaagtgcg catgcatacg cgagtcgtga cggacagtat cgtggtattg 4921 ctacatggcg ttacgatcaa gatcgtcaac gattgattgg tacaattgaa gtgcctatga 4981 cattggcaat cgttggcggt ggtacaaaag tattaccaat tgctaaagct tcattagagc 5041 tactaaatgt agagtcagca caagaattag gtcatgtagt tgctgccgtt ggtttagcgc 5101 aaaactttgc agcatgtcgc gcgcttgtgt cagaaggtat tcaacaaggt catatgagtt 5161 tacaatataa atcattagct atcgttgtag gggcaaaagg tgatgaaatt gctaaagtag 5221 ctgaagcttt gaaaaaagaa ccccgtgcaa atacacaagc agcggaacat attttacaag 5281 aaattagaca acaataagga tctttttaag gatctccagg catcaaataa aacgaaaggc 5341 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta 5401 gagtcacact ggctcacctt cgggtgggcc tttctgcgtt tatagcgaat tgatctggtt 5461 tgacagctta tcatcgactg cacggtgcac caatgcttct ggcgtcaggc agccatcgga 5521 agctgtggta tggctgtgca ggtcgtaaat cactgcataa ttcgtgtcgc tcaaggcgca 5581 ctcccgttct ggataatgtt ttttgcgccg acatcataac ggttctggca aatattctga 5641 aatgagctgt tgacaattaa tcatccggct cgtataatgt gtggaattgt gagcggataa 5701 caatttcagg atctaggagg aaataaccat gtctctgcca ttcctgacgt ctgcgccagg 5761 taaggtgatc atcttcggcg agcactctgc ggtgtacaat aagccggccg tcgccgcctc 5821 tgtgtctgcg ttacgcacct acctgctgat cagcgaatct tctgcaccgg acacgatcga 5881 gctggacttt ccggacatca gcttcaacca caagtggagc atcaacgact tcaacgcgat 5941 cacggaggac caggtgaaca gccaaaagct ggccaaagcc cagcaagcaa ccgacggtct 6001 gtctcaggag ctggtgtctc tgctggaccc gctgttagcg cagttaagcg agagcttcca 6061 ttaccacgcc gcgttctgct tcctgtacat gttcgtttgc ctgtgcccgc acgcaaagaa 6121 catcaagttc agcctgaaga gcacgctgcc gattggcgca ggcttaggct ctagcgcatc 6181 tatcagcgtg agcctggcgc tggcgatggc ctatctgggt ggcctgattg gcagcaacga 6241 cctggagaaa ctgagcgaaa acgacaagca catcgtgaac cagtgggcct ttatcggcga 6301 gaagtgcatt catggcaccc cgagcggcat tgacaacgca gttgccacgt atggcaacgc 6361 cctgctgttc gagaaagaca gccacaacgg cacgatcaac acgaacaact tcaagttcct 6421 ggacgacttc ccggcgatcc cgatgattct gacctacacc cgtatcccac gcagcaccaa 6481 ggatttagtc gcccgcgtgc gtgttttagt caccgaaaag ttcccggagg tgatgaagcc 6541 gatcctggac gcgatgggcg agtgcgcgct gcagggtctg gagatcatga ccaagctgag 6601 caagtgcaag ggcaccgacg atgaggcggt ggagaccaac aatgagctgt acgagcagct 6661 gctggagctg atccgtatca atcacggcct gctggtctct atcggtgtgt ctcacccggg 6721 cctggaactg atcaaaaacc tgagcgacga cctgcgcatt ggctctacga aattaacggg 6781 tgcaggtggc ggtggctgct ctttaacgct gctgcgccgt gacattacgc aggagcaaat 6841 cgacagcttc aagaagaagc tgcaggacga cttcagctac gagacgttcg agacggacct 6901 gggcggcacg ggctgttgcc tgctgagcgc caaaaatctg aacaaggacc tgaagatcaa 6961 aagcctggtg ttccagctgt tcgaaaacaa gacgaccacg aagcagcaga tcgacgacct 7021 gttactgccg ggtaacacca atctgccgtg gacgtcttaa ggatctagga gggagatcat 7081 atgagcgaat tacgtgcatt cagcgcgcca ggtaaggcac tgctggccgg tggctacctg 7141 gtgttagaca ccaagtacga ggcgttcgtc gtcggcttat ctgcccgtat gcatgcagtt 7201 gcccacccgt atggtagcct gcagggctct gacaagttcg aagtgcgtgt gaagagcaag 7261 cagttcaagg acggcgagtg gctgtaccac attagcccaa agagcggctt catcccggtt 7321 agcattggtg gcagcaagaa cccatttatc gagaaggtca ttgccaacgt cttcagctac 7381 ttcaagccga atatggacga ttactgcaac cgcaacctgt tcgtcatcga cattttcagc 7441 gacgacgcgt accacagcca agaggactct gttacggagc atcgtggtaa ccgccgcctg 7501 agcttccaca gccatcgcat tgaggaggtg ccgaagacgg gtctgggttc tagcgccggt 7561 ttagttaccg tcttaacgac ggcgttagcg agcttcttcg tgagcgacct ggagaacaac 7621 gtggacaagt accgcgaagt gattcataac ctggcgcagg tggcacattg tcaggcccaa 7681 ggtaagattg gctctggttt tgatgtggca gcggccgcct atggctctat ccgctatcgc 7741 cgctttccgc cggccctgat cagcaatctg ccggacatcg gctctgcgac gtatggtagc 7801 aaactggcgc atctggtgga cgaagaagac tggaacatca ccattaagtc taatcacctg 7861 ccgagcggct taacgttatg gatgggcgat atcaagaacg gcagcgaaac ggttaagctg 7921 gtgcagaaag tgaaaaactg gtacgacagc cacatgccgg aaagcctgaa gatttacacg 7981 gagctggacc acgccaatag ccgtttcatg gatggtctga gcaagctgga ccgcctgcac 8041 gaaacccacg acgactacag cgaccaaatc ttcgagagcc tggagcgcaa tgactgcacc 8101 tgccagaagt acccggagat cacggaggtc cgcgatgccg tggcaacgat tcgccgtagc 8161 ttccgcaaaa ttacgaagga gagcggcgcg gatatcgaac caccggtcca gacgtctctg 8221 ctggacgact gtcaaacctt aaagggcgtg ttaacgtgcc tgattccggg cgcgggtggt 8281 tacgacgcca ttgccgtcat cacgaaacag gacgtcgatc tgcgcgcaca aacggccaac 8341 gacaaacgtt tcagcaaagt ccaatggctg gatgttacgc aggccgactg gggtgttcgc 8401 aaggagaagg acccggaaac gtatctggat aagtgaggat ctaggaggat tatgagatga 8461 ccgtttacac agcatccgtt accgcacccg tcaacatcgc aacccttaag tattggggga 8521 aaagggacac gaagttgaat ctgcccacca attcgtccat atcagtgact ttatcgcaag 8581 atgacctcag aacgttgacc tctgcggcta ctgcacctga gtttgaacgc gacactttgt 8641 ggttaaatgg agaaccacac agcatcgaca atgaaagaac tcaaaattgt ctgcgcgacc 8701 tacgccaatt aagaaaggaa atggaatcga aggacgcctc attgcccaca ttatctcaat 8761 ggaaactcca cattgtctcc gaaaataact ttcctacagc agctggttta gcttcctccg 8821 ctgctggctt tgctgcattg gtctctgcaa ttgctaagtt ataccaatta ccacagtcaa 8881 cttcagaaat atctagaata gcaagaaagg ggtctggttc agcttgtaga tcgttgtttg 8941 gcggatacgt ggcctgggaa atgggaaaag ctgaagatgg tcatgattcc atggcagtac 9001 aaatcgcaga cagctctgac tggcctcaga tgaaagcttg tgtcctagtt gtcagcgata 9061 ttaaaaagga tgtgagttcc actcagggta tgcaattgac cgtggcaacc tccgaactat 9121 ttaaagaaag aattgaacat gtcgtaccaa agagatttga agtcatgcgt aaagccattg 9181 ttgaaaaaga tttcgccacc tttgcaaagg aaacaatgat ggattccaac tctttccatg 9241 ccacatgttt ggactctttc cctccaatat tctacatgaa tgacacttcc aagcgtatca 9301 tcagttggtg ccacaccatt aatcagtttt acggagaaac aatcgttgca tacacgtttg 9361 atgcaggtcc aaatgctgtg ttgtactact tagctgaaaa tgagtcgaaa ctctttgcat 9421 ttatctataa attgtttggc tctgttcctg gatgggacaa gaaatttact actgagcagc 9481 ttgaggcttt caaccatcaa tttgaatcat ctaactttac tgcacgtgaa ttggatcttg 9541 agttgcaaaa ggatgttgcc agagtgattt taactcaagt cggttcaggc ccacaagaaa 9601 caaacgaatc tttgattgac gcaaagactg gtctaccaaa ggaataagga tctaggaggt 9661 aatgataatg caaacggaac acgtcatttt attgaatgca cagggagttc ccacgggtac 9721 gctggaaaag tatgccgcac acacggcaga cacccgctta catctcgcgt tctccagttg 9781 gctgtttaat gccaaaggac aattattagt tacccgccgc gcactgagca aaaaagcatg 9841 gcctggcgtg tggactaact cggtttgtgg gcacccacaa ctgggagaaa gcaacgaaga 9901 cgcagtgatc cgccgttgcc gttatgagct tggcgtggaa attacgcctc ctgaatctat 9961 ctatcctgac tttcgctacc gcgccaccga tccgagtggc attgtggaaa atgaagtgtg 10021 tccggtattt gccgcacgca ccactagtgc gttacagatc aatgatgatg aagtgatgga 10081 ttatcaatgg tgtgatttag cagatgtatt acacggtatt gatgccacgc cgtgggcgtt 10141 cagtccgtgg atggtgatgc aggcgacaaa tcgcgaagcc agaaaacgat tatctgcatt 10201 tacccagctt aaataaggat ctcgcaaaaa accccgcttc ggcggggttt tttcgccgac 10261 tgcacggtgc accaatgctt ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg 10321 caggtcgtaa atcactgcat aattcgtgtc gctcaaggcg cactcccgtt ctggataatg 10381 ttttttgcgc cgacatcata acggttctgg caaatattct gaaatgagct gttgacaatt 10441 aatcatccgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca ggatctttta 10501 agaaggagat atacatggaa tttgacttca acaaatacat ggactccaaa gcgatgacgg 10561 taaatgaagc actgaacaaa gcgatccctc tgcgttatcc gcagaaaatc tacgaaagca 10621 tgcgttacag cctgctggca ggcggcaagc gtgttcgtcc ggttctgtgt attgccgcat 10681 gtgaactggt aggtggtacc gaagaactgg cgatcccgac cgcgtgcgca attgaaatga 10741 tccacacgat gtccctgatg cacgatgatc tgccgtgtat cgacaacgac gatctgcgtc 10801 gcggtaaacc gactaaccac aaaattttcg gtgaggatac cgcagtgact gctggtaacg 10861 cactgcactc ttacgccttc gagcatatcg cggtttctac ttctaaaacc gttggtgctg 10921 accgcatcct gcgtatggtg tccgagctgg gtcgtgctac tggctctgaa ggtgttatgg 10981 gtggtcagat ggtagacatc gcatccgaag gcgatccgtc tatcgacctg cagaccctgg 11041 aatggattca catccacaaa accgcaatgc tgctggaatg ctccgttgtt tgcggtgcaa 11101 tcattggcgg tgccagcgaa atcgtaatcg aacgtgcccg tcgctacgcc cgctgtgttg 11161 gtctgctgtt ccaggtagtt gatgacattc tggacgtaac taaaagcagc gacgaactgg 11221 gtaagactgc gggcaaggac ctgatctctg ataaagccac ctacccaaag ctgatgggtc 11281 tggaaaaggc caaggagttc tccgatgaac tgctgaaccg tgcgaagggt gaactgtcct 11341 gcttcgaccc agttaaagcc gctccgctgc tgggcctggc agactacgtg gcatttcgtc 11401 agaattaagg atcttttaag aaggagatat acatgcgtcg cagtggtaat tacaacccgt 11461 cacgttggga tgtgaatttt atccaatccc tgctgagcga ttacaaagag gacaaacacg 11521 tcattcgggc aagcgaactg gttacactgg ttaaaatgga actggaaaaa gaaacagacc 11581 aaattcgcca attagagctg attgacgact tacagcgcat ggggctgagc gatcactttc 11641 agaatgagtt caaagagatt ctgtcatcta tctatctgga ccaccattac tataaaaacc 11701 cttttccgaa agaggaacgt gatctgtata gcacaagcct ggcctttcgg ttactgcgtg 11761 agcatggctt ccaagtagca caagaggtat tcgatagctt caagaatgaa gagggcgagt 11821 tcaaagaatc actgtccgat gatactcgtg gcttattaca actgtatgaa gcgagctttc 11881 tgctgaccga gggggagacg actctggaaa gcgcacggga atttgcgacc aaattcttag 11941 aagagaaagt aaatgaaggc ggcgtagatg gcgatctgtt aacccgcatc gcgtattctc 12001 tggacattcc gttacactgg cgtattaaac ggccaaacgc gccggtgtgg attgagtggt 12061 atcgcaaacg tccagacatg aacccggtgg tgctggaatt agcaatttta gacttaaaca 12121 tcgtgcaggc tcaatttcag gaagaactga aagaatcttt tcgctggtgg cgcaacaccg 12181 gcttcgttga aaaattaccg tttgcgcgcg atcgtctggt tgaatgttat ttctggaata 12241 ccggcattat cgaaccgcgc cagcacgcga gcgcccgtat tatgatgggt aaggtcaatg 12301 cgctgattac tgtcattgac gatatttacg atgtatacgg cactctggaa gaactggagc 12361 agtttacaga tttaatccgc cggtgggaca tcaatagcat cgaccagctg ccagattaca 12421 tgcagctgtg ttttctggcg ctgaataact ttgtggacga cacctcttat gacgtcatga 12481 aagaaaaagg cgtgaatgtt atcccttatc tgcgtcagag ttgggtggat ctggccgata 12541 aatatatggt tgaggcacgc tggttctacg gtgggcacaa accgagttta gaagaatatc 12601 tggagaactc atggcagagt atcagtgggc catgcatgct gacgcatatc ttctttcgcg 12661 tcacggattc atttaccaag gagacggtcg attccttata taaataccat gacctggttc 12721 gttggtcatc tttcgtcctg cggctggcgg acgatctggg tacgtctgtg gaagaggtgt 12781 ctcgtgggga tgttccgaaa tccttacagt gctacatgtc cgattataac gcaagtgaag 12841 ctgaagctcg caaacatgtg aagtggctga tcgccgaagt gtggaagaag atgaacgctg 12901 aacgtgtttc caaggatagt ccgtttggta aggattttat tggttgcgcc gttgatctgg 12961 gtcgtatggc ccagctgatg tatcataacg gtgatggtca tgggacccag catcctatta 13021 ttcatcagca gatgacgcgt accctgtttg cacctttcgc ctaaggatcc aaactcgagt 13081 aaggatctcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt 13141 tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg 13201 gcctttctgc gtttatacct agggatatat tccgcttcct cgctcactga ctcgctacgc 13261 tcggtcgttc gactgcggcg agcggaaatg gcttacgaac ggggcggaga tttcctggaa 13321 gatgccagga agatacttaa cagggaagtg agagggccgc ggcaaagccg tttttccata 13381 ggctccgccc ccctgacaag catcacgaaa tctgacgctc aaatcagtgg tggcgaaacc 13441 cgacaggact ataaagatac caggcgtttc cccctggcgg ctccctcgtg cgctctcctg 13501 ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg gccgcgtttg tctcattcca 13561 cgcctgacac tcagttccgg gtaggcagtt cgctccaagc tggactgtat gcacgaaccc 13621 cccgttcagt ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggaa 13681 agacatgcaa aagcaccact ggcagcagcc actggtaatt gatttagagg agttagtctt 13741 gaagtcatgc gccggttaag gctaaactga aaggacaagt tttggtgact gcgctcctcc 13801 aagccagtta cctcggttca aagagttggt agctcagaga accttcgaaa aaccgccctg 13861 caaggcggtt ttttcgtttt cagagcaaga gattacgcgc agaccaaaac gatctcaaga 13921 agatcatctt attaatcaga taaaatattt ctagatttca gtgcaattta tctcttcaaa 13981 tgtagcacct gaagtcagcc ccatacgata taagttgtta ctagtgcttg gattctcacc 14041 aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga gttctgaggt 14101 cattactgga tctatcaaca ggagtccaag cgagctcgat atcaaattac gccccgccct 14161 gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac 14221 aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat 14281 atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa 14341 aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca ataaaccctt 14401 tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa 14461 actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat 14521 ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg 14581 ccatacgaaa ttccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat 14641 aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg 14701 tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc 14761 attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag 14821 ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt 14881 gaaagttgga acctcttacg tgccgatcaa cgtctcattt tcgccagata tc //