质粒名称: | pET28a_T7-ARG1 |
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出品公司: | 爱迪基因 |
目录编号: | 106473 |
存储实验室: | Mikhail Shapiro |
载体骨架基本信息 |
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载体名称: | pET28a |
载体出品公司: | Novagen |
载体抗性: | 卡那霉素 |
空载体大小: | 5374 bp |
完整质粒大小: | 12129 bp |
载体修饰: | -- |
载体类型: | 细菌表达 |
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: | ARG1,Acoustic Reporter Gene 1,Hybrid Gas Vesicle gene cluster |
物种来源: | Anabaena flos-aquae and Bacillus megaterium |
插入大小: | 6755 bp |
基因突变: | -- |
GenBank编号 | -- |
启动子 | T7 |
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 | T7_Forward |
3' 测序引物 | T7 terminator_Reverse |
文献引用方法 |
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材料和方法部分: | pET28a_T7-ARG1 was a gift from Mikhail Shapiro (Addgene plasmid # 106473 ; http://n2t.net/addgene:106473 ; RRID:Addgene_106473) |
参考文献部分: | Acoustic reporter genes for noninvasive imaging of microorganisms in mammalian hosts. Bourdeau RW, Lee-Gosselin A, Lakshmanan A, Farhadi A, Kumar SR, Nety SP, Shapiro MG. Nature. 2018 Jan 3;553(7686):86-90. doi: 10.1038/nature25021. 10.1038/nature25021 PubMed 29300010 |
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载体基本信息
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载体简介
载体序列
LOCUS pET28a_T7-ARG1 12125 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses acoustic reporter genes or gas vesicles (ARG1) in Escherichia coli. ACCESSION . VERSION . KEYWORDS pET28a_T7-ARG1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12125) AUTHORS Bourdeau RW, Lee-Gosselin A, Lakshmanan A, Farhadi A, Kumar SR, Nety SP, Shapiro MG TITLE Acoustic reporter genes for noninvasive imaging of microorganisms in mammalian hosts. JOURNAL Nature. 2018 Jan 3;553(7686):86-90. doi: 10.1038/nature25021. PUBMED 29300010 REFERENCE 2 (bases 1 to 12125) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..12125 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 12..467 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(99..118) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 309..330 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(559..1374) /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 1282..1301 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 1366..1385 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 1496..2084 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1985..2004 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2238..2255 /label=L4440 /note="L4440 vector, forward primer" misc_feature 2270..2409 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(2335..2354) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2495..2517 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(2511..2702) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(3511..4593) /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 4555..4574 /label=LacI-R /note="LacI, reverse primer" promoter complement(4594..4671) /gene="lacI" /label=lacI promoter primer_bind 4877..4896 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind 4980..4999 /label=T7 /note="T7 promoter, forward primer" promoter 4980..4998 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 4999..5023 /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 8305..8322 /codon_start=1 /product="N-myristoylation signal from Src kinase (Pellman et al., 1985; Kaplan et al., 1988)" /label=myr /translation="GSSKSK" primer_bind complement(12039..12057) /label=T7 Term /note="T7 terminator, reverse primer" terminator 12053..12100 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" ORIGIN 1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 421 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt 481 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat 541 ccgctcatga attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt 601 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa 661 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg 721 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa 781 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca 841 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc 901 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca 961 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt 1021 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt 1081 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat 1141 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc 1201 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt 1261 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat 1321 gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcataacacc 1381 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgaccaaa atcccttaac 1441 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag 1501 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg 1561 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca 1621 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga 1681 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca 1741 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc 1801 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca 1861 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa 1921 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc 1981 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc 2041 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg 2101 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat 2161 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca 2221 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt 2281 attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat 2341 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc 2401 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc 2461 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt 2521 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga 2581 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc 2641 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc 2701 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga 2761 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt 2821 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa 2881 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg 2941 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa 3001 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag 3061 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc 3121 cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ggccgccatg 3181 ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct 3241 tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc 3301 cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt 3361 tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg 3421 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga 3481 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 3541 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 3601 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc 3661 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc 3721 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc 3781 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc 3841 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat 3901 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg 3961 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac 4021 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc 4081 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc 4141 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc 4201 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg 4261 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc 4321 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc 4381 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac 4441 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc 4501 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata 4561 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc 4621 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc 4681 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt 4741 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg 4801 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag 4861 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc 4921 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt 4981 aatacgactc actatagggg aattgtgagc ggataacaat tccaagcttc gccagggttt 5041 tcccagtcga gctcatgact catatttata gttattctcc tcatttccca agtttccgac 5101 aacaaataac aaggaattaa aaaaatggca gttgaaaaaa ccaattcttc ctccagcttg 5161 gcagaagtta ttgatagaat cctcgacaaa ggtatcgtaa ttgacgcttg ggttcgtgtt 5221 tccttagttg gtatcgaact actagcaatt gaagctcgga tcgttatcgc ttccgttgaa 5281 acctacttga agtatgctga agcagttggt ttgacccaat cagcagcagt acctgcttaa 5341 tttaattaag caagttacca agtaggagaa taaccatgac tcatatttat agttattctc 5401 ctcatttccc aagtttccga caacaaataa caaggaatta aaaaaatggc agttgaaaaa 5461 accaattctt cctccagctt ggcagaagtt attgatagaa tcctcgacaa aggtatcgta 5521 attgacgctt gggttcgtgt ttccttagtt ggtatcgaac tactagcaat tgaagctcgg 5581 atcgttatcg cttccgttga aacctacttg aagtatgctg aagcagttgg tttgacccaa 5641 tcagcagcag tacctgctta atttaattaa gctccataat atcaggatta attaatcctg 5701 atattggttc tgcttttttt aattatctag atatttggaa atcaacgctt ctcaaaggat 5761 attacaaaaa agtacgaatc cttataatgt agtacctgga gaagcaattt ctgttaggtc 5821 tgggagctcc catgggatcg ttatcccgcg gccaagcttg ctagcgctaa tttactattt 5881 tttggagaac ttcatgattt ctttaatggc aaaaatccgg caagaacatc agtcaatagc 5941 agagaaagtg gctgaactat ctcttgagac cagagaattc ttgtccgtca cgacagcgaa 6001 aagacaagag caagctgaaa aacaagctca agaactgcaa gcattctaca aggatcttca 6061 ggaaacaagt cagcagtttt tatcagaaac agcccaagcc agaattgctc aagctgaaaa 6121 acaagctcaa gaactgttag cattccacaa agaacttcaa gaaacaagtc agcagttttt 6181 atcagcaaca gcccaagcca gaattgctca agctgaaaaa caagcgcaag aactgttagc 6241 attttatcaa gaagttcggg aaacaagtca gcagttttta tcagcaacag cccaagcaag 6301 aattgctcaa gctgaaaaac aagctcaaga actgttagca ttccacaaag aacttcaaga 6361 aacaagtcag cagtttttat cagcaacagc cgacgcaaga actgctcaag ctaaggaaca 6421 gaaggaatct ctcctgaaat tccgtcagga tttgtttgtg agtatctttg gttaataaag 6481 atctaactat tggaggctac taaaaatgga aattaaaaaa attatgcaag ccgtgaacga 6541 ctttttcggt gaacacgtag ctcctcctca taaaattacc tcggtggaag ctactgaaga 6601 tgaaggttgg agagttattg ttgaagtcat tgaagaacga gaatatatga aaaaatacgc 6661 caaagatgaa atgctcggaa cgtacgagtg ctttgtaaat aaagaaaaag aagtcatttc 6721 attcaaacga ctcgacgtca gatatagaag cgccattggc attgaagcat aaacgataag 6781 atggcaggag gaacgtaaaa atgaccgtct taacagacaa aaggaaaaaa ggcagtggag 6841 cttttataca agatgacgag acaaaagagg ttctttcaag agcgctgagc tatttaaaat 6901 ccggctattc cattcatttt acaggtcctg ccggcggagg caaaacctct ttagcgcgag 6961 cgcttgctaa aaagagaaag cgtcctgtaa tgctgatgca cgggaatcac gagctcaaca 7021 acaaagattt aattggcgat tttacgggat acacgagcaa aaaagtaatc gaccagtacg 7081 ttcgttctgt ctataaaaaa gatgaacagg tgagtgaaaa ctggcaggat ggccgattgc 7141 ttgaagctgt aaaaaatggc tatacgctga tttacgacga atttactcgt tctaagcctg 7201 cgacgaataa tatctttcta tcgatattag aagaaggcgt gctgccgctg tatggagtaa 7261 aaatgaccga tccttttgtg cgcgtgcatc ccgatttccg cgtcatcttc acaagcaatc 7321 cagctgagta tgccggcgta tatgatacgc aagatgcgct tctcgacagg ttaattacca 7381 tgtttattga ttataaagac atcgacagag agacagcgat tttaacggag aaaacggacg 7441 tagaagaaga tgaagcgcgc acaattgtaa cgctcgtagc aaacgtgcga aaccgctctg 7501 gagacgaaaa cagcagcgga cttagcctgc gggcttcgct tatgatcgct acccttgcca 7561 cgcagcaaga cattcctatc gatggaagtg acgaagattt tcaaacgtta tgtatcgata 7621 ttttgcatca tccgcttacc aaatgtttgg atgaagaaaa tgcaaaaagc aaagccgaaa 7681 aaatcatttt agaagaatgt aagaatatag acactgaaga aaagtaaagg agcttgaaaa 7741 catgagtgaa acaaacgaaa caggtattta tatttttagc gccattcaaa cggataaaga 7801 cgaagaattt ggcgccgtgg aagtagaagg aacaaaagct gaaacatttt tgattcgcta 7861 caaagacgcg gctatggtag cagctgaagt accgatgaaa atttatcatc ctaatcgcca 7921 aaatttatta atgcatcaaa acgcagtagc agcgattatg gacaagaacg atacggttat 7981 tccaatcagc tttgggaatg tattcaaatc aaaagaagac gtaaaagttc ttttggaaaa 8041 cctttatccg cagtttgaaa agctgtttcc agcgatcaaa ggaaaaattg aagtcggttt 8101 aaaagtaatt gggaaaaaag aatggcttga gaaaaaagta aacgaaaatc ctgaacttga 8161 gaaagtatca gcatccgtaa aaggaaaatc agaagcagcc ggttattatg agcgtattca 8221 acttggagga atggctcaaa agatgtttac ttccctgcaa aaagaagtca agacagatgt 8281 gttttctccg cttgaagaag cagcggaagc agcaaaagca aatgagccaa cgggcgaaac 8341 gatgctttta aacgcgtctt tcttaattaa ccgagaagat gaagcgaagt ttgatgaaaa 8401 agtaaatgaa gcgcatgaaa actggaaaga caaagccgat tttcattaca gcggtccttg 8461 gcctgcttat aattttgtga acattcgcct aaaagtagaa gagaaataac gtgcttcaca 8521 aattagtaac cgcacccatt aaccttgtag tgaaaatcgg cgaaaaagta caggaagaag 8581 ctgataaaca gctatatgac cttccgacga ttcagcaaaa gctcattcag cttcaaatga 8641 tgtttgagct tggtgaaatt ccagaagaag cgtttcaaga aaaagaagat gaattgttaa 8701 tgaggtacga aattgcgaaa cgcagagaaa ttgaacaatg ggaagagcta acacaaaaaa 8761 gaaatgagga atcctagatg ggagaattac tgtatttata cggtttaatt ccaacaaaag 8821 aagcagcagc catagagccg tttccatctt ataaggggtt tgacggagaa cattcactgt 8881 acccaattgc gtttgatcag gtgacggctg tagtttctaa gctggatgct gacacctatt 8941 cagaaaaagt gattcaagaa aaaatggagc aggatatgag ctggctgcag gaaaaagcat 9001 ttcatcatca cgaaacggta gccgctttgt acgaagaatt tacgatcatt ccattaaaat 9061 tttgcaccat ttataaaggt gaagaaagtc tgcaggcagc tattgagatt aacaaagaaa 9121 agatagagaa ttcactgacg ctgcttcaag gaaatgaaga gtggaatgtg aaaatttact 9181 gtgatgatac agagcttaaa aaaggaatca gcgaaacgaa tgaaagcgtg aaagcgaaaa 9241 aacaagaaat tagtcactta tcaccaggaa gacagttttt tgaaaagaaa aaaatagatc 9301 agctgattga aaaagaatta gagcttcaca aaaacaaagt gtgtgaagag atacatgaca 9361 agctaaaaga attatcgctt tatgactctg ttaaaaagaa ttggagcaaa gacgtaactg 9421 gcgcagctga acagatggcg tggaacagcg tgtttcttct cccgtctctg cagattacta 9481 agttcgtaaa cgaaatagaa gagcttcagc aaaggcttga aaataaaggc tggaagtttg 9541 aagtgacggg accatggccg ccctatcatt tctcgagctt tgcgtaaagt gaggaattaa 9601 cattatgtct cttaaacaat ccatggagaa taaagatatt gctcttattg atattttaga 9661 tgtcatttta gataaaggag tcgccattaa aggagactta atcatttcca tagctggcgt 9721 cgatttagtg tatttggatt tgcgggtgct tatttcttcg gttgaaacgc ttgtgcaagc 9781 aaaagaagga aatcacaaac caatcacttc tgaacaattt gataaacaaa aggaggaatt 9841 aatggatgca accggtcagc caagcaaatg gacgaatcca cttggatcct gatcaagctg 9901 aacaaggctt agcgcagctt gtgatgacag ttattgagct attgaggcaa atcgttgaac 9961 gtcatgccat gaggcgggtg gagggtggaa cgttgacgga cgaacaaatt gaaaacttag 10021 gaattgcact aatgaactta gaagaaaaaa tggacgagtt gaaagaggtg ttcggtctgg 10081 atgcagaaga tttaaatatt gatcttggac cgctaggcag cctgctttaa gcggtcagta 10141 ggaggaacag tatggcagtc gaacataata tgcagtcaag tacgattgta gatgtgctcg 10201 aaaagatttt ggataaagga gtcgttatag cgggggacat caccgtagga attgcagatg 10261 tcgagctatt aacgataaag atccgcttga ttgtggcttc ggttgataag gcaaaagaaa 10321 tcggcatgga ctggtgggaa aatgatccgt atctcagttc aaaaggagcc aataacaaag 10381 cgctcgaaga agaaaataaa atgctgcatg agcggttaaa aacgcttgaa gaaaaaatag 10441 aaacgaaacg ttaaaaactg tacgctactt aaaaaatgga gggatttaca atggcaactg 10501 aaacaaaatt agataacaca caggcagaaa acaaggaaaa taaaaatgcg gaaaacggtt 10561 caaaagaaaa gaacggttca aaagcaagca aaacaacaag cagcgggcca atcaaacgag 10621 cggtagcagg aggcatcatc ggtgcaacga ttggatatgt atcgactcct gaaaatcgaa 10681 aaagtctcct tgaccgcatt gatacagacg aattaaaaag caaagcatct gatttaggaa 10741 caaaggtaaa agaaaaatca aaaagcagcg tggccagcct gaaaacatct gcgggaagct 10801 tgtttaaaaa agataaagat aaatcaaaag atgatgaaga aaacgtaaat tcttctagta 10861 gcgaaacaga agacgataac gttcaagagt acgacgagtt aaaagaagaa aatcaaactc 10921 ttcaagatcg cttatcacag cttgaagaaa aaatgaacat gcttgttgag cttagcctca 10981 ataaaaatca agacgaagaa gcggaagata cagattccga cgaagaagag aacgatgaga 11041 acgatgaaaa cgatgaaaac gagcaggacg atgaaaacga agaagaaaca tctaagccac 11101 gtaaaaagga taaaaaagaa gctgaggaag aagaaagtga aagtgacgaa gacagcgagg 11161 aagaagagga agattctcgc tcaaacaaaa aaaataaaaa agtaaaaaca gaagaagaag 11221 acgaagatga aagcgaagaa gaaaaaaagg aagcgaaacc aaaaaagtca acagctaaaa 11281 aatcaaaaaa tacaaaagca aagaaaaaca cggacgaaga agatgatgaa gcaacatctc 11341 tttctagtga agacgataca acagcctaag acgtaaagga ggaaagaaag acatgagtac 11401 aggcccttct ttttcaacta aagacaatac gcttgaatac tttgtgaaag cttctaataa 11461 acacggcttt tcacttgata tttcattaaa tgtaaacggc gctgtgattt ccggtaccat 11521 gatttcagca aaagaatact ttgattactt aagcgaaacg tttgaagaag gcagtgaagt 11581 ggctcaggcg ctaagcgaac aattctcttt agcaagcgaa gcgagcgaat caaacggaga 11641 agcagaagcc cattttattc atttgaaaaa tacaaagatt tactgtggag acagtaaatc 11701 tactccttct aaaggcaaaa tcttttggag agggaaaata gcagaagtag acgggttttt 11761 cttaggaaag atttctgatg caaaatcaac gagtaaaaag agttcataaa aaacggcggg 11821 gtgattgccc cgccgttttt tagtgatgtg atgagatgtg cagcttcttt tttcgataca 11881 tgagcggagt catcttcatt tcttttcgaa actgattaat aaaatagctt gtactattaa 11941 accctacttg ataggcgact tccgtcacat ttgcttctgc ttgctgcagg atccggctgc 12001 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata 12061 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc 12121 cggat //
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