质粒名称: | MP6.6TAG_AraC_Para-dnaQ926-dam-seqA-emrR-ugi-CDA1.6TAG |
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出品公司: | Addgene |
目录编号: | 163719 |
存储实验室: | Matthew Bennett |
载体骨架基本信息 |
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载体名称: | CloDF13 |
载体出品公司: | -- |
空载体大小: | -- |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | 细菌表达 |
筛选标记: | -- |
载体抗性: | 氯霉素 |
生长温度: | 37 ℃ |
宿主菌: | NEB Stable |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称: | araC dnaQ926.1TAG dam.1TAG seqA.1TAG emrR.1TAG ugi.1TAG CDA1.1TAG |
物种来源: | Synthetic |
插入大小: | -- |
基因突变: | dnaQ926.1TAG大坝。1标签序列。1标记emrR。1塔格ugi。1TAG CDA1.1TAG;每个基因的起始蛋氨酸密码子后插入琥珀色密码子 |
GenBank编号 | -- |
启动子: | -- |
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 | -- |
3’测序引物 | -- |
文献引用方法 |
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材料和方法部分: | MP6.6TAG_AraC_Para-dnaQ926-dam-seqA-emrR-ugi-CDA1.6TAG was a gift from Matthew Bennett (Addgene plasmid # 163719 ; http://n2t.net/addgene:163719 ; RRID:Addgene_163719) |
参考文献部分: | A suppressor tRNA-mediated feedforward loop eliminates leaky gene expression in bacteria. Ho JML, Miller CA, Parks SE, Mattia JR, Bennett MR. Nucleic Acids Res. 2021 Mar 18;49(5):e25. doi: 10.1093/nar/gkaa1179. 10.1093/nar/gkaa1179 PubMed 33290521 |
MP6.6TAG_AraC_Para-dnaQ926-dam-seqA-emrR-ugi-CDA1.6TAG
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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 MP6.6TAG_AraC_Para-dnaQ926-dam-seqA-emrR-ugi-CDA1.6TAG 6699 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Arabinose-inducible expression of six mutagenesis genes, each encoding one amber codon after the initiator methionine. ACCESSION . VERSION . KEYWORDS MP6.6TAG_AraC_Para-dnaQ926-dam-seqA-emrR-ugi-CDA1.6TAG SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6699) TITLE A suppressor tRNA-mediated feedforward loop eliminates leaky gene expression in bacteria. REFERENCE 2 (bases 1 to 6699) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6699 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin complement(39..777) /direction=LEFT /label=CloDF13 ori /note="Plasmids containing the CloDF13 (CDF) origin of replication can be propagated in E. coli cells that contain additional plasmids with compatible origins." terminator 867..912 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" CDS complement(927..1805) /codon_start=1 /gene="araC" /product="L-arabinose regulatory protein" /label=araC /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE EKVNDVAVKLS" promoter 1832..2116 /gene="araBAD" /label=araBAD promoter /note="promoter of the L-arabinose operon of E. coli; the araC regulatory gene is transcribed in the opposite direction (Guzman et al., 1995)" primer_bind 2039..2058 /label=pBAD Forward /note="For vectors with E. coli araBAD promoter, forward primer" CDS 4936..5193 /codon_start=1 /product="uracil-DNA glycosylase inhibitor from a Bacillus subtilis bacteriophage (Mol et al., 1995)" /label=UGI /translation="M*TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVH TAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML" CDS 5223..5852 /codon_start=1 /product="cytidine deaminase from the sea lamprey Petromyzon marinus (Rogozin et al., 2007)" /label=PmCDA1 /translation="M*TDAEYVRIHEKLDIYTFKKQFFNNKKSVSHRCYVLFELKRRGE RRACFWGYAVNKPQSGTERGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEK ILEWYNQELRGNGHTLKIWACKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKI FIQSSHNQLNENRWLEKTLKRAEKRRSELSIMIQVKILHTTKSPAV" terminator 5879..5910 /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" CDS complement(5926..6585) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVTQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" ORIGIN 1 cactcggtcg ctacgctccg ggcgtgagac tgcggcgggc gctgcggaca catacaaagt 61 tacccacaga ttccgtggat aagcagggga ctaacatgtg aggcaaaaca gcagggccgc 121 gccggtggcg tttttccata ggctccgccc tcctgccaga gttcacataa acagacgctt 181 ttccggtgca tctgtgggag ccgtgaggct caaccatgaa tctgacagta cgggcgaaac 241 ccgacaggac ttaaagatcc ccaccgtttc cggcgggtcg ctccctcttg cgctctcctg 301 ttccgaccct gccgtttacc ggatacctgt tccgcctttc tcccttacgg gaagtgtggc 361 gctttctcat agctcacaca ctggtatctc ggctcggtgt aggtcgttcg ctccaagctg 421 ggctgtaagc aagaactccc cgttcagccc gactgctgcg ccttatccgg taactgttca 481 cttgagtcca acccggaaaa gcacggtaaa acgccactgg cagcagccat tggtaactgg 541 gagttcgcag aggatttgtt tagctaaaca cgcggttgct cttgaagtgt gcgccaaagt 601 ccggctacac tggaaggaca gatttggttg ctgtgctctg cgaaagccag ttaccacggt 661 taagcagttc cccaactgac ttaaccttcg atcaaaccac ctccccaggt ggttttttcg 721 tttacagggc aaaagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 781 tctactgaac cgctctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc 841 agcccaggag gaagaggaca tccggtcaaa taaaacgaaa ggctcagtcg aaagactggg 901 cctttcgttt tagacttagg gaccctttat gacaacttga cggctacatc attcactttt 961 tcttcacaac cggcacggaa ctcgctcggg ctggccccgg tgcatttttt aaatacccgc 1021 gagaaataga gttgatcgtc aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg 1081 gtggtgctca aaagcagctt cgcctggctg atacgttggt cctcgcgcca gcttaagacg 1141 ctaatcccta actgctggcg gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc 1201 tgtgcgacgc tggcgatatc aaaattgctg tctgccaggt gatcgctgat gtactgacaa 1261 gcctcgcgta cccgattatc catcggtgga tggagcgact cgttaatcgc ttccatgcgc 1321 cgcagtaaca attgctcaag cagatttatc gccagcagct ccgaatagcg cccttcccct 1381 tgcccggcgt taatgatttg cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc 1441 gggcgaaaga accccgtatt ggcaaatatt gacggccagt taagccattc atgccagtag 1501 gcgcgcggac gaaagtaaac ccactggtga taccattcgc gagcctccgg atgacgaccg 1561 tagtgatgaa tctctcctgg cgggaacagc aaaatatcac ccggtcggca aacaaattct 1621 cgtccctgat ttttcaccac cccctgaccg cgaatggtga gattgagaat ataacctttc 1681 attcccagcg gtcggtcgat aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa 1741 cccgccacca gatgggcatt aaacgagtat cccggcagca ggggatcatt ttgcgcttca 1801 gccatacttt tcatactccc accattcaga gaagaaacca attgtccata ttgcatcaga 1861 cattgccgtc actgcgtctt ttactggctc ttctcgctaa cccaaccggt aaccccgctt 1921 attaaaagca ttctgtaaca aagcgggacc aaagccatga caaaaacgcg taacaaaagt 1981 gtctataatc acggcagaaa agtccacatt gattatttgc acggcgtcac actttgctat 2041 gccatagcat ttttatccat aagattagcg gatcctacct gacgcttttt atcgcaactc 2101 tctactgttt ctccataccc gtttttttgg acgcgtacaa ctcaagtctg acataaatga 2161 ccgctatgta gagcactgca attacacgcc agatcgttct cgctaccgca accaccggta 2221 tgaaccagat tggtgcgcac tatgaaggcc acaagatcat tgagattggt gccgttgaag 2281 tggtgaaccg tcgcctgacg ggcaataact tccatgttta tctcaaaccc gatcggctgg 2341 tggatccgga agcctttggc gtacatggta ttgccgatga atttttgctc gataagccca 2401 cgtttgccga agtagccgat gagttcatgg actatattcg cggcgcggag ttggtgatcc 2461 ataacgcagc gttcgatatc ggctttatgg actacgagtt ttcgttgctt aagcgcgata 2521 ttccgaagac caatactttc tgtaaggtca ccgatagcct tgcggtggcg aggaaaatgt 2581 ttcccggtaa gcgcaacagc ctcgatgcgt tatgtgctcg ctacgaaata gataacagta 2641 aacgaacgct gcacggggca ttactcgatg cccagatcct tgcggaagtt tatctggcga 2701 tgaccggtgg tcaaacgtcg atggcttttg cgatggaagg agagacacaa cagcaacaag 2761 gtgaagcaac aattcagcgc attgtacgtc aggcaagtaa gttacgcgtt gtttttgcga 2821 cagatgaaga gattgcagct catgaagccc gtctcgatct ggtgcagaag aaaggcggaa 2881 gttgcctctg gcgagcataa tttaatatca gtaaaccgga cataacccat gtagaagaaa 2941 aatcgcgctt ttttgaagtg ggcagggggc aagtatcccc tgcttgatga tattaaacgg 3001 catttgccca agggcgaatg tctggttgag ccttttgtag gtgccgggtc ggtgtttctc 3061 aacaccgact tttctcgtta tatccttgcc gatatcaata gcgacctgat cagtctctat 3121 aacattgtga agatgcgtac tgatgagtac gtacaggccg cacgcgagct gtttgttccc 3181 gaaacaaatt gcgccgaggt ttactatcag ttccgcgaag agttcaacaa aagccaggat 3241 ccgttccgtc gggcggtact gtttttatat ttgaaccgct acggttacaa cggcctgtgt 3301 cgttacaatc tgcgcggtga gtttaacgtg ccgttcggcc gctacaaaaa accctatttc 3361 ccggaagcag agttgtatca cttcgctgaa aaagcgcaga atgccttttt ctattgtgag 3421 tcttacgccg atagcatggc gcgcgcagat gatgcatccg tcgtctattg cgatccgcct 3481 tatgcaccgc tgtctgcgac cgccaacttt acggcgtatc acacaaacag ttttacgctt 3541 gaacaacaag cgcatctggc ggagatcgcc gaaggtctgg ttgagcgcca tattccagtg 3601 ctgatctcca atcacgatac gatgttaacg cgtgagtggt atcagcgcgc aaaattgcat 3661 gtcgtcaaag ttcgacgcag tataagcagc aacggcggca cacgtaaaaa ggtggacgaa 3721 ctgctggctt tgtacaaacc aggagtcgtt tcacccgcga aaaaataatt cagctaagac 3781 actgcactgg attaagatgt agaaaacgat tgaagttgat gatgaactct acagctatat 3841 tgccagccac actaagcata tcggcgagag cgcatccgac attttacggc gtatgttgaa 3901 attttccgcc gcatcacagc ctgctgctcc ggtgacgaaa gaggttcgcg ttgcgtcacc 3961 tgctatcgtc gaagcgaagc cggtcaaaac gattaaagac aaggttcgcg caatgcgtga 4021 acttctgctt tcggatgaat acgcagagca aaagcgagcg gtcaatcgct ttatgctgct 4081 gttgtctaca ctatattctc ttgacgccca ggcgtttgcc gaagcaacgg aatcgttgca 4141 cggtcgtaca cgcgtttact ttgcggcaga tgaacaaacg ctgctgaaaa atggtaatca 4201 gaccaagccg aaacatgtgc caggcacgcc gtattgggtg atcaccaaca ccaacaccgg 4261 ccgtaaatgc agcatgatcg aacacatcat gcagtcgatg caattcccgg cggaattgat 4321 tgagaaggtt tgcggaacta tctaacggct gaaattaatg aggtcatacc caaatgtagg 4381 atagttcgtt tacgcccatt gaacaaatgc taaaatttcg cgccagccgc cacgaagatt 4441 ttccttatca ggagatcctt ctgactcgtc tttgcatgca catgcaaagc aagctgctgg 4501 agaaccgcaa taaaatgctg aaggctcagg gaattaacga gacgttgttt atggcgttga 4561 ttacgctgga gtctcaggaa aaccacagta ttcagccttc tgaattaagt tgtgctcttg 4621 gatcatcccg taccaacgcg acgcgtattg ccgatgaact ggaaaaacgc ggttggatcg 4681 aacgtcgtga aagcgataac gatcgccgct gcctgcatct gcaattaacg gaaaaaggtc 4741 acgagttttt gcgcgaggtt ttaccaccgc agcataactg cctgcatcaa ctctggtccg 4801 cgctcagcac aacagaaaaa gatcagctcg agcaaatcac ccgcaaattg ctctcccgtc 4861 tcgaccagat ggaacaagac ggtgtggttc tcgaagcgat gagctaataa tacaaaaatt 4921 aggaggaatt tcaacatgta gacaaattta tctgacatca ttgaaaaaga aacaggaaaa 4981 caactagtga ttcaagaatc aattctaatg ttaccagaag aagtagagga agtaattggg 5041 aataaaccag aaagtgatat tttagttcat actgcttatg atgaaagtac agatgaaaat 5101 gtaatgctat taacttcaga tgctccagaa tataaacctt gggctttagt aattcaagac 5161 agtaatggag aaaataaaat taaaatgtta taagtcgaga ttaagtaaac cggaatctga 5221 agatgtagac cgacgcggaa tacgttcgta tccacgaaaa actggacatc tacaccttca 5281 aaaaacagtt cttcaacaac aaaaaatctg tttctcaccg ttgctacgtt ctgttcgaac 5341 tgaaacgtcg tggtgaacgt cgtgcgtgct tctggggtta cgcggttaac aaaccgcagt 5401 ctggtaccga acgtggtatc cacgcggaaa tcttctctat ccgtaaagtt gaagaatacc 5461 tgcgtgacaa cccgggtcag ttcaccatca actggtactc ttcttggtct ccgtgcgcgg 5521 actgcgcgga aaaaatcctg gaatggtaca accaggaact gcgtggtaac ggtcacaccc 5581 tgaaaatctg ggcgtgcaaa ctgtactacg aaaaaaacgc gcgtaaccag atcggtctgt 5641 ggaacctgcg tgacaacggt gttggtctga acgttatggt ttctgaacac taccagtgct 5701 gccgtaaaat cttcatccag tcttctcaca accagctgaa cgaaaaccgt tggctggaaa 5761 aaaccctgaa acgtgcggaa aaacgtcgtt ctgaactgtc tatcatgatc caggttaaaa 5821 tcctgcacac caccaaatct ccggcggttt aaacttaatt aacggcactc ctcagccaag 5881 tcaaaagcct ccggtcggag gcttttgact acatgcccat ggcgtttacg ccccgccctg 5941 ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca 6001 aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata 6061 tttgcccata gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa 6121 actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt 6181 agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa 6241 ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg 6301 gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc 6361 catacggaac tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata 6421 aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt 6481 ctggttatag gtacattgag taactgactg aaatgcctca aaatgttctt tacgatgcca 6541 ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc 6601 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg 6661 aaagttggaa cctcttacgt gccaagccaa ataggccgt //