LOCUS Exported 7929 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pBbA5k-EPL122 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7929) AUTHORS Dunlop MJ, Dossani ZY, Szmidt HL, Chu HC, Lee TS, Keasling JD, Hadi MZ, Mukhopadhyay A TITLE Engineering microbial biofuel tolerance and export using efflux pumps. JOURNAL Mol Syst Biol. 2011 May 10;7:487. doi: 10.1038/msb.2011.21. PUBMED 21556065 REFERENCE 2 (bases 1 to 7929) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7929 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 4272..4289 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" terminator 4325..4396 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 4412..4439 /label=T7Te terminator /note="phage T7 early transcription terminator" primer_bind complement(4467..4484) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4601..5146) /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(4681..4700) /label=pBR322ori-F /note="pBR322 origin, forward primer" terminator 5260..5354 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(5385..6179) /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(5496..5515) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 6106..6125 /label=Neo-R /note="Neomycin resistance gene, reverse primer" CDS complement(6364..7446) /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 7408..7427 /label=LacI-R /note="LacI, reverse primer" promoter complement(7447..7524) /gene="lacI" /label=lacI promoter primer_bind 7730..7749 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" promoter 7835..7865 /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" protein_bind 7873..7889 /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)." ORIGIN 1 atgacgaaac atgccaggtt tttcctcctg ccctccttta ttctgatctc cgcggcttta 61 atcgccggtt gtaacgataa gggagaagag aaagctcacg tcggtgaacc gcaggttacc 121 gttcatattg taaaaacggc cccgttagaa gttaagactg aattaccagg ccgcaccaat 181 gcttatcgta tagccgaagt tcgcccacag gttagcggga tcgtactgaa tcgcaatttc 241 actgaaggca gcgatgtgca agcaggccag tccctgtacc agatcgatcc cgcgacctat 301 caggcaaatt atgacagcgc gaaaggcgaa ctggcgaaaa gtgaagccgc cgccgccatc 361 gcgcatttga cggtaaaacg ttacgttccg ctcgtgggta cgaaatacat cagccagcag 421 gagtacgacc aggccattgc tgatgctcgt caggccgatg ccgccgtgat tgccgcaaaa 481 gccacagtcg aaagcgctcg catcaatctt gcttatacca aagtcactgc gccaattagc 541 ggacgtatcg gcaaatcgac tgtgaccgaa ggcgctcttg tcactaatgg gcaaacgact 601 gaactggcga ctgtccagca gctcgatcct atctacgttg atgtgaccca atccagcaac 661 gattttatga ggctgaagca atccgtagag caaggaaatt tgcataagga aaacgccacc 721 agcaacgtag agttggtcat ggaaaacggt caaacctatc ccctgaaagg tacgctgcaa 781 ttctccgatg tgaccgttga tgaaagcacc ggctccataa ccctacgtgc tgtcttccct 841 aacccgcaac atacgctttt gccgggtatg tttgtgcgtg cacggattga tgaaggcgtc 901 caacctgacg ccattcttat cccgcaacaa ggcgttagcc gcacaccgcg tggtgatgca 961 accgtgctga ttgttaacga taaaagtcag gttgaagcgc gccctgtcgt tgccagtcag 1021 gcgattggcg ataaatggtt gattagtgaa ggactgaaat ctggcgatca agtcattgtc 1081 agcggcctgc aaaaagcgcg tccgggagag caggttaaag ccactaccga tacccccgca 1141 gatactgcat cgaagtaagg taatctgaca tggcaaactt ttttattcga cgaccgatat 1201 ttgcatgggt gctggccatt attctgatga tggcgggcgc actggcgatc ctacaattgc 1261 ccgtcgctca gtatccaaca attgcaccgc ctgcggtttc tgtttcagca aactatccgg 1321 gcgctgatgc gcagaccgtg caggatacgg tgacgcaggt tatcgaacag aatatgaacg 1381 gtatcgataa cctgatgtat atgtcctcca ccagcgattc cgccggtagc gtgacaatta 1441 cccttacctt ccagtccggg accgatcctg atatcgcgca agtgcaggtg cagaacaaac 1501 tccagctcgc cacgccgttg ctgccgcagg aggttcagca gcaggggatc agtgttgaaa 1561 agtccagtag cagctatttg atggtggcgg gctttgtctc tgataaccca ggcaccacac 1621 aggacgatat ctcggactat gtggcctcta acgttaaaga tacgcttagc cgtctgaatg 1681 gcgtcggtga cgtacagctt ttcggcgcac agtatgcgat gcgtatctgg ctggatgccg 1741 atctgctaaa caaatataaa ctgacaccgg ttgatgtgat taaccagttg aaggtacaga 1801 acgatcagat cgctgccgga cagttgggcg gaacgccagc gttaccaggg caacaattga 1861 acgcctcgat tattgctcag acgcggttta aaaatccgga agaattcggc aaagtgaccc 1921 tgcgcgtaaa cagtgacggc tcggtggtac gcctgaaaga tgtcgcacgg gttgaacttg 1981 gcggtgaaaa ctataacgtt atcgctcgta tcaacggaaa accggcggcg ggcctgggga 2041 ttaagctggc aaccggcgcg aatgctctcg ataccgcgaa agccattaag gcaaaactgg 2101 cggaattaca gccattcttc ccgcagggaa tgaaggttct ctacccttat gacaccacgc 2161 cattcgtcca gctttctatt cacgaagtgg taaaaacgct gttcgaagcc attatgctgg 2221 tgttcctggt gatgtatctg ttcttgcaga atatgcgagc aacgctgatc cccaccattg 2281 cggtacccgt ggtgttgtta gggacgtttg ccatcctcgc cgcttttggt tactccatca 2341 acacactaac gatgttcggg atggtgcttg ccatcgggct gctcgtcgat gatgcgatag 2401 tggtggtgga gaacgtcgag cgcgtgatga tggaggataa gctcccgcca aaagaagcga 2461 cggaaaaatc gatgtcgcaa attcagggcg cactggtggg tatcgcgatg gtgctgtcag 2521 cggtatttat tccgatggca ttcttcggcg gttctactgg ggcaatttat cgccagttct 2581 ctatcaccat cgtttcggca atggcgcttt ctgttctggt ggcattgatt cttacccctg 2641 cgttatgtgc aacgctgctt aaacccgtct ctgctgagca tcacgaaaat aagggcggtt 2701 tcttcggttg gtttaatacc accttcgatc atagcgttaa ccactacacc aacagcgtcg 2761 gcaaaatcct cggatccaca ggacgatatt tactgatcta tgcgctgatt gttgcaggaa 2821 tggtggtgtt gtttttacgt cttccgtctt ccttcttacc tgaagaggat cagggtgtct 2881 ttctgaccat gattcagtta cccgctggcg cgacgcaaga gcggacgcaa aaagtgttgg 2941 atcaagttac ggattactat ctgaagaacg agaaagcgaa cgttgaaagt gtctttacgg 3001 ttaacggctt tagcttcagc ggccaggcac aaaacgccgg tatggccttc gtcagtctga 3061 aaccgtggga agagcgtaat ggtgacgaaa acagtgcgga agcggtaatc catcgtgcca 3121 aaatggaatt gggcaagatc cgcgacggtt ttgtcattcc attcaatatg ccagccattg 3181 ttgaactggg cacggcaacg ggtttcgact ttgagttaat tgatcaggct gggctgggtc 3241 acgatgccct aacccaggcc cgtaaccagt tgcttggtat ggcggcgcaa catcctgcca 3301 gcttagtcag cgtgcgccct aatggcctgg aagacaccgc gcagtttaaa ctggaagttg 3361 accaggaaaa ggcgcaggca ttaggtgttt cactttctga catcaatcag accatttcaa 3421 cggcgctggg tgggacttac gttaacgact tcatcgaccg tggccgcgtg aaaaagttgt 3481 atgttcaggc ggatgccaaa ttccgtatgc tgccagaaga tgtcgataaa ctttatgtcc 3541 gcagcgccaa cggcgaaatg gtgccattct cggcctttac cacttcacat tgggtgtatg 3601 gctctccgcg actggaacgc tacaacggtc tgccgtcaat ggagattcag ggggaagccg 3661 cgccaggaac cagttccggc gatgccatgg cgttgatgga aaaccttgcg tcaaaattac 3721 ctgcgggcat tggttatgac tggacgggta tgtcgtatca ggaacgctta tcgggaaacc 3781 aggctcccgc tctggtagca atttcctttg tggttgtttt cctgtgcctt gctgcactct 3841 atgaaagctg gtcaattcct gtctcggtta tgttggtagt gccgttaggg attgtcggcg 3901 tgctgctggc ggcgacactc tttaatcaaa aaaatgacgt ctactttatg gtgggcttgc 3961 taacgacaat tggcttgtcg gccaaaaacg ctattttgat cgttgagttc gctaaagatc 4021 tcatggagaa agagggtaaa ggtgttgttg aagcgacact gatggcagta cgtatgcgtc 4081 tgcgtcctat cctgatgacc tctctcgcct ttattctcgg cgtattaccg ctagctatca 4141 gtaacggtgc cggcagtggc gcgcagaacg ctgtgggtat cggggtaatg ggaggaatgg 4201 tctctgcaac gttgctggca atcttcttcg taccggtgtt ctttgtggtg atccgccgtt 4261 gctttaaagg acatcatcac catcaccact gaggatccaa actcgagtaa ggatctccag 4321 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt 4381 gtcggtgaac gctctctact agagtcacac tggctcacct tcgggtgggc ctttctgcgt 4441 ttatacctag ggatatattc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga 4501 ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt tcctggaaga tgccaggaag 4561 atacttaaca gggaagtgag agggccgcgg caaagccgtt tttccatagg ctccgccccc 4621 ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg acaggactat 4681 aaagatacca ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc 4741 ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc tcattccacg cctgacactc 4801 agttccgggt aggcagttcg ctccaagctg gactgtatgc acgaaccccc cgttcagtcc 4861 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggaaag acatgcaaaa 4921 gcaccactgg cagcagccac tggtaattga tttagaggag ttagtcttga agtcatgcgc 4981 cggttaaggc taaactgaaa ggacaagttt tggtgactgc gctcctccaa gccagttacc 5041 tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt 5101 ttcgttttca gagcaagaga ttacgcgcag accaaaacga tctcaagaag atcatcttat 5161 taatcagata aaatatttct agatttcagt gcaatttatc tcttcaaatg tagcacctga 5221 agtcagcccc atacgatata agttgttact agtgcttgga ttctcaccaa taaaaaacgc 5281 ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc 5341 tatcaacagg agtccaagcg agctctcgaa ccccagagtc ccgctcagaa gaactcgtca 5401 agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg 5461 aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg 5521 tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca 5581 ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg 5641 tcgggcatgc gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct 5701 tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg 5761 cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc 5821 attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc 5881 tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc 5941 acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc 6001 agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct 6061 gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg 6121 aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg 6181 cgaaacgatc ctcatcctgt ctcttgatca gatcatgatc ccctgcgcca tcagatcctt 6241 ggcggcaaga aagccatcca gtttactttg cagggcttcc caaccttacc agagggcgcc 6301 ccagctggca attccgacgt cggtgcctaa tgagtgagct aacttacatt aattgcgttg 6361 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc 6421 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag 6481 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg 6541 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat 6601 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg caccaacgcg 6661 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag 6721 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat 6781 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt 6841 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc 6901 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg 6961 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac 7021 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat 7081 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc 7141 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat 7201 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag 7261 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc 7321 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac 7381 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg 7441 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa 7501 ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca 7561 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg 7621 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc 7681 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg 7741 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg 7801 cgtagaggat cgagatcgtt taggcacccc aggctttaca ctttatgctt ccggctcgta 7861 taatgtgtgg aattgtgagc ggataacaat ttcagaattc aaaagatctt ttaagaagga 7921 gatatacat //