LOCUS Exported 8145 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pBbA5k-EPL4 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8145) 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 8145) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8145 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 4488..4505 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" terminator 4541..4612 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 4628..4655 /label=T7Te terminator /note="phage T7 early transcription terminator" primer_bind complement(4683..4700) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4817..5362) /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(4897..4916) /label=pBR322ori-F /note="pBR322 origin, forward primer" terminator 5476..5570 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(5601..6395) /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(5712..5731) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 6322..6341 /label=Neo-R /note="Neomycin resistance gene, reverse primer" CDS complement(6580..7662) /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 7624..7643 /label=LacI-R /note="LacI, reverse primer" promoter complement(7663..7740) /gene="lacI" /label=lacI promoter primer_bind 7946..7965 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" promoter 8051..8081 /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" protein_bind 8089..8105 /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 atgggcatcc tgaacggacg gtcgtcgaac ccggcaagcg gcggagcgcg ctcccggcgc 61 ccgtggcgtt cgctcgccgc tccgctcgcc ggcctcgccc tgctcgccgg ttgcggggaa 121 accgccgccc cgccggcgcg cccacccacc gaggtgaccg ccatcacgct ggccgcgcgg 181 gataccccgg tggtcttcga attcgtcgcc cagacccaga gttcccggga agtggaaatc 241 cgcgcccggg tcgagggctt cctggaaaaa cgcctgtaca acgaaggcga tctggtgcgc 301 accggacaga tcctgttcca gatggacccc aggccgttcg aggcggcctt gcagtcggcc 361 cgcggccagc tcgcccagca gcaggcgcgc ctggatatgg ccagggccga cctgggccgc 421 atccgtcccc tggtgaaggt gggcgcgctg agcaagaaaa acctcgacga cgccatcggc 481 accgagcggc aggcgcaggc cgccgtgctc tccgccgagg gcgaggtgcg cacggcgcag 541 ctcaatctcg gctacaccac catcaagtcg ccgctcaccg gggtgtcgag cttcgccaag 601 ctgcaggaag gcagctatgt gacgccgggc gcggccggcc tgctgactta cgtgtcgcag 661 gtcgacccca tctacatcaa cttcagcctc tcggagaacg agatgctgaa attccgcggc 721 gagatcgccg ccggccagct caggtttccc ccgcacaacg atttcgaggt cgaggtggtg 781 ctggccgacg gctcgcaggt gcccgatcgc ggccgcctcg acttcgccga cttcgccttc 841 agcgaggaaa ccggcacctt cctgatccgg gcggaaatgg ccaacccgaa gggcctgctg 901 cgccccggcc agttcgtgcg cgcccacctc aagggcgcgg tccgccctgg cgccctgctg 961 gtgccccagc gcgcggtcct gcagggttcc aagagccatt acgtgtgggt gatcggcgac 1021 ggcggcaagc ccgaacagcg catcgtggaa atgggcgact ggcacggcga cgactggttc 1081 gtcgcccggg gcctgcgcgc cggcgagcgc atcgtggtcg acggtgcgat ccgggtgacg 1141 ccggaggctt cgctgcagat cgtcgacgcc cgtcccgccg ccggggaaac cggccagccg 1201 caacggcaga aggcggccga ggcctggccc ggccagcagg agcggttgaa ggccggcccg 1261 tccggcgggg gacaggcgcc gtgagcctgt cccactactg catcgagcgg ccgatcttcg 1321 cctcggtgat ttccatcgtc atcaccctgg ccggggcagt ggcgatgttc aacctgccgg 1381 tggcccagta cccggacatc accccgccgc agatcaccgt atcggccacc tatcccggcg 1441 ccgacgccga agtggtggcc aacaacgtgg cggcgcccat cgagcagcag gtcaacggcg 1501 ccgacgacat gatctacatg aactcgtcga gttcggcgac cggcaacatg accctcaacg 1561 tgttcttcga gatcggcacc gatccctcgc tggcccaggt ggacgtgcag aaccgggtca 1621 acctggccct gccgcaactg ccctcggcgg tgcagagcca gggcatccag gtgcagaaga 1681 agtcctcggc cttcatgatg gtcatcgcgg tctactcccc ggacgaacgc tacgacagca 1741 cctacatcgc caactacgcc aacctgtaca ttctcgacgc gatcaagcgc atccccggcg 1801 ccaaccaggc gagcatcttc ggcaccccgg actacgccat gcgcatctgg ctcaagcccg 1861 accgcatggc ccagttgaag atcaccgccg ccgacgtgca gcaggcggtg gccaaccaga 1921 accagcagtt cgccgtgggc cgcatcggcc agtcgccgac cggcgaggcg gtggagcagt 1981 ccttcgccgt caccaccaag ggccggctga ccgagccgga ggagttcgag aacatcatca 2041 tccgcgccgc cagcggcggg gcggccatcg tccgcctgcg cgacgtgggc cgcgccgagc 2101 tgggccagaa ggactactcc ctgcgcagca cctaccaggg caagccggcg acgctgatgg 2161 cggtgtacca gcagcccggc gccaatgccc tggacgtgtc cgacgccgtc aggacgaccc 2221 tggcggagat ggagaagagt tttcccgccg gcatcgaata caagatcgtg atggacacca 2281 cggccttcac ccgcacctcc atcgccgagg tggtgcacac cttcttcgag gccctggtgc 2341 tggtggtggc ggtcgtttac ctgttcctgc agagcctgcg cgccaccctg atcccggtgc 2401 tggcagtgcc ggtgtcgatc gtcggcacct tcatcggcat gaccgccctg ggcttctcga 2461 tcaacatgct gaccctgttc ggcatggtgc tggcgatcgg catcgtggtg gacgacgcca 2521 tcgtggtgat cgagaacgtc gagcgcaaca tgcaggtcca ccgcctgccg cccagggagg 2581 ccgccaagcg ggcgatggac gaggtggccg gcccggtggt ggccatcgtc ctggtgctct 2641 gcgcggtgtt cgtcccggtg gccttcatgg gcggcatcac cggccagctc tacaagcagt 2701 tcgccatcac catcgccatc tccgtggtga tctccggcct ggtcgccctg accctgtcgc 2761 cggccctcgc cgcgatcctc atcaagcccg cccacgggcg caagcgcggc ttcttccgct 2821 ggttcgaacg caacttcgag cgcatgaccg agggctactc gcgcgcggtg gccttcatga 2881 tcaagcgctt cctgctcgcc ctgctgctgt tcgccgggat ggtcgtgctc accgtgctga 2941 tggcgcagcg catccccggc gccttcctgc cgccggagga ccagggctac ctgctcggcg 3001 cggtgatcat gcccgacgcc gccagcctgg accgcaccgg cgaggtgggc cgggcggtca 3061 gcgactactt catgcgcgac gaagccgtcg agggggtggc catcgtcaac ggctacagcc 3121 tgctcgacag ccagaacaag aacaacgccg gcaccttctt cgtcggtttc aaggacttcg 3181 aggaacgcta caaggacgac gaaagcatcc gtacgcagag cgccccggcg gtcatcgcgg 3241 ccgccggcca ggccttctcc cgggtccgcg agggactcgt ggtgccggtc aacccgccgt 3301 ccattcccgg cctgggcacc acgggcggca tggaggtgtg ggtccagaac aagggcgacg 3361 gcagcgtcgg ccaactcgcc gggatagtga acgatttcgt cgccaaggcc cgccagcgac 3421 cggagatcgg cgccatcacc gccaccttca acgccgcctc gcggcaactg ctggccgacg 3481 tggaccggga aaaggcggaa accctgggcg tgccggtgga ggacgtgtac agcaccatgc 3541 agaccatgtt cggctcgttg tacgtgtcgc agttcaacaa gttcagccgc ctgtggcagg 3601 tgatcctgca ggccgagccg acctaccgcc tcagggtcga ggacctgcag cagatctacg 3661 tgcgcaaccg cggcggcgag atgatcccgc tcaaggccct gctgagcacc cgctacgtca 3721 ccggcccgga cctgctgacc cgcttcaaca acttcccggc ggtgaagatc accgccaacg 3781 cggcgccggg cttcagctcc gggcaggcgc tggcggccct ggagacgctg gccgcgcaga 3841 acatgtccgg cgactacgcc ctggccctga gcggcgaggc cttcgaggag aagaaggccg 3901 gcggcaccag cgcccaggtg ttcgtcttcg gcctgatcat ggtgttcctg atcctggcgg 3961 cgcaatacga gaagtggacg ctgccggtcg gcgtgctgct ggcggtgccc ttcgcgctgt 4021 tcggcgccct gctggcgatc ctgttgcgcg gcctggtcaa cgacgtgtac ttccagatcg 4081 gcctgaccat gctggtggcc ctggcggcga agaacgccat cctgatcttc gagttcgccg 4141 tgctgaaccg cgagaaaggc atgtccgtgt tcgacgcggc gatgaccgcg gcgcgcgaac 4201 gcctgcgccc catcgtgatg acctcgctgg ccttcattct cggctgcgtg cccctggcca 4261 tcgccgtcgg cgcctcggag aacagccgcc actcgatcgg taccggggtg atcggcggca 4321 tgctggcggc gacggtgatc gcggtgttct tcatcccgct gttctactac ctgatcgagc 4381 ggctttccga gaagagcggc accgcgaagc aggaagcgcc gcccgccgag ccggcgcccc 4441 tgggcggcga cctgagcggc gtgccgcaac gccggcgcga ggacgaccat catcaccatc 4501 accactgagg atccaaactc gagtaaggat ctccaggcat caaataaaac gaaaggctca 4561 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tctactagag 4621 tcacactggc tcaccttcgg gtgggccttt ctgcgtttat acctagggat atattccgct 4681 tcctcgctca ctgactcgct acgctcggtc gttcgactgc ggcgagcgga aatggcttac 4741 gaacggggcg gagatttcct ggaagatgcc aggaagatac ttaacaggga agtgagaggg 4801 ccgcggcaaa gccgtttttc cataggctcc gcccccctga caagcatcac gaaatctgac 4861 gctcaaatca gtggtggcga aacccgacag gactataaag ataccaggcg tttccccctg 4921 gcggctccct cgtgcgctct cctgttcctg cctttcggtt taccggtgtc attccgctgt 4981 tatggccgcg tttgtctcat tccacgcctg acactcagtt ccgggtaggc agttcgctcc 5041 aagctggact gtatgcacga accccccgtt cagtccgacc gctgcgcctt atccggtaac 5101 tatcgtcttg agtccaaccc ggaaagacat gcaaaagcac cactggcagc agccactggt 5161 aattgattta gaggagttag tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac 5221 aagttttggt gactgcgctc ctccaagcca gttacctcgg ttcaaagagt tggtagctca 5281 gagaaccttc gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc aagagattac 5341 gcgcagacca aaacgatctc aagaagatca tcttattaat cagataaaat atttctagat 5401 ttcagtgcaa tttatctctt caaatgtagc acctgaagtc agccccatac gatataagtt 5461 gttactagtg cttggattct caccaataaa aaacgcccgg cggcaaccga gcgttctgaa 5521 caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct 5581 ctcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc 5641 tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca 5701 agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc 5761 agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag 5821 caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg 5881 gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca 5941 agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat 6001 gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact 6061 ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc 6121 agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc 6181 gtggccagcc acgatagccg cgctgcctcg tcctgcagtt cattcagggc accggacagg 6241 tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca 6301 gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc 6361 ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct 6421 tgatcagatc atgatcccct gcgccatcag atccttggcg gcaagaaagc catccagttt 6481 actttgcagg gcttcccaac cttaccagag ggcgccccag ctggcaattc cgacgtcggt 6541 gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg 6601 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg 6661 cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc 6721 cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag 6781 gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc 6841 gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg 6901 cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc 6961 attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc 7021 cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg 7081 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac 7141 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg 7201 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc 7261 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag 7321 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac 7381 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc 7441 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg 7501 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg 7561 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac 7621 accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg 7681 actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc 7741 cgggatctcg acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt 7801 tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca 7861 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga 7921 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac 7981 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atcgtttagg 8041 caccccaggc tttacacttt atgcttccgg ctcgtataat gtgtggaatt gtgagcggat 8101 aacaatttca gaattcaaaa gatcttttaa gaaggagata tacat //