LOCUS Exported 7893 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pBbA5k-EPL37 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7893) 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 7893) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7893 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 4233..4253 /codon_start=1 /product="6xHis affinity tag" /label=7xHis /translation="HHHHHHH" terminator 4289..4360 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 4376..4403 /label=T7Te terminator /note="phage T7 early transcription terminator" primer_bind complement(4431..4448) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4565..5110) /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(4645..4664) /label=pBR322ori-F /note="pBR322 origin, forward primer" terminator 5224..5318 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(5349..6143) /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(5460..5479) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 6070..6089 /label=Neo-R /note="Neomycin resistance gene, reverse primer" CDS complement(6328..7410) /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 7372..7391 /label=LacI-R /note="LacI, reverse primer" promoter complement(7411..7488) /gene="lacI" /label=lacI promoter primer_bind 7694..7713 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" promoter 7799..7829 /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" protein_bind 7837..7853 /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 atgttgctcc gccgcatgtt gatcatgctc gccgcggtga tcgccgtggt ggcgattctc 61 gccggctaca aggtctactc catccgtcag cagatcgccc ttttcagcgc accgaaaccg 121 ccgatcagcg tgaccgccag cctggccgaa aagcgtccct ggcagagccg cctgccagcc 181 atcggcagcc tcaaggcatt ccagggcgtg accctcaccg ccgaagtctc cggcacggta 241 cgcgacgtac tgttcctttc cggcgaccag gtgaagctgg accaaccgct gatccagttg 301 gaaagcgacg tcgaggaagc caccctgcgc actgccgagg ccgatctcgg cctggccagg 361 gccgagtacc agcgcggccg cgaactgatc ggcagcaagg ccatctcgaa aagcgaattc 421 gatcgtctcg ccgcgcagtg ggccaagacc agcgccaccg tcgccgagct gaaggcggcg 481 ctggcgaaga agcgcgtgct cgcgcccttc gccgggacca tcggcatccg ccaggtggac 541 gtcggcgact acgtctcgcc cgggacgccg atcgccacct tgcaggacct ttccaccctg 601 ctcctggatt tccacctgcc cgagcaggac ttccccctgc tcagccgcgg ccaactggtg 661 aaggtccggg tcgccgccta ccccgcccag gtgttcgacg ccgagatcgc cgccatcaac 721 cccaaggtcg acaacgagac ccgcaacctg caggtccgcg ctgccctgga gaacccggac 781 ggcaagctgc tgccgggcat gttcgccaac ctcgaggtga tgttgcctgg cgaggaacaa 841 cgcgtcgtgg tcccggagac ggcgatcacc ttcaccctct acggcgactc gatctacgtc 901 gtcgggcaga agaaggacga gcagggccag gtgtcgaagg atgacaaggg ccagccgcaa 961 caggtcgtcg agcgccgctt cgtcaggatc ggcgaacgcc gcgaaggcct ggcggtggtg 1021 ctcgaaggcc ttgagggcgg cgagcaggta gtgacttccg ggcaactgaa gctcgacaac 1081 ggcgccgcgg tggccatcgt cgccgagcgg gacctccagc aagagcactg agtcgcgcgc 1141 cttcccgctc cgtggcggaa ggcttgccaa gggactgaaa catggctttt accgatccgt 1201 tcatccgtcg tccggtcctg gcgagcgtgg tcagcctgct gatcgtcctg ctcggcatgc 1261 aggccttcag caagctggtg atccgcgagt atccgcaaat ggagaacgcg ctgatcacgg 1321 tgaccacgct ctacgccggc gccaacgcgg aaaccatcca gggctacatc acccagccgc 1381 tgcagcagag cctggccagc gccgaaggca tcgactacat gacctcggtg agccggcaga 1441 actattcgac catctccatc tacgcgcgga tcggcgccaa taccgatcgc ctggtcaccg 1501 agctgctggc caagtccaac gaagtgaaga gccagctgcc gccggacgcc gaggacccgg 1561 tgctgcagaa ggaggccgcg gacgcctcgg cgctgatgta catcagcttc tacagcgagc 1621 agatgaacaa cccgcagatc accgactacc tgtcgcgggt gatccagccc aagctggcga 1681 ccctgcccgg tatcgccgag gcggagatcc tcggcaacca ggtgttcgcc atgcgcctgt 1741 ggctggaccc ggtgaagatg gccgcgttcg gcgtcaccgc cggcgagatc aaccaggcgg 1801 tgcagcagta caacttcctc gccgccgccg gcgaggtgaa gggccagttg gtggtcacca 1861 gcgtcaatgc ttccaccgac ctcaagtcgc cccaggcctt cgccgccatc ccggtgaaga 1921 ccgacggcga ccgccgggtg ctgatgggtg atgtcgcacg ggtcgaactg ggcgccgcca 1981 gctacgacgc gatcagttcg ttcaatggga ttccctcggt ctacatcggc atcaagggca 2041 cgcccagcgc caacccgctg gacgtgatca aggaagtgcg ggcgaagatg cccgaactgg 2101 aagagcaatt gccgcccaac ctcaaggtgt ccatcgccta cgacgccacg cgcttcattc 2161 aggcctccat cgatgaagtg gtgaagaccc tcggcgaggc ggtgctgatc gtcatcgtgg 2221 tggtgttcct gttcctcggc gcgttccgtt cggtactgat cccggtggtg accattccgc 2281 tgtcgatgat cggcgtattg ttcttcatgc aggccatggg ctactcgatc aacctgctga 2341 ccctgctggc gatggtcctg gccatcggcc tggtggtgga cgacgcgatc gtggtggtgg 2401 aaaacatcca ccgccacatc gaggagggca agccgccctt cgaggctgcc ctggagggcg 2461 cgcgggagat cgccgtaccg gtggtcagca tgaccatcac cctcgccgcg gtctacgcgc 2521 cgatcggctt cctcaccggc ctcaccggcg ccctgttcaa ggagttcgcc ttcaccctgg 2581 ccggcgcggt gatcatttcc gggatcgtcg ccctgaccct gtcgccgatg atgtgctcgc 2641 gcctgttgcg ccacgaggag aatccctcgg gcctggcgca tcgcctcgac ctgatcttcg 2701 agggcctgaa gcaacgctac cagcgcgccc tccacggcac cctggacacc cgtccggtgg 2761 tcctggtgtt cgccgtgctg gtactggcgc tgatcccggt actgctgatg ttcaccaaga 2821 aggagctggc gccggaagag gaccagggca tcgtgttcct catgaccaac tcgccgcaaa 2881 ccgccaacct cgactacctg aatcgctaca ccgccgagtt cgaaggcatc ttccgcagct 2941 tcccggagta ttactcggcg ttccagatca acggctacaa cggcgtccag gccggcatcg 3001 gcggcatgct gctcaagccc tgggacgaac gcgaaaagag ccagatggaa ttgctccatg 3061 cggtgcaggc gaagctcaac gagattcccg gcgtgcagat cttcgccttc aacctgccgt 3121 cgctaccggg caccggcgag ggcctgccat tccagttcgt gctcaacacc gccaacgact 3181 acgagtcgct gctgcaagtg gcgcaacggg tgaagcagcg cgccagcgag tccggcaagt 3241 tcgccttcct cgatctcgac ctggcgttcg acaaacccga gctggtggtc gacatcgacc 3301 gcgagaaggc cgcgcagatg ggcgtctcca tgcaggacct gggggtggcg ctggccagcc 3361 tgctcggcga gggcgagatc aaccgcttca ccatcgacgg gcgcagctac aaggtcatcg 3421 cccaggtcga acgcccctat cgcgacaatc ccggctggct cggcagctac tacgtgaaga 3481 gccgcaacgg ccaactggtc gccctctcga ccctgatcga gacccacgaa cgggcacggc 3541 cacggcaact gaaccagttc cagcagctca actcggcgat catctccggc ttccccatcg 3601 tcagcatggg cgaggccatc gagacggttc agcagatcgc ccgcgaggaa gccccgcgtg 3661 gcttcgccgt cgactacgcc ggcgcttccc ggcaatacgt gcaggaaggc agcgcattgc 3721 tggtcacctt cggactggcc ctggcgatca tcttcctggt cctcgccgca cagttcgaaa 3781 gcttccgcga tccgctggtg atcatggtca ccgtgccgct gtcgatctgc ggtgcgctga 3841 taccgctgtt cctcggcgtg tcgagcctga acatctatac ccaggtcggc ctggtgaccc 3901 tgatcggcct gatcagcaaa cacggcatcc tgatcgtcga gttcgccaac cagttgcgcc 3961 acgaacaggg gctggggcgt cgcgaggcca tcgagcaggc cgccgcgatc cgcctgcggc 4021 cagtactgat gaccaccgcg gcgatggtgc tcggggtgat cccgctgatc ctcgccaccg 4081 gcgccggggc ggtcagtcgc ttcgacatcg gcatcgtgat cgcgaccggc atgtcggtgg 4141 gcaccctctt caccctgttc gtcctgccgt gcatctacac cctggtggcc aggccggatg 4201 ccccgccagg cgtgacgcaa gcggcaaacg cacaccatca tcaccatcac cactgaggat 4261 ccaaactcga gtaaggatct ccaggcatca aataaaacga aaggctcagt cgaaagactg 4321 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc tactagagtc acactggctc 4381 accttcgggt gggcctttct gcgtttatac ctagggatat attccgcttc ctcgctcact 4441 gactcgctac gctcggtcgt tcgactgcgg cgagcggaaa tggcttacga acggggcgga 4501 gatttcctgg aagatgccag gaagatactt aacagggaag tgagagggcc gcggcaaagc 4561 cgtttttcca taggctccgc ccccctgaca agcatcacga aatctgacgc tcaaatcagt 4621 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctggc ggctccctcg 4681 tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat tccgctgtta tggccgcgtt 4741 tgtctcattc cacgcctgac actcagttcc gggtaggcag ttcgctccaa gctggactgt 4801 atgcacgaac cccccgttca gtccgaccgc tgcgccttat ccggtaacta tcgtcttgag 4861 tccaacccgg aaagacatgc aaaagcacca ctggcagcag ccactggtaa ttgatttaga 4921 ggagttagtc ttgaagtcat gcgccggtta aggctaaact gaaaggacaa gttttggtga 4981 ctgcgctcct ccaagccagt tacctcggtt caaagagttg gtagctcaga gaaccttcga 5041 aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa gagattacgc gcagaccaaa 5101 acgatctcaa gaagatcatc ttattaatca gataaaatat ttctagattt cagtgcaatt 5161 tatctcttca aatgtagcac ctgaagtcag ccccatacga tataagttgt tactagtgct 5221 tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg 5281 gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctct cgaaccccag 5341 agtcccgctc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga 5401 gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca 5461 atatcacggg tagccaacgc tatgtcctga tagcggtccg ccacacccag ccggccacag 5521 tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca 5581 tgggtcacga cgagatcctc gccgtcgggc atgcgcgcct tgagcctggc gaacagttcg 5641 gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc 5701 atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc 5761 ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga 5821 gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt 5881 cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac 5941 gatagccgcg ctgcctcgtc ctgcagttca ttcagggcac cggacaggtc ggtcttgaca 6001 aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt 6061 gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg 6121 tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcagatcat 6181 gatcccctgc gccatcagat ccttggcggc aagaaagcca tccagtttac tttgcagggc 6241 ttcccaacct taccagaggg cgccccagct ggcaattccg acgtcggtgc ctaatgagtg 6301 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg 6361 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc 6421 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg 6481 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg 6541 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac 6601 taccgagatg tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag 6661 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg 6721 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg 6781 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga 6841 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac 6901 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga 6961 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg 7021 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac 7081 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc 7141 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag 7201 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg 7261 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga 7321 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc 7381 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg 7441 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac 7501 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga 7561 gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca 7621 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc 7681 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg 7741 tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat cgtttaggca ccccaggctt 7801 tacactttat gcttccggct cgtataatgt gtggaattgt gagcggataa caatttcaga 7861 attcaaaaga tcttttaaga aggagatata cat //