LOCUS Exported 8142 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pBbA5k-EPL101 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8142) 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 8142) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8142 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 4482..4502 /codon_start=1 /product="6xHis affinity tag" /label=7xHis /translation="HHHHHHH" terminator 4538..4609 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 4625..4652 /label=T7Te terminator /note="phage T7 early transcription terminator" primer_bind complement(4680..4697) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4814..5359) /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(4894..4913) /label=pBR322ori-F /note="pBR322 origin, forward primer" terminator 5473..5567 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(5598..6392) /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(5709..5728) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 6319..6338 /label=Neo-R /note="Neomycin resistance gene, reverse primer" CDS complement(6577..7659) /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 7621..7640 /label=LacI-R /note="LacI, reverse primer" promoter complement(7660..7737) /gene="lacI" /label=lacI promoter primer_bind 7943..7962 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" promoter 8048..8078 /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" protein_bind 8086..8102 /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 atggccggtt catcagtttc aaaaattaag ttgtggatgt tagtttttag cgccacggca 61 atgtcagcct gtggtgatgc gccagcaccg caacaagcta tgctgccaag cgttattgtc 121 aacacagctc atacagagct gatcacgccc aaatctgaaa ttgtcggccg tacacaggca 181 tctgaagatg tgattatcaa gtcgcaaatt catggccagt tgctcaaacg tactttcgtt 241 gaaggtgatg atattaccgc tggcgatcta ttgtttgaaa tcgatccatc tacttataaa 301 gctgagttag cacagcaaaa tgcagtatta aagcaagctg ttgcatcaag ggatgtggcg 361 gtgatgaact gggaacgtgg acgccgatta ttgcctgatg gcatgatcag tgcgcaggat 421 atggatgagc taactagccg taaactcacc actgccgcag gagtcgtaca agccgaagcc 481 gccgtgcaag ctgcagagct gcaattaagt tataccaagg tatatgcgcc aatatcaggc 541 cggatcagtc actcaaaggt gagtaccggt gatatcatta caccacaatc agaaatggca 601 agcctagtgc agcttcagcc tatgtgggta agctttcaag ttgcagaaaa aaagctgatt 661 tcagcccaac aaaacttttc taaggctgtc gaaaaagacg ttaaaattac cgatattgtc 721 atgcgccttc gtctaccgaa cggcaccatg tttgatgagg ttggttatat cgactttatt 781 ggtaaccgag tcgatgccgc aacaggtacc ttgcctgttc gcgctacatt taaaaatgat 841 gacaagctga tgcttccagg cttgtttgtc actctagtga ttgaatcccc cattactgaa 901 aatgcactac ttattcctca agctgcggtg caggaagatc aacaaggccg tttcgtgatg 961 gtgcttaatg atcaagatga agtacagaaa cgtatcgtcg aactgggcga ccgtttcggt 1021 gttgagtggc gtgtgctcag tggtctagag gatggtgaac gcattgtggt ggaggggtta 1081 cagaaaattc gtccaggtat gaaagtgaac ccagtagagc aagaagtggt gcctttcaaa 1141 gaagatgaaa aatcatagtc ggagcagtca agatgaatga gttttttatt agagctgaag 1201 acctaagttt ccccgttttg gctaagcctt tgtataacga ggaatgtcat gatgagtaag 1261 ttttttatta gaacagaagc tctaagtttg ctggttgtga ctaagccttt gttgaacagg 1321 ggaatgtcat gattagtgat ttttttatta ataggcctaa gtttgccttt gttatctcaa 1381 ccgttttgac cttggttggc ttgatcgcta ttccagtgtt atcggtttca gaatttccag 1441 aaattgcgcc acctcaggtg agcgtttcaa ctagttactc tggtgccagc gccgatattg 1501 ttaaagacac catagcgcaa ccattagaag ccgaagttaa cggcgtcgaa ggcatgctgt 1561 atatggagtc aaaaagtgct aacgacggta gctactcgct aaatgtgacc tttgaggttg 1621 gtaccgatgc cgatatggct caggtgaagg tacaaaaccg agtgcagcag gccatgccgc 1681 gtctacctga agaggtgaag cgtcaagggg ttaaagtcga aaaacaaagc cctaacatgc 1741 tgatggtggt caacctagtt tcacctaatg aaacttttga ttctctgttt attactaact 1801 atgctggctt aaacgttaaa gacgcactgg cccgtcagta tggtgtgtct aaggtgcaag 1861 tcattggtgc actggattac gctatgcgta tttggctcga tccagataag atggcaagtc 1921 ttggggtaac agcaacggat gtaattgctg cgctgcaaga acaaaatatt caagtggctg 1981 ctggccgaat tggcgccgca ccagtcgatc cagagcaaca atttcagtac acattgcaga 2041 ccaaaggtcg ccttaaagat cctaaagagt tttatgatgt gatgatccgg gctaataatg 2101 atggctctaa agtcgttgtt ggcgatgtcg cacgagttga gcttggctcc caaacctatg 2161 atgctcaagg taagctaaat aacaaacctt cggcgattat cgctatctat cagtcacccg 2221 atgcgaatgc attagaagtc ggtaaagcca ttaaggccga aatggaaaac ctgtctgagc 2281 gttttccaaa cgatctagaa tatgaagttc tatacgatac aactgagttt gttgaaacct 2341 caattaaaga ggtggtgcaa actctgttta tctctatttc actggtggtc ttggttgtat 2401 ttattttctt acaggacgtg cgttcaacat tagtgcctgc gattgctatt cctgtttcgc 2461 ttatcggtac atttgccttc ttgcttgctt ttgatatgag catcaatacc gtctctctgt 2521 ttgcactgat cctcgcaatt ggtatcgtgg tggacgatgc gattgtagtg gtggagaacg 2581 ttacacgtct gatgcaagat gaaggattat cgccaaaaga ggcgacctct aaggcgatga 2641 aagaggtcac tgggccgatt attgcgacga ctttggtatt gcttgcggta tttgcaccaa 2701 cggcggtcat gccggggatc acagggcaga tgtatgctca gttttctgtg accatctgta 2761 tttctgtact gatctcttca attaacgcac taaccttaag cccagcgctt tgtgcatcat 2821 tactacgccc gcctaagaca cacaccaagg gttttcatgc agtctttaat aagtactttg 2881 aacgatttac tggcaagtat atgaagctag tgtcgttgct gacacgtaag ttagctatcg 2941 tcggtgttgt gtatatttgt ctaattgttg tcactggtgg ggttgccaaa atcttaccgt 3001 cgggctttgt gccgatggaa gacaagaaag cctttatggt ggatattcag ctacccgatg 3061 gcgcatcact taaccgtacc gaagacgtga tgcgagagct ggttgaacta acacttgccg 3121 aacctggtgt ggagaatgtg attcatgcca gtggctttag tattttaacc ggttcagtat 3181 catcaaatgg tggtttgatg attgttactc tgtcgacctg gagtgaacgt gaatctcccg 3241 acatggttga gtcggcgatt gtggctaagt tgcaagctaa atatgcggcc aatcctgcag 3301 tgaaggcgat ggcattctca ctgccgccaa ttccaggggt aggtagtgtt ggtggctttg 3361 agtttgtatt gcaagacact caaggccgta caccacaaga gttggcttct gtgatgcgcg 3421 ccttgattat gaaggccaat gaacaaccag aaattgccat ggcctttagt aacttccgcg 3481 ccgatgtgcc gcaaatgttt gtcgatgtcg acagggataa agctaaagca cttgggatct 3541 cgttaaatga gatcttctcg accatgcaaa ctatgcttgg ctctatgtac gtcaatgatt 3601 ttaaccgttt tggtaaggta tttagggtta tcttgcaggc ggaatctgag tatcgtaatt 3661 ccgacagaga tatctcgcgc ttctacgtcc gctctaacac cggtgagatg gtgccattaa 3721 gtactttggt gacagtgacg ccaattttgg ggcctgacgt gatgaataac tacaacatgt 3781 ttagttccac aactatcaat ggcttccccg ctcccggctt tagttcgggt gatgcaatta 3841 ctgcgatgga gcgtgctgcc aatgaatcac ttccggcagg atataccttc gagtggacag 3901 ggcaaaccta tcaagagatc aaagcgggta atctagcgcc gcttattttc tcgttggcat 3961 tagtgtttac ttacctgttc ttagtggcgc agtacgagag ttggacgatt ccatttgccg 4021 ttattctcgc agtacctatt gcggtactgg gggcatttct caacatcttg ctcgttgggt 4081 cggatttaaa cctctatgcc cagattggtc tggtgctgct tatcggcctc gcctgtaaga 4141 atgccatctt gattgttgag tttgctaaac aattgcacga agagggtaag agtatccttg 4201 acgctgcgca gacggcggca aggctgcgtt ttcgtgcggt gctgatgacg gctttttctt 4261 tcctattggg ggtgttgcct ttagttatcg ctacaggcgc gggggcgggg agtcgccgtg 4321 cccttggtta ctcggtgttt ggcggtatgt tagcggccac tatcgtaggt accctgctgg 4381 taccagtgtt ttacgtaatc atgcaaaaaa tgcgtgaaaa ggcgaaaggc ttgcctaaag 4441 agtccgagca acctcaggag caagagccga aagaagcgag tcaccatcat caccatcacc 4501 actgaggatc caaactcgag taaggatctc caggcatcaa ataaaacgaa aggctcagtc 4561 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca 4621 cactggctca ccttcgggtg ggcctttctg cgtttatacc tagggatata ttccgcttcc 4681 tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa 4741 cggggcggag atttcctgga agatgccagg aagatactta acagggaagt gagagggccg 4801 cggcaaagcc gtttttccat aggctccgcc cccctgacaa gcatcacgaa atctgacgct 4861 caaatcagtg gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggcg 4921 gctccctcgt gcgctctcct gttcctgcct ttcggtttac cggtgtcatt ccgctgttat 4981 ggccgcgttt gtctcattcc acgcctgaca ctcagttccg ggtaggcagt tcgctccaag 5041 ctggactgta tgcacgaacc ccccgttcag tccgaccgct gcgccttatc cggtaactat 5101 cgtcttgagt ccaacccgga aagacatgca aaagcaccac tggcagcagc cactggtaat 5161 tgatttagag gagttagtct tgaagtcatg cgccggttaa ggctaaactg aaaggacaag 5221 ttttggtgac tgcgctcctc caagccagtt acctcggttc aaagagttgg tagctcagag 5281 aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt tcagagcaag agattacgcg 5341 cagaccaaaa cgatctcaag aagatcatct tattaatcag ataaaatatt tctagatttc 5401 agtgcaattt atctcttcaa atgtagcacc tgaagtcagc cccatacgat ataagttgtt 5461 actagtgctt ggattctcac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa 5521 atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa gcgagctctc 5581 gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc 5641 gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc 5701 tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc 5761 cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag 5821 gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgcgcgcctt gagcctggcg 5881 aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga 5941 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg 6001 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc 6061 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc 6121 cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg 6181 gccagccacg atagccgcgc tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg 6241 gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag 6301 cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga 6361 gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga 6421 tcagatcatg atcccctgcg ccatcagatc cttggcggca agaaagccat ccagtttact 6481 ttgcagggct tcccaacctt accagagggc gccccagctg gcaattccga cgtcggtgcc 6541 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga 6601 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 6661 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt 6721 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg 6781 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc 6841 gtatcccact accgagatgt ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat 6901 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt 6961 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc 7021 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc 7081 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag 7141 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt 7201 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat 7261 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag 7321 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac 7381 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg 7441 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg 7501 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt 7561 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc 7621 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact 7681 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg 7741 gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga 7801 ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc 7861 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt 7921 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg 7981 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc gtttaggcac 8041 cccaggcttt acactttatg cttccggctc gtataatgtg tggaattgtg agcggataac 8101 aatttcagaa ttcaaaagat cttttaagaa ggagatatac at //