LOCUS       sequence-167866-        9220 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses efflux pump in E. coli to provide resistance to biofuel 
            (low copy).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9220)
  AUTHORS   Turner WJ, Dunlop MJ
  TITLE     Trade-Offs in Improving Biofuel Tolerance Using Combinations of 
            Efflux Pumps.
  JOURNAL   ACS Synth Biol. 2015 Oct 16;4(10):1056-63. doi: 10.1021/sb500307w. 
            Epub 2014 Dec 12.
  PUBMED    25496359
REFERENCE   2  (bases 1 to 9220)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1021/sb500307w";
            journalName: "ACS Synth Biol"; date: "2015-10-16- 16"; volume: "4"; 
            issue: "10"; pages: "1056-63"
COMMENT     Sequence Label: pBbS5c-PP_3456
FEATURES             Location/Qualifiers
     source          1..9220
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(49..1131)
                     /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     1093..1112
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1132..1209)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1415..1434
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        join(1520..1525,1526..1543,1544..1550)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1558..1574
                     /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)."
     terminator      5963..6034
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6050..6077
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6608..7558)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(8320..8414)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(8435..9094)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     9009..9028
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(9095..9197)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 gacgtcggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
       61 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
      121 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
      181 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
      241 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
      301 ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc cggactcggt
      361 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
      421 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
      481 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
      541 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
      601 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
      661 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
      721 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
      781 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
      841 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
      901 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
      961 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     1021 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     1081 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     1141 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     1201 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     1261 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     1321 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     1381 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     1441 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
     1501 tcgtttaggc accccaggct ttacacttta tgcttccggc tcgtataatg tgtggaattg
     1561 tgagcggata acaatttcag aattcaaaag atcttttaag aaggagatat acatatgcct
     1621 actaccctct ccccgtgcct gcgcccgttg gcgctcatgg ccttgctgag cctgtccctg
     1681 gccggctgca gcgacagcgc cgagccagac gaacaggccc ccaccgcgca agtgcgcgta
     1741 gaaaccctgc aactgcagcc actggccatc agcagtgaac tcagcggccg tatcctcgcg
     1801 ccacgcaccg ccgaagtgcg tgcgcgggtc gcaggcgtgg tgctcaagcg ggtgtaccgc
     1861 gaaggcagcg acgtcaagca gggtgatgta ctgttcctga tcgacccggc gccgttcaag
     1921 gccgaccatg acagcgcccg cgccacactg gccaaggccg aagccacccg gtaccaggcg
     1981 cgcctgcaag agcagcgcta ccgcgagctg gtcgacgaca aagccgtcag ccgccaggaa
     2041 tacgacaacg ccaaggccag tttcctgcag gccgatgccg aagtggccga agccagggcg
     2101 gcgctggagc gcgcccggct gaacctcggc tatgccaccg tgaccgcgcc gatctccggc
     2161 cgtatcggcc gcgcccaggt caccgaaggc gcgctggtcg ggcaaaacga aaccaccccg
     2221 ctggcgacca tccagcagtt ggacccgatc catgccgatg tcacccagtc tacccgcgag
     2281 ctcaacgccc tgcgccgcgc cctgcgcgcc ggcgaattgc agcaggtggg cgacggccag
     2341 gcacgcgcca cgctgatcca ggatgacggc agcgcctacc cgctgccggg caagctgctg
     2401 ttctccgaca tcagcgtcga ccccagcacc aaccagatca ccctgcgcag cgagttcccc
     2461 aacccggacc tcgacctgct gccaggcagc tacgtgcggg tacgcctgga gcaagcggtg
     2521 caacccaagg gcctgagcgt gccgcaacgc gccatcctgc gcgacagcgc cggtgtgccc
     2581 aaggtgctgg tggtagacca gcacgcccgg gtcagcgacc gccaggttgt actgggcagc
     2641 gcccagggtg accgctggat cgtcagcgag ggcctggcgg ccggtgagcg ggtggtggtc
     2701 gagggcctgc agcacgtcaa ggccggtgac caggtgcagg tcgacaacac gcccgctgcc
     2761 ccacccatcg cacagcatac cggccagtga gggcctgatt catgccgcaa ttcttcatcg
     2821 atcgcccggt attcgcctgg gtggtcgcac tgttcatcct gctggctggc gcgctggcca
     2881 tcccgcaact gccggtggcc cagtacccca acgtggcacc gccgcaggtg gaaatctacg
     2941 ccgtgtaccc gggtgcctcg gcggcgacca tggacgaaag cgtggtcagc ctgatcgaac
     3001 aggagctcaa cggcgccgac aacctgctgt acttcgagtc gcagagcagc ctgggcagcg
     3061 ccaccatcac cgccaccttc gccccgggta cccacccgga cctggcccag gtcgacgtgc
     3121 agaaccgcct gaaagtggtc gagtcgcgcc tgccgcgccc ggtcacccag cagggcctgc
     3181 aggtggagaa agtgtccacc ggcttcctgc tgttggcgac cctcacctcc gaagacggca
     3241 agctcgatga aaccgcgctg tcggacatcc ttgcacgtaa cgtgatggac gagatccgcc
     3301 gcctcaaggg cgtcggcaag gcccagttgt atggctccga acgggccatg cgcatctgga
     3361 tcgacccgcg caagctgatc ggtttcaacc tcacgcccaa cgatgtagcc gaagccatcg
     3421 ccgcgcagaa cgcccaggtc gcccctggca gcatcggcga tctgcccagc cgcagcaccc
     3481 aggaaatcac cgccaacgtg gtggtcaaag gccagttgag cagccccgac gagtttgctg
     3541 ccattgtcct gcgtgccaac ccggacggct ccacggtcac catcggggat gtcgcccggg
     3601 ttgaaatcgg tgcccaggaa taccagtacg gcacgcgcct gaacggcaaa ccggccacgg
     3661 ccttcagcgt gcagttgtca cccggcgcca acgccatgga aaccgctacc ctggtacggg
     3721 cgaagatgca ggacctggcg cgctacttcc cggaaggcgt caagtacgac atcccctacg
     3781 acacctcgcc gttcgtcaag gtgtcgatcg agcaggtcat caacaccctg ttcgaggcca
     3841 tgctgctggt gttcgcagtc atgttcctgt tcctgcagaa cctgcgctac acgctgatcc
     3901 cgaccctggt cgtgccagtg gcattgatgg gcaccttcgc cgtgatgctg gccatgggct
     3961 tctcggtcaa tgtgctgaca ctgttcggca tggtgctggc catcggcatt ctggtcgacg
     4021 atgccatcgt ggtggtggag aacgtcgagc gcatcatggc cgaagaaggc ttgccgccca
     4081 agcaagccac gcgcaaggcc atgggccaga tcagcggtgc aatcgtcggc atcacgctgg
     4141 tgctggtggc ggtattcctg cccatggcct tcatgcaagg ctcggtgggg gtgatctacc
     4201 agcagttctc gctgtcgatg gcggtgtcga tcctgttctc ggcgtttctc gccctcagcc
     4261 tgaccccggc attgtgcgcc accctgctca agcccgtggc caagggcgag caccatgaac
     4321 gcaaaggctt cttcggctgg ttcaatcgcc gcttcgaaag catgagcaac ggctaccagc
     4381 gctgggtagt gcaggcgctc aagcgcagcg gccgctacct gctggtgtac gcggtgctgc
     4441 tggcggtgct gggctacggt ttcagccagc tgcctacggc gttcctgcct accgaagacc
     4501 aaggctacac catcaccgac attcagctgc cgccgggcgc cagccgcatg cgcaccgaac
     4561 aggtagccgc gcagatcgag gcgcacaacg ccgaagaacc gggcgtgggc aacaccacct
     4621 tgatcctcgg cttcagcttc tccggtagcg ggcagaacgc ggcgctggcc ttcaccaccc
     4681 tcaaggactg gtccgagcgc ggcgccgacg acagcgccca gtcgatcgcc gaccgcgcga
     4741 ccatggcctt cacccagctc aaggacgcga ttgcctattc ggtactgccg ccccccattg
     4801 acggcctcgg tgagtcgacc ggtttcgaat ttcgccttca ggaccgtggc ggcatgggcc
     4861 acgccgagct gatggcagcc cgtgaccagc tgctcgaaag cgccagcaaa agcaaggtgc
     4921 tgaccaacgt gcgcgaagcc tcgctggcgg aaagcccgca ggtgcaactg gaaatcgacc
     4981 gccgccaggc taatgcactg ggtgtgtcgt ttgccgatat cggcacggtg ctggacgtgg
     5041 ccgtgggctc cagctacgtc aacgatttcc ccaaccaggg ccgcatgcag cgggtggtgg
     5101 tgcaggccga aggtgaccag cgcagccagg tcgaagactt gctgaacatt cacgtgcgca
     5161 acgacagcgg caagatggtg ccactgggcg cgttcgtgca ggccaggtgg gtcagcggcc
     5221 cggtgcaact gacccgctac aacggttacc cggcggtgtc gatctcaggc gagcctgccg
     5281 ccggctacag ctcgggcgag gccatggccg aagtcgagcg gctggtggcg cagctaccgg
     5341 ccggtaccgg tctggaatgg accggcctgt cactgcagga gcggctgtcc ggtagccagg
     5401 cccccctgct gatggcgctg tcgctgctgg tggtgttcct gtgcctggca gcgctgtacg
     5461 aaagctggtc gatcccgact gcggtgctgc tggtcgttcc gctcggtgtg ctcggtgcgg
     5521 tgctggcggt gaccttgcgc ggcatgccca acgacgtgtt cttcaaggtt ggcctgatta
     5581 ccctgatcgg cctgtcggcg aagaacgcca tcctgatcat cgagttcgcc aagcatctgg
     5641 ttgaccaggg cgtggacgcc gccgacgccg ccgtgcaagc ggcccgcctg cgcctgcggc
     5701 cgatcgtgat gacgtcgctg gcgttcatcc tcggcgtggt gcccctggcc atcgccagcg
     5761 gcgcaagctc tgccagccag caggcgattg gcaccggggt gatcggcggc atgctcagcg
     5821 ccaccctggc ggtggtgttc gtgccggtgt tcttcgtggt ggtgatgcgc ctgtccgggc
     5881 gacgccaggc gcatgacagc gacgggcaac ccgtgccccg cgaaagctga ggatccaaac
     5941 tcgagtaagg atctccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
     6001 tcgttttatc tgttgtttgt cggtgaacgc tctctactag agtcacactg gctcaccttc
     6061 gggtgggcct ttctgcgttt atacctaggg tacgggtttt gctgcccgca aacgggctgt
     6121 tctggtgttg ctagtttgtt atcagaatcg cagatccggc ttcagccggt ttgccggctg
     6181 aaagcgctat ttcttccaga attgccatga ttttttcccc acgggaggcg tcactggctc
     6241 ccgtgttgtc ggcagctttg attcgataag cagcatcgcc tgtttcaggc tgtctatgtg
     6301 tgactgttga gctgtaacaa gttgtctcag gtgttcaatt tcatgttcta gttgctttgt
     6361 tttactggtt tcacctgttc tattaggtgt tacatgctgt tcatctgtta cattgtcgat
     6421 ctgttcatgg tgaacagctt tgaatgcacc aaaaactcgt aaaagctctg atgtatctat
     6481 cttttttaca ccgttttcat ctgtgcatat ggacagtttt ccctttgata tgtaacggtg
     6541 aacagttgtt ctacttttgt ttgttagtct tgatgcttca ctgatagata caagagccat
     6601 aagaacctca gatccttccg tatttagcca gtatgttctc tagtgtggtt cgttgttttt
     6661 gcgtgagcca tgagaacgaa ccattgagat catacttact ttgcatgtca ctcaaaaatt
     6721 ttgcctcaaa actggtgagc tgaatttttg cagttaaagc atcgtgtagt gtttttctta
     6781 gtccgttatg taggtaggaa tctgatgtaa tggttgttgg tattttgtca ccattcattt
     6841 ttatctggtt gttctcaagt tcggttacga gatccatttg tctatctagt tcaacttgga
     6901 aaatcaacgt atcagtcggg cggcctcgct tatcaaccac caatttcata ttgctgtaag
     6961 tgtttaaatc tttacttatt ggtttcaaaa cccattggtt aagcctttta aactcatggt
     7021 agttattttc aagcattaac atgaacttaa attcatcaag gctaatctct atatttgcct
     7081 tgtgagtttt cttttgtgtt agttctttta ataaccactc ataaatcctc atagagtatt
     7141 tgttttcaaa agacttaaca tgttccagat tatattttat gaattttttt aactggaaaa
     7201 gataaggcaa tatctcttca ctaaaaacta attctaattt ttcgcttgag aacttggcat
     7261 agtttgtcca ctggaaaatc tcaaagcctt taaccaaagg attcctgatt tccacagttc
     7321 tcgtcatcag ctctctggtt gctttagcta atacaccata agcattttcc ctactgatgt
     7381 tcatcatctg agcgtattgg ttataagtga acgataccgt ccgttctttc cttgtagggt
     7441 tttcaatcgt ggggttgagt agtgccacac agcataaaat tagcttggtt tcatgctccg
     7501 ttaagtcata gcgactaatc gctagttcat ttgctttgaa aacaactaat tcagacatac
     7561 atctcaattg gtctaggtga ttttaatcac tataccaatt gagatgggct agtcaatgat
     7621 aattactagt ccttttcccg ggtgatctgg gtatctgtaa attctgctag acctttgctg
     7681 gaaaacttgt aaattctgct agaccctctg taaattccgc tagacctttg tgtgtttttt
     7741 ttgtttatat tcaagtggtt ataatttata gaataaagaa agaataaaaa aagataaaaa
     7801 gaatagatcc cagccctgtg tataactcac tactttagtc agttccgcag tattacaaaa
     7861 ggatgtcgca aacgctgttt gctcctctac aaaacagacc ttaaaaccct aaaggcttaa
     7921 gtagcaccct cgcaagctcg ggcaaatcgc tgaatattcc ttttgtctcc gaccatcagg
     7981 cacctgagtc gctgtctttt tcgtgacatt cagttcgctg cgctcacggc tctggcagtg
     8041 aatgggggta aatggcacta caggcgcctt ttatggattc atgcaaggaa actacccata
     8101 atacaagaaa agcccgtcac gggcttctca gggcgtttta tggcgggtct gctatgtggt
     8161 gctatctgac tttttgctgt tcagcagttc ctgccctctg attttccagt ctgaccactt
     8221 cggattatcc cgtgacaggt cattcagact ggctaatgca cccagtaagg cagcggtatc
     8281 atcaacaggc ttacccgtct tactgtccct agtgcttgga ttctcaccaa taaaaaacgc
     8341 ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc
     8401 tatcaacagg agtccaagcg agctcgatat caaattacgc cccgccctgc cactcatcgc
     8461 agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat
     8521 gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg
     8581 tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac
     8641 tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg
     8701 ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat
     8761 cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt
     8821 aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc atacgaaatt
     8881 ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct
     8941 tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg
     9001 tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat
     9061 caacggtggt atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc
     9121 tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac
     9181 ctcttacgtg ccgatcaacg tctcattttc gccagatatc
//
