LOCUS       sequence-383914-        8751 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression vector for transcriptionally fused acrAB and 
            sfGFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8751)
  AUTHORS   Wen X, Langevin AM, Dunlop MJ
  TITLE     Antibiotic export by efflux pumps affects growth of neighboring 
            bacteria.
  JOURNAL   Sci Rep. 2018 Oct 11;8(1):15120. doi: 10.1038/s41598-018-33275-4.
  PUBMED    30310093
REFERENCE   2  (bases 1 to 8751)
  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"; journalName: "Sci Rep."; 
            date: "2018-10-11"; volume: "8(1):15120. doi"; pages: " 
            10.1038/s41598-018-33275-4"
COMMENT     Sequence Label: pBbA5k-acrAB-sfGFP
FEATURES             Location/Qualifiers
     source          1..8751
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(21..26,27..44,45..51)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    59..75
                     /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)."
     RBS             4487..4498
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4507..5220
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      5262..5333
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5349..5376
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(5404..5421)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5538..6083)
                     /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(5618..5637)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(6197..6291)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(6322..7116)
                     /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(6433..6452)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7043..7062
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(7301..8383)
                     /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     8345..8364
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(8384..8461)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     8667..8686
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atcgtttagg caccccaggc tttacacttt atgcttccgg ctcgtataat gtgtggaatt
       61 gtgagcggat aacaatttca gaattcaaaa gatcttttaa gaaggagata tacatatgaa
      121 caaaaacaga gggtttacgc ctctggcggt cgttctgatg ctctcaggca gcttagccct
      181 aacaggatgt gacgacaaac aggcccaaca aggtggccag cagatgcccg ccgttggcgt
      241 agtaacagtc aaaactgaac ctctgcagat cacaaccgag cttccgggtc gcaccagtgc
      301 ctaccggatc gcagaagttc gtcctcaagt tagcgggatt atcctgaagc gtaatttcaa
      361 agaaggtagc gacatcgaag caggtgtctc tctctatcag attgatcctg cgacctatca
      421 ggcgacatac gacagtgcga aaggtgatct ggcgaaagcc caggctgcag ccaatatcgc
      481 gcaattgacg gtgaatcgtt atcagaaact gctcggtact cagtacatca gtaagcaaga
      541 gtacgatcag gctctggctg atgcgcaaca ggcgaatgct gcggtaactg cggcgaaagc
      601 tgccgttgaa actgcgcgga tcaatctggc ttacaccaaa gtcacctctc cgattagcgg
      661 tcgcattggt aagtcgaacg tgacggaagg cgcattggta cagaacggtc aggcgactgc
      721 gctggcaacc gtgcagcaac ttgatccgat ctacgttgat gtgacccagt ccagcaacga
      781 cttcctgcgc ctgaaacagg aactggcgaa tggcacgctg aaacaagaga acggcaaagc
      841 caaagtgtca ctgatcacca gtgacggcat taagttcccg caggacggta cgctggaatt
      901 ctctgacgtt accgttgatc agaccactgg gtctatcacc ctacgcgcta tcttcccgaa
      961 cccggatcac actctgctgc cgggtatgtt cgtgcgcgca cgtctggaag aagggcttaa
     1021 tccaaacgct attttagtcc cgcaacaggg cgtaacccgt acgccgcgtg gcgatgccac
     1081 cgtactggta gttggcgcgg atgacaaagt ggaaacccgt ccgatcgttg caagccaggc
     1141 tattggcgat aagtggctgg tgacagaagg tctgaaagca ggcgatcgcg tagtaataag
     1201 tgggctgcag aaagtgcgtc ctggtgtcca ggtaaaagca caagaagtta ccgctgataa
     1261 taaccagcaa gccgcaagcg gtgctcagcc tgaacagtcc aagtcttaac ttaaacagga
     1321 gccgttaaga catgcctaat ttctttatcg atcgcccgat ttttgcgtgg gtgatcgcca
     1381 ttatcatcat gttggcaggg gggctggcga tcctcaaact gccggtggcg caatatccta
     1441 cgattgcacc gccggcagta acgatctccg cctcctaccc cggcgctgat gcgaaaacag
     1501 tgcaggacac ggtgacacag gttatcgaac agaatatgaa cggtatcgat aacctgatgt
     1561 acatgtcctc taacagtgac tccacgggta ccgtgcagat caccctgacc tttgagtctg
     1621 gtactgatgc ggatatcgcg caggttcagg tgcagaacaa actgcagctg gcgatgccgt
     1681 tgctgccgca agaagttcag cagcaagggg tgagcgttga gaaatcatcc agcagcttcc
     1741 tgatggttgt cggcgttatc aacaccgatg gcaccatgac gcaggaggat atctccgact
     1801 acgtggcggc gaatatgaaa gatgccatca gccgtacgtc gggcgtgggt gatgttcagt
     1861 tgttcggttc acagtacgcg atgcgtatct ggatgaaccc gaatgagctg aacaaattcc
     1921 agctaacgcc ggttgatgtc attaccgcca tcaaagcgca gaacgcccag gttgcggcgg
     1981 gtcagctcgg tggtacgccg ccggtgaaag gccaacagct taacgcctct attattgctc
     2041 agacgcgtct gacctctact gaagagttcg gcaaaatcct gctgaaagtg aatcaggatg
     2101 gttcccgcgt gctgctgcgt gacgtcgcga agattgagct gggtggtgag aactacgaca
     2161 tcatcgcaga gtttaacggc caaccggctt ccggtctggg gatcaagctg gcgaccggtg
     2221 caaacgcgct ggataccgct gcggcaatcc gtgctgaact ggcgaagatg gaaccgttct
     2281 tcccgtcggg tctgaaaatt gtttacccat acgacaccac gccgttcgtg aaaatctcta
     2341 ttcacgaagt ggttaaaacg ctggtcgaag cgatcatcct cgtgttcctg gttatgtatc
     2401 tgttcctgca gaacttccgc gcgacgttga ttccgaccat tgccgtaccg gtggtattgc
     2461 tcgggacctt tgccgtcctt gccgcctttg gcttctcgat aaacacgcta acaatgttcg
     2521 ggatggtgct cgccatcggc ctgttggtgg atgacgccat cgttgtggta gaaaacgttg
     2581 agcgtgttat ggcggaagaa ggtttgccgc caaaagaagc tacccgtaag tcgatggggc
     2641 agattcaggg cgctctggtc ggtatcgcga tggtactgtc ggcggtattc gtaccgatgg
     2701 ccttctttgg cggttctact ggtgctatct atcgtcagtt ctctattacc attgtttcag
     2761 caatggcgct gtcggtactg gtggcgttga tcctgactcc agctctttgt gccaccatgc
     2821 tgaaaccgat tgccaaaggc gatcacgggg aaggtaaaaa aggcttcttc ggctggttta
     2881 accgcatgtt cgagaagagc acgcaccact acaccgacag cgtaggcggt attctgcgca
     2941 gtacggggcg ttacctggtg ctgtatctga tcatcgtggt cggcatggcc tatctgttcg
     3001 tgcgtctgcc aagctccttc ttgccagatg aggaccaggg cgtgtttatg accatggttc
     3061 agctgccagc aggtgcaacg caggaacgta cacagaaagt gctcaatgag gtaacgcatt
     3121 actatctgac caaagaaaag aacaacgttg agtcggtgtt cgccgttaac ggcttcggct
     3181 ttgcgggacg tggtcagaat accggtattg cgttcgtttc cttgaaggac tgggccgatc
     3241 gtccgggcga agaaaacaaa gttgaagcga ttaccatgcg tgcaacacgc gctttctcgc
     3301 aaatcaaaga tgcgatggtt ttcgccttta acctgcccgc aatcgtggaa ctgggtactg
     3361 caaccggctt tgactttgag ctgattgacc aggctggcct tggtcacgaa aaactgactc
     3421 aggcgcgtaa ccagttgctt gcagaagcag cgaagcaccc tgatatgttg accagcgtac
     3481 gtccaaacgg tctggaagat accccgcagt ttaagattga tatcgaccag gaaaaagcgc
     3541 aggcgctggg tgtttctatc aacgacatta acaccactct gggcgctgca tggggcggca
     3601 gctatgtgaa cgactttatc gaccgcggtc gtgtgaagaa agtttatgtc atgtcagaag
     3661 cgaaataccg tatgctgccg gatgatatcg gcgactggta tgttcgtgct gctgatggtc
     3721 agatggtgcc attctcggcg ttctcctctt ctcgttggga gtacggttcg ccgcgtctgg
     3781 aacgttacaa cggcctgcca tccatggaaa tcttaggcca ggcggcaccg ggtaaaagta
     3841 ccggtgaagc aatggagctg atggaacaac tggcgagcaa actgcctacc ggtgttggct
     3901 atgactggac ggggatgtcc tatcaggaac gtctctccgg caaccaggca ccttcactgt
     3961 acgcgatttc gttgattgtc gtgttcctgt gtctggcggc gctgtacgag agctggtcga
     4021 ttccgttctc cgttatgctg gtcgttccgc tgggggttat cggtgcgttg ctggctgcca
     4081 ccttccgtgg cctgaccaat gacgtttact tccaggtagg cctgctcaca accattgggt
     4141 tgtcggcgaa gaacgcgatc cttatcgtcg aattcgccaa agacttgatg gataaagaag
     4201 gtaaaggtct gattgaagcg acgcttgatg cggtgcggat gcgtttacgt ccgatcctga
     4261 tgacctcgct ggcgtttatc ctcggcgtta tgccgctggt tatcagtact ggtgctggtt
     4321 ccggcgcgca gaacgcagta ggtaccggtg taatgggcgg gatggtgacc gcaacggtac
     4381 tggcaatctt cttcgttccg gtattctttg tggtggttcg ccgccgcttt agccgcaaga
     4441 atgaagatat cgagcacagc catactgtcg atcatcattt aactttaaag aggagaaagg
     4501 taccgcatgc gtaaaggcga agagctgttc actggtgtcg tccctattct ggtggaactg
     4561 gatggtgatg tcaacggtca taagttttcc gtgcgtggcg agggtgaagg tgacgcaact
     4621 aatggtaaac tgacgctgaa gttcatctgt actactggta aactgccggt accttggccg
     4681 actctggtaa cgacgctgac ttatggtgtt cagtgctttg ctcgttatcc ggaccatatg
     4741 aagcagcatg acttcttcaa gtccgccatg ccggaaggct atgtgcagga acgcacgatt
     4801 tcctttaagg atgacggcac gtacaaaacg cgtgcggaag tgaaatttga aggcgatacc
     4861 ctggtaaacc gcattgagct gaaaggcatt gactttaaag aagacggcaa tatcctgggc
     4921 cataagctgg aatacaattt taacagccac aatgtttaca tcaccgccga taaacaaaaa
     4981 aatggcatta aagcgaattt taaaattcgc cacaacgtgg aggatggcag cgtgcagctg
     5041 gctgatcact accagcaaaa cactccaatc ggtgatggtc ctgttctgct gccagacaat
     5101 cactatctga gcacgcaaag cgttctgtct aaagatccga acgagaaacg cgatcatatg
     5161 gttctgctgg agttcgtaac cgcagcgggc atcacgcatg gtatggatga actgtacaaa
     5221 taatagtgag gatccaaact cgagtaagga tctccaggca tcaaataaaa cgaaaggctc
     5281 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctctactaga
     5341 gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta tacctaggga tatattccgc
     5401 ttcctcgctc actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta
     5461 cgaacggggc ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg
     5521 gccgcggcaa agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga
     5581 cgctcaaatc agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5641 ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg
     5701 ttatggccgc gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc
     5761 caagctggac tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa
     5821 ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg
     5881 taattgattt agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga
     5941 caagttttgg tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc
     6001 agagaacctt cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta
     6061 cgcgcagacc aaaacgatct caagaagatc atcttattaa tcagataaaa tatttctaga
     6121 tttcagtgca atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt
     6181 tgttactagt gcttggattc tcaccaataa aaaacgcccg gcggcaaccg agcgttctga
     6241 acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt ccaagcgagc
     6301 tctcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg
     6361 ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc
     6421 aagctcttca gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc
     6481 cagccggcca cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa
     6541 gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg ggcatgcgcg ccttgagcct
     6601 ggcgaacagt tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac
     6661 aagaccggct tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa
     6721 tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac
     6781 tttctcggca ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag
     6841 cagccagtcc cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt
     6901 cgtggccagc cacgatagcc gcgctgcctc gtcctgcagt tcattcaggg caccggacag
     6961 gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc
     7021 agagcagccg attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc
     7081 cggagaacct gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc
     7141 ttgatcagat catgatcccc tgcgccatca gatccttggc ggcaagaaag ccatccagtt
     7201 tactttgcag ggcttcccaa ccttaccaga gggcgcccca gctggcaatt ccgacgtcgg
     7261 tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc
     7321 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     7381 gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc
     7441 ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca
     7501 ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat
     7561 cgtcgtatcc cactaccgag atgtccgcac caacgcgcag cccggactcg gtaatggcgc
     7621 gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct
     7681 cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt
     7741 ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac
     7801 gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga
     7861 ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg
     7921 gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag
     7981 caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga
     8041 gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca
     8101 ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg
     8161 cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt
     8221 gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc
     8281 gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga
     8341 caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt
     8401 gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt
     8461 ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg
     8521 ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac
     8581 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     8641 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     8701 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga g
//
