LOCUS       sequence-238388-        5912 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  plasmid reporting the expression from the acrAB and mutS promoters.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5912)
  AUTHORS   El Meouche I, Dunlop MJ
  TITLE     Heterogeneity in efflux pump expression predisposes 
            antibiotic-resistant cells to mutation.
  JOURNAL   Science. 2018 Nov 9;362(6415):686-690. doi: 10.1126/science.aar7981.
  PUBMED    30409883
REFERENCE   2  (bases 1 to 5912)
  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: "10.1126/science"; 
            journalName: "Science"; date: "2018-11-9- 9"; volume: "362"; issue: 
            "6415"; pages: "686-690"
COMMENT     Sequence Label: PacrAB-rfp + PmutS-yfp
FEATURES             Location/Qualifiers
     source          1..5912
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      33..104
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      120..147
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(678..1628)
                     /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(2390..2484)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2515..3309)
                     /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(2626..2645)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3236..3255
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     RBS             4401..4412
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      complement(4874..4905)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     RBS             5047..5058
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             5067..5780
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(5097..5125)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5720..5740
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      5813..5899
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggatccaaac tcgagtaagg atctccaggc atcaaataaa acgaaaggct cagtcgaaag
       61 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag agtcacactg
      121 gctcaccttc gggtgggcct ttctgcgttt atacctaggg tacgggtttt gctgcccgca
      181 aacgggctgt tctggtgttg ctagtttgtt atcagaatcg cagatccggc ttcagccggt
      241 ttgccggctg aaagcgctat ttcttccaga attgccatga ttttttcccc acgggaggcg
      301 tcactggctc ccgtgttgtc ggcagctttg attcgataag cagcatcgcc tgtttcaggc
      361 tgtctatgtg tgactgttga gctgtaacaa gttgtctcag gtgttcaatt tcatgttcta
      421 gttgctttgt tttactggtt tcacctgttc tattaggtgt tacatgctgt tcatctgtta
      481 cattgtcgat ctgttcatgg tgaacagctt tgaatgcacc aaaaactcgt aaaagctctg
      541 atgtatctat cttttttaca ccgttttcat ctgtgcatat ggacagtttt ccctttgata
      601 tgtaacggtg aacagttgtt ctacttttgt ttgttagtct tgatgcttca ctgatagata
      661 caagagccat aagaacctca gatccttccg tatttagcca gtatgttctc tagtgtggtt
      721 cgttgttttt gcgtgagcca tgagaacgaa ccattgagat catacttact ttgcatgtca
      781 ctcaaaaatt ttgcctcaaa actggtgagc tgaatttttg cagttaaagc atcgtgtagt
      841 gtttttctta gtccgttatg taggtaggaa tctgatgtaa tggttgttgg tattttgtca
      901 ccattcattt ttatctggtt gttctcaagt tcggttacga gatccatttg tctatctagt
      961 tcaacttgga aaatcaacgt atcagtcggg cggcctcgct tatcaaccac caatttcata
     1021 ttgctgtaag tgtttaaatc tttacttatt ggtttcaaaa cccattggtt aagcctttta
     1081 aactcatggt agttattttc aagcattaac atgaacttaa attcatcaag gctaatctct
     1141 atatttgcct tgtgagtttt cttttgtgtt agttctttta ataaccactc ataaatcctc
     1201 atagagtatt tgttttcaaa agacttaaca tgttccagat tatattttat gaattttttt
     1261 aactggaaaa gataaggcaa tatctcttca ctaaaaacta attctaattt ttcgcttgag
     1321 aacttggcat agtttgtcca ctggaaaatc tcaaagcctt taaccaaagg attcctgatt
     1381 tccacagttc tcgtcatcag ctctctggtt gctttagcta atacaccata agcattttcc
     1441 ctactgatgt tcatcatctg agcgtattgg ttataagtga acgataccgt ccgttctttc
     1501 cttgtagggt tttcaatcgt ggggttgagt agtgccacac agcataaaat tagcttggtt
     1561 tcatgctccg ttaagtcata gcgactaatc gctagttcat ttgctttgaa aacaactaat
     1621 tcagacatac atctcaattg gtctaggtga ttttaatcac tataccaatt gagatgggct
     1681 agtcaatgat aattactagt ccttttcccg ggtgatctgg gtatctgtaa attctgctag
     1741 acctttgctg gaaaacttgt aaattctgct agaccctctg taaattccgc tagacctttg
     1801 tgtgtttttt ttgtttatat tcaagtggtt ataatttata gaataaagaa agaataaaaa
     1861 aagataaaaa gaatagatcc cagccctgtg tataactcac tactttagtc agttccgcag
     1921 tattacaaaa ggatgtcgca aacgctgttt gctcctctac aaaacagacc ttaaaaccct
     1981 aaaggcttaa gtagcaccct cgcaagctcg ggcaaatcgc tgaatattcc ttttgtctcc
     2041 gaccatcagg cacctgagtc gctgtctttt tcgtgacatt cagttcgctg cgctcacggc
     2101 tctggcagtg aatgggggta aatggcacta caggcgcctt ttatggattc atgcaaggaa
     2161 actacccata atacaagaaa agcccgtcac gggcttctca gggcgtttta tggcgggtct
     2221 gctatgtggt gctatctgac tttttgctgt tcagcagttc ctgccctctg attttccagt
     2281 ctgaccactt cggattatcc cgtgacaggt cattcagact ggctaatgca cccagtaagg
     2341 cagcggtatc atcaacaggc ttacccgtct tactgtccct agtgcttgga ttctcaccaa
     2401 taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca
     2461 ttactggatc tatcaacagg agtccaagcg agctctcgaa ccccagagtc ccgctcagaa
     2521 gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta
     2581 aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc
     2641 caacgctatg tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga
     2701 aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag
     2761 atcctcgccg tcgggcatgc gcgccttgag cctggcgaac agttcggctg gcgcgagccc
     2821 ctgatgctct tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc
     2881 tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg
     2941 cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga
     3001 caggagatcc tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac
     3061 aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc
     3121 ctcgtcctgc agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg
     3181 cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca
     3241 gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg
     3301 ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatca gatcatgatc ccctgcgcca
     3361 tcagatcctt ggcggcaaga aagccatcca gtttactttg cagggcttcc caaccttacc
     3421 agagggcgcc ccagctggca attccgacgt cggtgcctaa tgagtgagct aacttacatt
     3481 aattgcgttg cgcgctgatg tggtggagat cgaatttcaa aggttagttt tttcatgact
     3541 tccctctccc ccaaataaaa aggcctgctg gcgattacca gccagcaggc ctagtattaa
     3601 ccgtgtcctt ctcgatttga ataaataaaa acagccgttg ccagaaagag gcacggctgt
     3661 ttttattttc tattatttgt acagctcatc catgccaccg gtagaatgac gaccctccgc
     3721 gcgctcatat tgctctacga tcgtataatc ttcattatga gaggtgatgt ccagtttaat
     3781 attcacattg tacgcgccag gcagctgcac aggtttcttg gctttgtacg tggttttcac
     3841 ttcagcgtca taatggccgc catctttcag tttcaggcgc tgtttaattt cgcctttcag
     3901 cgcaccatct tccggataca tgcgttcgct agacgcctcc caacccatcg tctttttctg
     3961 catcaccggg ccatcagatg gaaaattagt accacgcagt ttaactttat agatgaactc
     4021 gccatcttgc agggaggagt cctgagtgac ggtcacgaca ccaccatctt caaaattcat
     4081 tacgcgttcc cacttgaaac cttccggaaa agacagcttc agatagtccg ggatatccgc
     4141 tgggtgttta acatacgctt tagaaccgta cataaattgc gggctcagga tgtcccacgc
     4201 aaaaggcagc gggccgcctt tagtcacttt cagtttggcg gtctgggtgc cttcatacgg
     4261 acggccctcg ccttcgcctt cgatctcgaa ctcgtgaccg ttaacagaac cctccatgtg
     4321 aactttgaag cgcatgaact ctttaatgat agccatgtta tcctcctcgc ccttggatct
     4381 tttgctagtc atgcggtacc tttctcctct ttaaagttaa agatcttcgt gctatggtac
     4441 atacattcac aaatgtatgt aaatctaacg cctgtaaatt cacgaacata tggcacgaaa
     4501 aaccaaacaa gaagcgcaag aaacgcgcca acacatcctc gatgtggctc tacgtctttt
     4561 ctcacagcag ggggtatcat ccacctcgct gggcgagatt gcaaaagcag ctggcgttac
     4621 gcgcggtgca atctactggc gtcgacgatc cggcaaacaa accaccgttg gtagcggtgg
     4681 tttttttgtt tggatcgaca atcttcgtaa gcgtcatcaa taagcgtaaa aaaaccgggc
     4741 aatgcccggt tttttaatga gaaattttac ctgtcgtagc cgccaccatc cggcaaagaa
     4801 gcatacaagg cttttggctt atagctacgt agcgcattgc gtcgcagcac aatcccggca
     4861 ccgatcgagt aatagtcaaa agcctccggt cggaggcttt tgactttctg cccggggcgt
     4921 acttgcttca taagcatcac gcaaaagctg caaaacagca tctttcccgg aaccagcatc
     4981 aagaactcgc cgttcgcttc ttcccctgaa atgattaact ccggtatcat gtggatcctt
     5041 aactttaaag aggagaaagg taccgcatgc gtaaaggaga agaacttttc actggagttg
     5101 tcccaattct tgttgaatta gatggtgatg ttaatgggca caaattttct gtcagtggag
     5161 agggtgaagg tgatgcaaca tacggaaaac ttacccttaa atttatttgc actactggaa
     5221 aactacctgt tccatggcca acacttgtca ctactttcgg ttatggtcta aaatgctttg
     5281 ctagataccc agatcatatg aaacggcatg actttttcaa gagtgccatg cccgaaggtt
     5341 atgtacagga aagaactata tttttcaaag atgacgggaa ctacaagaca cgtgctgaag
     5401 tcaagtttga aggtgatacc cttgttaata gaatcgagtt aaaaggtatt gattttaaag
     5461 aagatggaaa cattcttgga cacaaattgg aatacaacta taactcacac aatgtataca
     5521 tcatggcaga caaacaaaag aatggaatca aagttaactt caaaattaga cacaacattg
     5581 aagatggaag cgttcaacta gcagaccatt atcaacaaaa tactccaatt ggcgatggcc
     5641 ctgtcctttt accagacaac cattacctgt cctatcaatc tgccctttcg aaagatccca
     5701 acgaaaagag agaccacatg gtccttcttg agtttgtaac agctgctggg attacacatg
     5761 gcatggatga actatacaaa taataggctt aattagctga gtctagaggc atcaaataaa
     5821 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     5881 tctcctgagt aggacaaatc cgccgcccta ga
//
