LOCUS       sequence-238420-        5861 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  plasmid reporting the expression from acrAB and constitutive 
            promoters.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5861)
  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 5861)
  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 + Pconst-yfp
FEATURES             Location/Qualifiers
     source          1..5861
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             15..26
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             35..748
                     /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(65..93)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     688..708
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      781..867
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      913..984
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1000..1027
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1558..2508)
                     /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(3270..3364)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3395..4189)
                     /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(3506..3525)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4116..4135
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     RBS             5281..5292
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      complement(5754..5785)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 ggatccttaa ctttaaagag gagaaaggta ccgcatgcgt aaaggagaag aacttttcac
       61 tggagttgtc ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt
      121 cagtggagag ggtgaaggtg atgcaacata cggaaaactt acccttaaat ttatttgcac
      181 tactggaaaa ctacctgttc catggccaac acttgtcact actttcggtt atggtctaaa
      241 atgctttgct agatacccag atcatatgaa acggcatgac tttttcaaga gtgccatgcc
      301 cgaaggttat gtacaggaaa gaactatatt tttcaaagat gacgggaact acaagacacg
      361 tgctgaagtc aagtttgaag gtgataccct tgttaataga atcgagttaa aaggtattga
      421 ttttaaagaa gatggaaaca ttcttggaca caaattggaa tacaactata actcacacaa
      481 tgtatacatc atggcagaca aacaaaagaa tggaatcaaa gttaacttca aaattagaca
      541 caacattgaa gatggaagcg ttcaactagc agaccattat caacaaaata ctccaattgg
      601 cgatggccct gtccttttac cagacaacca ttacctgtcc tatcaatctg ccctttcgaa
      661 agatcccaac gaaaagagag accacatggt ccttcttgag tttgtaacag ctgctgggat
      721 tacacatggc atggatgaac tatacaaata ataggcttaa ttagctgagt ctagaggcat
      781 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
      841 gtgaacgctc tcctgagtag gacaaatccg ccgccctaga ggatccaaac tcgagtaagg
      901 atctccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
      961 tgttgtttgt cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct
     1021 ttctgcgttt atacctaggg tacgggtttt gctgcccgca aacgggctgt tctggtgttg
     1081 ctagtttgtt atcagaatcg cagatccggc ttcagccggt ttgccggctg aaagcgctat
     1141 ttcttccaga attgccatga ttttttcccc acgggaggcg tcactggctc ccgtgttgtc
     1201 ggcagctttg attcgataag cagcatcgcc tgtttcaggc tgtctatgtg tgactgttga
     1261 gctgtaacaa gttgtctcag gtgttcaatt tcatgttcta gttgctttgt tttactggtt
     1321 tcacctgttc tattaggtgt tacatgctgt tcatctgtta cattgtcgat ctgttcatgg
     1381 tgaacagctt tgaatgcacc aaaaactcgt aaaagctctg atgtatctat cttttttaca
     1441 ccgttttcat ctgtgcatat ggacagtttt ccctttgata tgtaacggtg aacagttgtt
     1501 ctacttttgt ttgttagtct tgatgcttca ctgatagata caagagccat aagaacctca
     1561 gatccttccg tatttagcca gtatgttctc tagtgtggtt cgttgttttt gcgtgagcca
     1621 tgagaacgaa ccattgagat catacttact ttgcatgtca ctcaaaaatt ttgcctcaaa
     1681 actggtgagc tgaatttttg cagttaaagc atcgtgtagt gtttttctta gtccgttatg
     1741 taggtaggaa tctgatgtaa tggttgttgg tattttgtca ccattcattt ttatctggtt
     1801 gttctcaagt tcggttacga gatccatttg tctatctagt tcaacttgga aaatcaacgt
     1861 atcagtcggg cggcctcgct tatcaaccac caatttcata ttgctgtaag tgtttaaatc
     1921 tttacttatt ggtttcaaaa cccattggtt aagcctttta aactcatggt agttattttc
     1981 aagcattaac atgaacttaa attcatcaag gctaatctct atatttgcct tgtgagtttt
     2041 cttttgtgtt agttctttta ataaccactc ataaatcctc atagagtatt tgttttcaaa
     2101 agacttaaca tgttccagat tatattttat gaattttttt aactggaaaa gataaggcaa
     2161 tatctcttca ctaaaaacta attctaattt ttcgcttgag aacttggcat agtttgtcca
     2221 ctggaaaatc tcaaagcctt taaccaaagg attcctgatt tccacagttc tcgtcatcag
     2281 ctctctggtt gctttagcta atacaccata agcattttcc ctactgatgt tcatcatctg
     2341 agcgtattgg ttataagtga acgataccgt ccgttctttc cttgtagggt tttcaatcgt
     2401 ggggttgagt agtgccacac agcataaaat tagcttggtt tcatgctccg ttaagtcata
     2461 gcgactaatc gctagttcat ttgctttgaa aacaactaat tcagacatac atctcaattg
     2521 gtctaggtga ttttaatcac tataccaatt gagatgggct agtcaatgat aattactagt
     2581 ccttttcccg ggtgatctgg gtatctgtaa attctgctag acctttgctg gaaaacttgt
     2641 aaattctgct agaccctctg taaattccgc tagacctttg tgtgtttttt ttgtttatat
     2701 tcaagtggtt ataatttata gaataaagaa agaataaaaa aagataaaaa gaatagatcc
     2761 cagccctgtg tataactcac tactttagtc agttccgcag tattacaaaa ggatgtcgca
     2821 aacgctgttt gctcctctac aaaacagacc ttaaaaccct aaaggcttaa gtagcaccct
     2881 cgcaagctcg ggcaaatcgc tgaatattcc ttttgtctcc gaccatcagg cacctgagtc
     2941 gctgtctttt tcgtgacatt cagttcgctg cgctcacggc tctggcagtg aatgggggta
     3001 aatggcacta caggcgcctt ttatggattc atgcaaggaa actacccata atacaagaaa
     3061 agcccgtcac gggcttctca gggcgtttta tggcgggtct gctatgtggt gctatctgac
     3121 tttttgctgt tcagcagttc ctgccctctg attttccagt ctgaccactt cggattatcc
     3181 cgtgacaggt cattcagact ggctaatgca cccagtaagg cagcggtatc atcaacaggc
     3241 ttacccgtct tactgtccct agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa
     3301 ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg
     3361 agtccaagcg agctctcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat
     3421 agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag
     3481 cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc
     3541 ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca
     3601 tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc
     3661 gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat
     3721 catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg
     3781 cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag
     3841 ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca
     3901 cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc
     3961 aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca
     4021 gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga
     4081 acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct
     4141 ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc
     4201 ctcatcctgt ctcttgatca gatcatgatc ccctgcgcca tcagatcctt ggcggcaaga
     4261 aagccatcca gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca
     4321 attccgacgt cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgcgctgatg
     4381 tggtggagat cgaatttcaa aggttagttt tttcatgact tccctctccc ccaaataaaa
     4441 aggcctgctg gcgattacca gccagcaggc ctagtattaa ccgtgtcctt ctcgatttga
     4501 ataaataaaa acagccgttg ccagaaagag gcacggctgt ttttattttc tattatttgt
     4561 acagctcatc catgccaccg gtagaatgac gaccctccgc gcgctcatat tgctctacga
     4621 tcgtataatc ttcattatga gaggtgatgt ccagtttaat attcacattg tacgcgccag
     4681 gcagctgcac aggtttcttg gctttgtacg tggttttcac ttcagcgtca taatggccgc
     4741 catctttcag tttcaggcgc tgtttaattt cgcctttcag cgcaccatct tccggataca
     4801 tgcgttcgct agacgcctcc caacccatcg tctttttctg catcaccggg ccatcagatg
     4861 gaaaattagt accacgcagt ttaactttat agatgaactc gccatcttgc agggaggagt
     4921 cctgagtgac ggtcacgaca ccaccatctt caaaattcat tacgcgttcc cacttgaaac
     4981 cttccggaaa agacagcttc agatagtccg ggatatccgc tgggtgttta acatacgctt
     5041 tagaaccgta cataaattgc gggctcagga tgtcccacgc aaaaggcagc gggccgcctt
     5101 tagtcacttt cagtttggcg gtctgggtgc cttcatacgg acggccctcg ccttcgcctt
     5161 cgatctcgaa ctcgtgaccg ttaacagaac cctccatgtg aactttgaag cgcatgaact
     5221 ctttaatgat agccatgtta tcctcctcgc ccttggatct tttgctagtc atgcggtacc
     5281 tttctcctct ttaaagttaa agatcttcgt gctatggtac atacattcac aaatgtatgt
     5341 aaatctaacg cctgtaaatt cacgaacata tggcacgaaa aaccaaacaa gaagcgcaag
     5401 aaacgcgcca acacatcctc gatgtggctc tacgtctttt ctcacagcag ggggtatcat
     5461 ccacctcgct gggcgagatt gcaaaagcag ctggcgttac gcgcggtgca atctactggc
     5521 gtcgacgatc cggcaaacaa accaccgttg gtagcggtgg tttttttgtt tggatcgaca
     5581 atcttcgtaa gcgtcatcaa taagcgtaaa aaaaccgggc aatgcccggt tttttaatga
     5641 gaaattttac ctgtcgtagc cgccaccatc cggcaaagaa gcatacaagg cttttggctt
     5701 atagctacgt agcgcattgc gtcgcagcac aatcccggca ccgatcgagt aatagtcaaa
     5761 agcctccggt cggaggcttt tgactttctg cccgggttat caaaaagagt attgacataa
     5821 agtctaacct ataggagtat tacagccatc gagagggaca c
//
