LOCUS       sequence-244041-        5763 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  plasmid reporting the expression from constitutive and mutS 
            promoters.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5763)
  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 5763)
  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: Pconst-rfp + PmutS-yfp
FEATURES             Location/Qualifiers
     source          1..5763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1035..1046
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      complement(1359..1390)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     RBS             1532..1543
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1552..2265
                     /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(1582..1610)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2205..2225
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      2298..2384
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2430..2501
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2517..2544
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3075..4025)
                     /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(4787..4881)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4912..5706)
                     /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(5023..5042)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5633..5652
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 ccttggcggc aagaaagcca tccagtttac tttgcagggc ttcccaacct taccagaggg
       61 cgccccagct ggcaattccg acgtcggtgc ctaatgagtg agctaactta cattaattgc
      121 gttgcgcgct gatgtggtgg agatcgaatt tcaaaggtta gttttttcat gacttccctc
      181 tcccccaaat aaaaaggcct gctggcgatt accagccagc aggcctagta ttaaccgtgt
      241 ccttctcgat ttgaataaat aaaaacagcc gttgccagaa agaggcacgg ctgtttttat
      301 tttctattat ttgtacagct catccatgcc accggtagaa tgacgaccct ccgcgcgctc
      361 atattgctct acgatcgtat aatcttcatt atgagaggtg atgtccagtt taatattcac
      421 attgtacgcg ccaggcagct gcacaggttt cttggctttg tacgtggttt tcacttcagc
      481 gtcataatgg ccgccatctt tcagtttcag gcgctgttta atttcgcctt tcagcgcacc
      541 atcttccgga tacatgcgtt cgctagacgc ctcccaaccc atcgtctttt tctgcatcac
      601 cgggccatca gatggaaaat tagtaccacg cagtttaact ttatagatga actcgccatc
      661 ttgcagggag gagtcctgag tgacggtcac gacaccacca tcttcaaaat tcattacgcg
      721 ttcccacttg aaaccttccg gaaaagacag cttcagatag tccgggatat ccgctgggtg
      781 tttaacatac gctttagaac cgtacataaa ttgcgggctc aggatgtccc acgcaaaagg
      841 cagcgggccg cctttagtca ctttcagttt ggcggtctgg gtgccttcat acggacggcc
      901 ctcgccttcg ccttcgatct cgaactcgtg accgttaaca gaaccctcca tgtgaacttt
      961 gaagcgcatg aactctttaa tgatagccat gttatcctcc tcgcccttgg atcttttgct
     1021 agtcatgcgg tacctttctc ctctttaaag ttaaagatct gtgtccctct cgatggctgt
     1081 aatactccta taggttagac tttatgtcaa tactcttttt gataagtcga cgatccggca
     1141 aacaaaccac cgttggtagc ggtggttttt ttgtttggat cgacaatctt cgtaagcgtc
     1201 atcaataagc gtaaaaaaac cgggcaatgc ccggtttttt aatgagaaat tttacctgtc
     1261 gtagccgcca ccatccggca aagaagcata caaggctttt ggcttatagc tacgtagcgc
     1321 attgcgtcgc agcacaatcc cggcaccgat cgagtaatag tcaaaagcct ccggtcggag
     1381 gcttttgact ttctgcccgg ggcgtacttg cttcataagc atcacgcaaa agctgcaaaa
     1441 cagcatcttt cccggaacca gcatcaagaa ctcgccgttc gcttcttccc ctgaaatgat
     1501 taactccggt atcatgtgga tccttaactt taaagaggag aaaggtaccg catgcgtaaa
     1561 ggagaagaac ttttcactgg agttgtccca attcttgttg aattagatgg tgatgttaat
     1621 gggcacaaat tttctgtcag tggagagggt gaaggtgatg caacatacgg aaaacttacc
     1681 cttaaattta tttgcactac tggaaaacta cctgttccat ggccaacact tgtcactact
     1741 ttcggttatg gtctaaaatg ctttgctaga tacccagatc atatgaaacg gcatgacttt
     1801 ttcaagagtg ccatgcccga aggttatgta caggaaagaa ctatattttt caaagatgac
     1861 gggaactaca agacacgtgc tgaagtcaag tttgaaggtg atacccttgt taatagaatc
     1921 gagttaaaag gtattgattt taaagaagat ggaaacattc ttggacacaa attggaatac
     1981 aactataact cacacaatgt atacatcatg gcagacaaac aaaagaatgg aatcaaagtt
     2041 aacttcaaaa ttagacacaa cattgaagat ggaagcgttc aactagcaga ccattatcaa
     2101 caaaatactc caattggcga tggccctgtc cttttaccag acaaccatta cctgtcctat
     2161 caatctgccc tttcgaaaga tcccaacgaa aagagagacc acatggtcct tcttgagttt
     2221 gtaacagctg ctgggattac acatggcatg gatgaactat acaaataata ggcttaatta
     2281 gctgagtcta gaggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     2341 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ccctagagga
     2401 tccaaactcg agtaaggatc tccaggcatc aaataaaacg aaaggctcag tcgaaagact
     2461 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct
     2521 caccttcggg tgggcctttc tgcgtttata cctagggtac gggttttgct gcccgcaaac
     2581 gggctgttct ggtgttgcta gtttgttatc agaatcgcag atccggcttc agccggtttg
     2641 ccggctgaaa gcgctatttc ttccagaatt gccatgattt tttccccacg ggaggcgtca
     2701 ctggctcccg tgttgtcggc agctttgatt cgataagcag catcgcctgt ttcaggctgt
     2761 ctatgtgtga ctgttgagct gtaacaagtt gtctcaggtg ttcaatttca tgttctagtt
     2821 gctttgtttt actggtttca cctgttctat taggtgttac atgctgttca tctgttacat
     2881 tgtcgatctg ttcatggtga acagctttga atgcaccaaa aactcgtaaa agctctgatg
     2941 tatctatctt ttttacaccg ttttcatctg tgcatatgga cagttttccc tttgatatgt
     3001 aacggtgaac agttgttcta cttttgtttg ttagtcttga tgcttcactg atagatacaa
     3061 gagccataag aacctcagat ccttccgtat ttagccagta tgttctctag tgtggttcgt
     3121 tgtttttgcg tgagccatga gaacgaacca ttgagatcat acttactttg catgtcactc
     3181 aaaaattttg cctcaaaact ggtgagctga atttttgcag ttaaagcatc gtgtagtgtt
     3241 tttcttagtc cgttatgtag gtaggaatct gatgtaatgg ttgttggtat tttgtcacca
     3301 ttcattttta tctggttgtt ctcaagttcg gttacgagat ccatttgtct atctagttca
     3361 acttggaaaa tcaacgtatc agtcgggcgg cctcgcttat caaccaccaa tttcatattg
     3421 ctgtaagtgt ttaaatcttt acttattggt ttcaaaaccc attggttaag ccttttaaac
     3481 tcatggtagt tattttcaag cattaacatg aacttaaatt catcaaggct aatctctata
     3541 tttgccttgt gagttttctt ttgtgttagt tcttttaata accactcata aatcctcata
     3601 gagtatttgt tttcaaaaga cttaacatgt tccagattat attttatgaa tttttttaac
     3661 tggaaaagat aaggcaatat ctcttcacta aaaactaatt ctaatttttc gcttgagaac
     3721 ttggcatagt ttgtccactg gaaaatctca aagcctttaa ccaaaggatt cctgatttcc
     3781 acagttctcg tcatcagctc tctggttgct ttagctaata caccataagc attttcccta
     3841 ctgatgttca tcatctgagc gtattggtta taagtgaacg ataccgtccg ttctttcctt
     3901 gtagggtttt caatcgtggg gttgagtagt gccacacagc ataaaattag cttggtttca
     3961 tgctccgtta agtcatagcg actaatcgct agttcatttg ctttgaaaac aactaattca
     4021 gacatacatc tcaattggtc taggtgattt taatcactat accaattgag atgggctagt
     4081 caatgataat tactagtcct tttcccgggt gatctgggta tctgtaaatt ctgctagacc
     4141 tttgctggaa aacttgtaaa ttctgctaga ccctctgtaa attccgctag acctttgtgt
     4201 gttttttttg tttatattca agtggttata atttatagaa taaagaaaga ataaaaaaag
     4261 ataaaaagaa tagatcccag ccctgtgtat aactcactac tttagtcagt tccgcagtat
     4321 tacaaaagga tgtcgcaaac gctgtttgct cctctacaaa acagacctta aaaccctaaa
     4381 ggcttaagta gcaccctcgc aagctcgggc aaatcgctga atattccttt tgtctccgac
     4441 catcaggcac ctgagtcgct gtctttttcg tgacattcag ttcgctgcgc tcacggctct
     4501 ggcagtgaat gggggtaaat ggcactacag gcgcctttta tggattcatg caaggaaact
     4561 acccataata caagaaaagc ccgtcacggg cttctcaggg cgttttatgg cgggtctgct
     4621 atgtggtgct atctgacttt ttgctgttca gcagttcctg ccctctgatt ttccagtctg
     4681 accacttcgg attatcccgt gacaggtcat tcagactggc taatgcaccc agtaaggcag
     4741 cggtatcatc aacaggctta cccgtcttac tgtccctagt gcttggattc tcaccaataa
     4801 aaaacgcccg gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta
     4861 ctggatctat caacaggagt ccaagcgagc tctcgaaccc cagagtcccg ctcagaagaa
     4921 ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag
     4981 cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa
     5041 cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa
     5101 gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc
     5161 ctcgccgtcg ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg
     5221 atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg
     5281 ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag
     5341 ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag
     5401 gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac
     5461 gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc
     5521 gtcctgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc
     5581 ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc
     5641 atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc
     5701 aatcatgcga aacgatcctc atcctgtctc ttgatcagat catgatcccc tgcgccatca
     5761 gat
//
