LOCUS       sequence-231445-        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"
     terminator      complement(234..265)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     RBS             407..418
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             427..1140
                     /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(457..485)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1080..1100
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1173..1259
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1305..1376
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1392..1419
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1950..2900)
                     /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(3662..3756)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3787..4581)
                     /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(3898..3917)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4508..4527
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     RBS             5673..5684
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 gtcgacgatc cggcaaacaa accaccgttg gtagcggtgg tttttttgtt tggatcgaca
       61 atcttcgtaa gcgtcatcaa taagcgtaaa aaaaccgggc aatgcccggt tttttaatga
      121 gaaattttac ctgtcgtagc cgccaccatc cggcaaagaa gcatacaagg cttttggctt
      181 atagctacgt agcgcattgc gtcgcagcac aatcccggca ccgatcgagt aatagtcaaa
      241 agcctccggt cggaggcttt tgactttctg cccggggcgt acttgcttca taagcatcac
      301 gcaaaagctg caaaacagca tctttcccgg aaccagcatc aagaactcgc cgttcgcttc
      361 ttcccctgaa atgattaact ccggtatcat gtggatcctt aactttaaag aggagaaagg
      421 taccgcatgc gtaaaggaga agaacttttc actggagttg tcccaattct tgttgaatta
      481 gatggtgatg ttaatgggca caaattttct gtcagtggag agggtgaagg tgatgcaaca
      541 tacggaaaac ttacccttaa atttatttgc actactggaa aactacctgt tccatggcca
      601 acacttgtca ctactttcgg ttatggtcta aaatgctttg ctagataccc agatcatatg
      661 aaacggcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga aagaactata
      721 tttttcaaag atgacgggaa ctacaagaca cgtgctgaag tcaagtttga aggtgatacc
      781 cttgttaata gaatcgagtt aaaaggtatt gattttaaag aagatggaaa cattcttgga
      841 cacaaattgg aatacaacta taactcacac aatgtataca tcatggcaga caaacaaaag
      901 aatggaatca aagttaactt caaaattaga cacaacattg aagatggaag cgttcaacta
      961 gcagaccatt atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt accagacaac
     1021 cattacctgt cctatcaatc tgccctttcg aaagatccca acgaaaagag agaccacatg
     1081 gtccttcttg agtttgtaac agctgctggg attacacatg gcatggatga actatacaaa
     1141 taataggctt aattagctga gtctagaggc atcaaataaa acgaaaggct cagtcgaaag
     1201 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc
     1261 cgccgcccta gaggatccaa actcgagtaa ggatctccag gcatcaaata aaacgaaagg
     1321 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctctact
     1381 agagtcacac tggctcacct tcgggtgggc ctttctgcgt ttatacctag ggtacgggtt
     1441 ttgctgcccg caaacgggct gttctggtgt tgctagtttg ttatcagaat cgcagatccg
     1501 gcttcagccg gtttgccggc tgaaagcgct atttcttcca gaattgccat gattttttcc
     1561 ccacgggagg cgtcactggc tcccgtgttg tcggcagctt tgattcgata agcagcatcg
     1621 cctgtttcag gctgtctatg tgtgactgtt gagctgtaac aagttgtctc aggtgttcaa
     1681 tttcatgttc tagttgcttt gttttactgg tttcacctgt tctattaggt gttacatgct
     1741 gttcatctgt tacattgtcg atctgttcat ggtgaacagc tttgaatgca ccaaaaactc
     1801 gtaaaagctc tgatgtatct atctttttta caccgttttc atctgtgcat atggacagtt
     1861 ttccctttga tatgtaacgg tgaacagttg ttctactttt gtttgttagt cttgatgctt
     1921 cactgataga tacaagagcc ataagaacct cagatccttc cgtatttagc cagtatgttc
     1981 tctagtgtgg ttcgttgttt ttgcgtgagc catgagaacg aaccattgag atcatactta
     2041 ctttgcatgt cactcaaaaa ttttgcctca aaactggtga gctgaatttt tgcagttaaa
     2101 gcatcgtgta gtgtttttct tagtccgtta tgtaggtagg aatctgatgt aatggttgtt
     2161 ggtattttgt caccattcat ttttatctgg ttgttctcaa gttcggttac gagatccatt
     2221 tgtctatcta gttcaacttg gaaaatcaac gtatcagtcg ggcggcctcg cttatcaacc
     2281 accaatttca tattgctgta agtgtttaaa tctttactta ttggtttcaa aacccattgg
     2341 ttaagccttt taaactcatg gtagttattt tcaagcatta acatgaactt aaattcatca
     2401 aggctaatct ctatatttgc cttgtgagtt ttcttttgtg ttagttcttt taataaccac
     2461 tcataaatcc tcatagagta tttgttttca aaagacttaa catgttccag attatatttt
     2521 atgaattttt ttaactggaa aagataaggc aatatctctt cactaaaaac taattctaat
     2581 ttttcgcttg agaacttggc atagtttgtc cactggaaaa tctcaaagcc tttaaccaaa
     2641 ggattcctga tttccacagt tctcgtcatc agctctctgg ttgctttagc taatacacca
     2701 taagcatttt ccctactgat gttcatcatc tgagcgtatt ggttataagt gaacgatacc
     2761 gtccgttctt tccttgtagg gttttcaatc gtggggttga gtagtgccac acagcataaa
     2821 attagcttgg tttcatgctc cgttaagtca tagcgactaa tcgctagttc atttgctttg
     2881 aaaacaacta attcagacat acatctcaat tggtctaggt gattttaatc actataccaa
     2941 ttgagatggg ctagtcaatg ataattacta gtccttttcc cgggtgatct gggtatctgt
     3001 aaattctgct agacctttgc tggaaaactt gtaaattctg ctagaccctc tgtaaattcc
     3061 gctagacctt tgtgtgtttt ttttgtttat attcaagtgg ttataattta tagaataaag
     3121 aaagaataaa aaaagataaa aagaatagat cccagccctg tgtataactc actactttag
     3181 tcagttccgc agtattacaa aaggatgtcg caaacgctgt ttgctcctct acaaaacaga
     3241 ccttaaaacc ctaaaggctt aagtagcacc ctcgcaagct cgggcaaatc gctgaatatt
     3301 ccttttgtct ccgaccatca ggcacctgag tcgctgtctt tttcgtgaca ttcagttcgc
     3361 tgcgctcacg gctctggcag tgaatggggg taaatggcac tacaggcgcc ttttatggat
     3421 tcatgcaagg aaactaccca taatacaaga aaagcccgtc acgggcttct cagggcgttt
     3481 tatggcgggt ctgctatgtg gtgctatctg actttttgct gttcagcagt tcctgccctc
     3541 tgattttcca gtctgaccac ttcggattat cccgtgacag gtcattcaga ctggctaatg
     3601 cacccagtaa ggcagcggta tcatcaacag gcttacccgt cttactgtcc ctagtgcttg
     3661 gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga
     3721 gttctgaggt cattactgga tctatcaaca ggagtccaag cgagctctcg aaccccagag
     3781 tcccgctcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc
     3841 ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat
     3901 atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc
     3961 gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccatg
     4021 ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg agcctggcga acagttcggc
     4081 tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat
     4141 ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg
     4201 atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc
     4261 aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc
     4321 cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga
     4381 tagccgcgct gcctcgtcct gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa
     4441 aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt
     4501 ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg
     4561 caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat cagatcatga
     4621 tcccctgcgc catcagatcc ttggcggcaa gaaagccatc cagtttactt tgcagggctt
     4681 cccaacctta ccagagggcg ccccagctgg caattccgac gtcggtgcct aatgagtgag
     4741 ctaacttaca ttaattgcgt tgcgcgctga tgtggtggag atcgaatttc aaaggttagt
     4801 tttttcatga cttccctctc ccccaaataa aaaggcctgc tggcgattac cagccagcag
     4861 gcctagtatt aaccgtgtcc ttctcgattt gaataaataa aaacagccgt tgccagaaag
     4921 aggcacggct gtttttattt tctattattt gtacagctca tccatgccac cggtagaatg
     4981 acgaccctcc gcgcgctcat attgctctac gatcgtataa tcttcattat gagaggtgat
     5041 gtccagttta atattcacat tgtacgcgcc aggcagctgc acaggtttct tggctttgta
     5101 cgtggttttc acttcagcgt cataatggcc gccatctttc agtttcaggc gctgtttaat
     5161 ttcgcctttc agcgcaccat cttccggata catgcgttcg ctagacgcct cccaacccat
     5221 cgtctttttc tgcatcaccg ggccatcaga tggaaaatta gtaccacgca gtttaacttt
     5281 atagatgaac tcgccatctt gcagggagga gtcctgagtg acggtcacga caccaccatc
     5341 ttcaaaattc attacgcgtt cccacttgaa accttccgga aaagacagct tcagatagtc
     5401 cgggatatcc gctgggtgtt taacatacgc tttagaaccg tacataaatt gcgggctcag
     5461 gatgtcccac gcaaaaggca gcgggccgcc tttagtcact ttcagtttgg cggtctgggt
     5521 gccttcatac ggacggccct cgccttcgcc ttcgatctcg aactcgtgac cgttaacaga
     5581 accctccatg tgaactttga agcgcatgaa ctctttaatg atagccatgt tatcctcctc
     5641 gcccttggat cttttgctag tcatgcggta cctttctcct ctttaaagtt aaagatctgt
     5701 gtccctctcg atggctgtaa tactcctata ggttagactt tatgtcaata ctctttttga
     5761 taa
//
