LOCUS       sequence-454158-        5994 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive promoter driving expression of CFP reporter and gadX 
            promoter driving expression of YFP reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5994)
  AUTHORS   Alnahhas RN, Andreani V, Dunlop MJ
  TITLE     Evaluating the predictive power of combined gene expression dynamics
            from single cells on antibiotic survival.
  JOURNAL   mSystems. 2025 May 20:e0158824. doi: 10.1128/msystems.01588-24.
  PUBMED    40391890
REFERENCE   2  (bases 1 to 5994)
  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: "mSystems. 
            2025 May 20:e0158824. doi: 10.1128/msystems.01588-24."
COMMENT     Sequence Label: Pconst-CFP--PgadX-YFP
FEATURES             Location/Qualifiers
     source          1..5994
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      701..923
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(1415..1509)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1540..2334)
                     /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(1651..1670)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2261..2280
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2513..2531
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     2654..2673
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     terminator      3617..3703
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(3891..3962)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(4003..4716)
                     /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="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             complement(join(5697..5994,1..653))
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
ORIGIN
        1 aacgtatcag tcgggcggcc tcgcttatca accaccaatt tcatattgct gtaagtgttt
       61 aaatctttac ttattggttt caaaacccat tggttaagcc ttttaaactc atggtagtta
      121 ttttcaagca ttaacatgaa cttaaattca tcaaggctaa tctctatatt tgccttgtga
      181 gttttctttt gtgttagttc ttttaataac cactcataaa tcctcataga gtatttgttt
      241 tcaaaagact taacatgttc cagattatat tttatgaatt tttttaactg gaaaagataa
      301 ggcaatatct cttcactaaa aactaattct aatttttcgc ttgagaactt ggcatagttt
      361 gtccactgga aaatctcaaa gcctttaacc aaaggattcc tgatttccac agttctcgtc
      421 atcagctctc tggttgcttt agctaataca ccataagcat tttccctact gatgttcatc
      481 atctgagcgt attggttata agtgaacgat accgtccgtt ctttccttgt agggttttca
      541 atcgtggggt tgagtagtgc cacacagcat aaaattagct tggtttcatg ctccgttaag
      601 tcatagcgac taatcgctag ttcatttgct ttgaaaacaa ctaattcaga catacatctc
      661 aattggtcta ggtgatttta atcactatac caattgagat gggctagtca atgataatta
      721 ctagtccttt tcctttgagt tgtgggtatc tgtaaattct gctagacctt tgctggaaaa
      781 cttgtaaatt ctgctagacc ctctgtaaat tccgctagac ctttgtgtgt tttttttgtt
      841 tatattcaag tggttataat ttatagaata aagaaagaat aaaaaaagat aaaaagaata
      901 gatcccagcc ctgtgtataa ctcactactt tagtcagttc cgcagtatta caaaaggatg
      961 tcgcaaacgc tgtttgctcc tctacaaaac agaccttaaa accctaaagg cttaagtagc
     1021 accctcgcaa gctcgggcaa atcgctgaat attccttttg tctccgacca tcaggcacct
     1081 gagtcgctgt ctttttcgtg acattcagtt cgctgcgctc acggctctgg cagtgaatgg
     1141 gggtaaatgg cactacaggc gccttttatg gattcatgca aggaaactac ccataataca
     1201 agaaaagccc gtcacgggct tctcagggcg ttttatggcg ggtctgctat gtggtgctat
     1261 ctgacttttt gctgttcagc agttcctgcc ctctgatttt ccagtctgac cacttcggat
     1321 tatcccgtga caggtcattc agactggcta atgcacccag taaggcagcg gtatcatcaa
     1381 caggcttacc cgtcttactg tccctagtgc ttggattctc accaataaaa aacgcccggc
     1441 ggcaaccgag cgttctgaac aaatccagat ggagttctga ggtcattact ggatctatca
     1501 acaggagtcc aagcgagctc tcgaacccca gagtcccgct cagaagaact cgtcaagaag
     1561 gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg
     1621 gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg
     1681 atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc
     1741 caccatgata ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg
     1801 catgcgcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc
     1861 cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg
     1921 tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc
     1981 atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc
     2041 cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc
     2101 tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cctgcagttc
     2161 attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag
     2221 ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag
     2281 cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa
     2341 cgatcctcat cctgtctctt gatcagatct tgatcccctg cgccatcaga tccttggcgg
     2401 caagaaagcc atccagttta ctttgcaggg cttcccaacc ttaccagagg gcgccccagc
     2461 tggcaattcc gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     2521 tatcacgagg ccctttcgtc ttcgcatgca aggagatggc gcccaacagt cccccggcca
     2581 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
     2641 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     2701 tgatgccggc cacgatgcgt ccggcgtaga ggatcgagtt atcaaaaaga gtattgtctt
     2761 aaagtctaac ctataggaaa attacagcca tcgagaggga cacggcgaaa cctcgagggg
     2821 atcctctaga tttaagaagg agatatacat atgactagca aaagaagcaa aggtgaagaa
     2881 ctgttcactg gtgttgttcc aattctggtt gaactggatg gtgatgttaa tggtcacaaa
     2941 ttttctgtct ctggtgaggg tgaaggtgat gcaacctacg gtaaactgac cctgaaattt
     3001 atttgcacta ctggtaaact gcctgttccg tggccaaccc tggtcactac tctgacttgg
     3061 ggtgttcaat gctttgctcg ttacccagat cacatgaaac agcatgactt tttcaagtct
     3121 gccatgccgg aaggttatgt tcaggaacgt actatctttt tcaaagatga cggtaactac
     3181 aagacccgtg ctgaagtcaa gtttgaaggt gataccctgg ttaatcgtat cgagctgaaa
     3241 ggtattgatt ttaaagaaga tggtaacatt ctgggtcaca aactggaata caacgctatt
     3301 tctgataatg tatacatcac tgctgacaaa caaaagaatg gtatcaaagc taatttcaaa
     3361 attcgtcaca acattgaaga tggtagcgtt caactggcag accattatca acaaaatact
     3421 ccaattggcg atggccctgt cctgctgcca gacaaccatt acctgtccac ccaatctcgt
     3481 ctgtctaaag atccgaacga aaagcgcgat cacatggtcc tgctggagtt tgtaaccgct
     3541 gctggtatta ccctgggcat ggatgaactg tataaataaa tgtccagacc tgcaggcatg
     3601 caagctctag aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     3661 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgc cctagaccta
     3721 gggtacgggt tttgctgccc gcaaacgggc tgttctggtg ttgctagttt gttatcagaa
     3781 tcgcagatcc ggcttcagcc ggtttgccgg ctgaaagcgc tatttcttcc agaattgcca
     3841 tgattttttc cccacgggag gcgtcactgg ctcccgtgtt gtcggcagct gagagcgttc
     3901 accgacaaac aacagataaa acgaaaggcc cagtctttcg actgagcctt tcgttttatt
     3961 tgatgcctct agagcttgca tgcctgcagg tctggacatt tatttgtata gttcatccat
     4021 gccatgtgta atcccagcag ctgttacaaa ctcaagaagg accatgtggt ctctcttttc
     4081 gttgggatct ttcgaaaggg cagattgata ggacaggtaa tggttgtctg gtaaaaggac
     4141 agggccatcg ccaattggag tattttgttg ataatggtct gctagttgaa cgcttccatc
     4201 ttcaatgttg tgtctaattt tgaagttaac tttgattcca ttcttttgtt tgtctgccat
     4261 gatgtataca ttgtgtgagt tatagttgta ttccaatttg tgtccaagaa tgtttccatc
     4321 ttctttaaaa tcaatacctt ttaactcgat tctattaaca agggtatcac cttcaaactt
     4381 gacttcagca cgtgtcttgt agttcccgtc atctttgaaa aatatagttc tttcctgtac
     4441 ataaccttcg ggcatggcac tcttgaaaaa gtcatgctgt ttcatatgat ctgggtatct
     4501 cgcaaagcat tgaagaccat aaccgaaagt agtgacaagt gttggccatg gaacaggtag
     4561 ttttccagta gtgcaaataa atttaagggt aagttttccg tatgttgcat caccttcacc
     4621 ctctccactg acagaaaatt tgtgcccatt aacatcacca tctaattcaa caagaattgg
     4681 gacaactcca gtgaaaagtt cttctccttt actcatatgt atatctcctt cttaaatcta
     4741 gaggatctct cgagaggtca ccgccattaa aatagcgata ttcaccatta accatggtga
     4801 gaatatattt atgtcttgca tacgcaatta gacaattccc atgtagtgat tgcatagttg
     4861 acttaatatt acataaacat attactgttt attaatgtag cacgccgccc tcttaagtca
     4921 aattgattga taaataaatt taaacgaagc gggagaacag gatgataatg tgtgatgttc
     4981 tacgggcagg atgactggat ttataataca aaaacgtgtt taagaacaca gggaaattaa
     5041 tagtaagcaa ataatccctt ttcgtgacat taaaggtaat cgctacattt aataaacatt
     5101 catataacat atatcttatc aacacgatga atagacagcc aatatattat tgcgattaat
     5161 aagcaaccga atgcccagct gtttttttaa aggctgggca ttcggttttt acaacgttat
     5221 gttatcaggt gtgtttaaag ctgttctgct gggcaatacc ctgcagtttc gggtgatcgc
     5281 tgagatattt cagggaggct ttgtagtctt ccagcaggat ccatcgaggt gaagacgaaa
     5341 gggcctttga ttcgataagc agcatcgcct gtttcaggct gtctatgtgt gactgttgag
     5401 ctgtaacaag ttgtctcagg tgttcaattt catgttctag ttgctttgtt ttactggttt
     5461 cacctgttct attaggtgtt acatgctgtt catctgttac attgtcgatc tgttcatggt
     5521 gaacagcttt gaatgcacca aaaactcgta aaagctctga tgtatctatc ttttttacac
     5581 cgttttcatc tgtgcatatg gacagttttc cctttgatat gtaacggtga acagttgttc
     5641 tacttttgtt tgttagtctt gatgcttcac tgatagatac aagagccata agaacctcag
     5701 atccttccgt atttagccag tatgttctct agtgtggttc gttgtttttg cgtgagccat
     5761 gagaacgaac cattgagatc atacttactt tgcatgtcac tcaaaaattt tgcctcaaaa
     5821 ctggtgagct gaatttttgc agttaaagca tcgtgtagtg tttttcttag tccgttatgt
     5881 aggtaggaat ctgatgtaat ggttgttggt attttgtcac cattcatttt tatctggttg
     5941 ttctcaagtt cggttacgag atccatttgt ctatctagtt caacttggaa aatc
//
