LOCUS       sequence-454186-        5860 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive promoter driving expression of CFP reporter and recA 
            promoter driving expression of YFP reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5860)
  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 5860)
  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--PrecA-YFP
FEATURES             Location/Qualifiers
     source          1..5860
                     /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(4115..4186)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(4227..4940)
                     /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(5563..5860,1..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 accgaccggt cggcagctga gagcgttcac cgaccggtcg gcagctgaga gcgttcaccg
     3961 accggtcggc agctgagagc gttcaccgac cggtcggcag ctgagagcgt tcaccgaccg
     4021 gtcggcagct gagagcgttc accgaccggt cggcagctga gagcgttcac cgaccggtcg
     4081 gcagctgaga gcgttcaccg accggtcggc agctgagagc gttcaccgac aaacaacaga
     4141 taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctctagagct
     4201 tgcatgcctg caggtctgga catttatttg tatagttcat ccatgccatg tgtaatccca
     4261 gcagctgtta caaactcaag aaggaccatg tggtctctct tttcgttggg atctttcgaa
     4321 agggcagatt gataggacag gtaatggttg tctggtaaaa ggacagggcc atcgccaatt
     4381 ggagtatttt gttgataatg gtctgctagt tgaacgcttc catcttcaat gttgtgtcta
     4441 attttgaagt taactttgat tccattcttt tgtttgtctg ccatgatgta tacattgtgt
     4501 gagttatagt tgtattccaa tttgtgtcca agaatgtttc catcttcttt aaaatcaata
     4561 ccttttaact cgattctatt aacaagggta tcaccttcaa acttgacttc agcacgtgtc
     4621 ttgtagttcc cgtcatcttt gaaaaatata gttctttcct gtacataacc ttcgggcatg
     4681 gcactcttga aaaagtcatg ctgtttcata tgatctgggt atctcgcaaa gcattgaaga
     4741 ccataaccga aagtagtgac aagtgttggc catggaacag gtagttttcc agtagtgcaa
     4801 ataaatttaa gggtaagttt tccgtatgtt gcatcacctt caccctctcc actgacagaa
     4861 aatttgtgcc cattaacatc accatctaat tcaacaagaa ttgggacaac tccagtgaaa
     4921 agttcttctc ctttactcat atgtatatct ccttcttaaa tctagaggat ctctcgagcg
     4981 catgatggag cctttaccaa attgtttctc aatctggccc agtgctgccg ccaacgcttt
     5041 ctgtttgttt tcgtcgatag ccatttttac tcctgtcatg ccgggtaata ccggatagtc
     5101 aatatgttct gttgaagcaa ttatactgta tgctcataca gtatcaagtg ttttgtagaa
     5161 attgttgcca caaggtctgc aaggatccat cgaggtgaag acgaaagggc ctttgattcg
     5221 ataagcagca tcgcctgttt caggctgtct atgtgtgact gttgagctgt aacaagttgt
     5281 ctcaggtgtt caatttcatg ttctagttgc tttgttttac tggtttcacc tgttctatta
     5341 ggtgttacat gctgttcatc tgttacattg tcgatctgtt catggtgaac agctttgaat
     5401 gcaccaaaaa ctcgtaaaag ctctgatgta tctatctttt ttacaccgtt ttcatctgtg
     5461 catatggaca gttttccctt tgatatgtaa cggtgaacag ttgttctact tttgtttgtt
     5521 agtcttgatg cttcactgat agatacaaga gccataagaa cctcagatcc ttccgtattt
     5581 agccagtatg ttctctagtg tggttcgttg tttttgcgtg agccatgaga acgaaccatt
     5641 gagatcatac ttactttgca tgtcactcaa aaattttgcc tcaaaactgg tgagctgaat
     5701 ttttgcagtt aaagcatcgt gtagtgtttt tcttagtccg ttatgtaggt aggaatctga
     5761 tgtaatggtt gttggtattt tgtcaccatt catttttatc tggttgttct caagttcggt
     5821 tacgagatcc atttgtctat ctagttcaac ttggaaaatc
//
