LOCUS       sequence-454109-        5570 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  recA 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 5570)
  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 5570)
  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: PrecA-CFP--PrecA-YFP
FEATURES             Location/Qualifiers
     source          1..5570
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(497..591)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(622..1416)
                     /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(733..752)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1343..1362
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1595..1613
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      2648..2734
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(2922..2993)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3034..3747)
                     /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(4355..5305)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      join(5353..5570,1..5)
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 aactcactac tttagtcagt tccgcagtat tacaaaagga tgtcgcaaac gctgtttgct
       61 cctctacaaa acagacctta aaaccctaaa ggcttaagta gcaccctcgc aagctcgggc
      121 aaatcgctga atattccttt tgtctccgac catcaggcac ctgagtcgct gtctttttcg
      181 tgacattcag ttcgctgcgc tcacggctct ggcagtgaat gggggtaaat ggcactacag
      241 gcgcctttta tggattcatg caaggaaact acccataata caagaaaagc ccgtcacggg
      301 cttctcaggg cgttttatgg cgggtctgct atgtggtgct atctgacttt ttgctgttca
      361 gcagttcctg ccctctgatt ttccagtctg accacttcgg attatcccgt gacaggtcat
      421 tcagactggc taatgcaccc agtaaggcag cggtatcatc aacaggctta cccgtcttac
      481 tgtccctagt gcttggattc tcaccaataa aaaacgcccg gcggcaaccg agcgttctga
      541 acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt ccaagcgagc
      601 tctcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg
      661 ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc
      721 aagctcttca gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc
      781 cagccggcca cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa
      841 gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg ggcatgcgcg ccttgagcct
      901 ggcgaacagt tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac
      961 aagaccggct tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa
     1021 tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac
     1081 tttctcggca ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag
     1141 cagccagtcc cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt
     1201 cgtggccagc cacgatagcc gcgctgcctc gtcctgcagt tcattcaggg caccggacag
     1261 gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc
     1321 agagcagccg attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc
     1381 cggagaacct gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc
     1441 ttgatcagat cttgatcccc tgcgccatca gatccttggc ggcaagaaag ccatccagtt
     1501 tactttgcag ggcttcccaa ccttaccaga gggcgcccca gctggcaatt ccgacgtcta
     1561 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     1621 tcttcacctc gatggatcct tgcagacctt gtggcaacaa tttctacaaa acacttgata
     1681 ctgtatgagc atacagtata attgcttcaa cagaacatat tgactatccg gtattacccg
     1741 gcatgacagg agtaaaaatg gctatcgacg aaaacaaaca gaaagcgttg gcggcagcac
     1801 tgggccagat tgagaaacaa tttggtaaag gctccatcat gcgctcgaga gatcctctag
     1861 atttaagaag gagatataca tatgactagc aaaagaagca aaggtgaaga actgttcact
     1921 ggtgttgttc caattctggt tgaactggat ggtgatgtta atggtcacaa attttctgtc
     1981 tctggtgagg gtgaaggtga tgcaacctac ggtaaactga ccctgaaatt tatttgcact
     2041 actggtaaac tgcctgttcc gtggccaacc ctggtcacta ctctgacttg gggtgttcaa
     2101 tgctttgctc gttacccaga tcacatgaaa cagcatgact ttttcaagtc tgccatgccg
     2161 gaaggttatg ttcaggaacg tactatcttt ttcaaagatg acggtaacta caagacccgt
     2221 gctgaagtca agtttgaagg tgataccctg gttaatcgta tcgagctgaa aggtattgat
     2281 tttaaagaag atggtaacat tctgggtcac aaactggaat acaacgctat ttctgataat
     2341 gtatacatca ctgctgacaa acaaaagaat ggtatcaaag ctaatttcaa aattcgtcac
     2401 aacattgaag atggtagcgt tcaactggca gaccattatc aacaaaatac tccaattggc
     2461 gatggccctg tcctgctgcc agacaaccat tacctgtcca cccaatctcg tctgtctaaa
     2521 gatccgaacg aaaagcgcga tcacatggtc ctgctggagt ttgtaaccgc tgctggtatt
     2581 accctgggca tggatgaact gtataaataa atgtccagac ctgcaggcat gcaagctcta
     2641 gaggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     2701 tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ccctagacct agggtacggg
     2761 ttttgctgcc cgcaaacggg ctgttctggt gttgctagtt tgttatcaga atcgcagatc
     2821 cggcttcagc cggtttgccg gctgaaagcg ctatttcttc cagaattgcc atgatttttt
     2881 ccccacggga ggcgtcactg gctcccgtgt tgtcggcagc tgagagcgtt caccgacaaa
     2941 caacagataa aacgaaaggc ccagtctttc gactgagcct ttcgttttat ttgatgcctc
     3001 tagagcttgc atgcctgcag gtctggacat ttatttgtat agttcatcca tgccatgtgt
     3061 aatcccagca gctgttacaa actcaagaag gaccatgtgg tctctctttt cgttgggatc
     3121 tttcgaaagg gcagattgat aggacaggta atggttgtct ggtaaaagga cagggccatc
     3181 gccaattgga gtattttgtt gataatggtc tgctagttga acgcttccat cttcaatgtt
     3241 gtgtctaatt ttgaagttaa ctttgattcc attcttttgt ttgtctgcca tgatgtatac
     3301 attgtgtgag ttatagttgt attccaattt gtgtccaaga atgtttccat cttctttaaa
     3361 atcaatacct tttaactcga ttctattaac aagggtatca ccttcaaact tgacttcagc
     3421 acgtgtcttg tagttcccgt catctttgaa aaatatagtt ctttcctgta cataaccttc
     3481 gggcatggca ctcttgaaaa agtcatgctg tttcatatga tctgggtatc tcgcaaagca
     3541 ttgaagacca taaccgaaag tagtgacaag tgttggccat ggaacaggta gttttccagt
     3601 agtgcaaata aatttaaggg taagttttcc gtatgttgca tcaccttcac cctctccact
     3661 gacagaaaat ttgtgcccat taacatcacc atctaattca acaagaattg ggacaactcc
     3721 agtgaaaagt tcttctcctt tactcatatg tatatctcct tcttaaatct agaggatctc
     3781 tcgagcgcat gatggagcct ttaccaaatt gtttctcaat ctggcccagt gctgccgcca
     3841 acgctttctg tttgttttcg tcgatagcca tttttactcc tgtcatgccg ggtaataccg
     3901 gatagtcaat atgttctgtt gaagcaatta tactgtatgc tcatacagta tcaagtgttt
     3961 tgtagaaatt gttgccacaa ggtctgcaag gatccatcga ggtgttgatt cgataagcag
     4021 catcgcctgt ttcaggctgt ctatgtgtga ctgttgagct gtaacaagtt gtctcaggtg
     4081 ttcaatttca tgttctagtt gctttgtttt actggtttca cctgttctat taggtgttac
     4141 atgctgttca tctgttacat tgtcgatctg ttcatggtga acagctttga atgcaccaaa
     4201 aactcgtaaa agctctgatg tatctatctt ttttacaccg ttttcatctg tgcatatgga
     4261 cagttttccc tttgatatgt aacggtgaac agttgttcta cttttgtttg ttagtcttga
     4321 tgcttcactg atagatacaa gagccataag aacctcagat ccttccgtat ttagccagta
     4381 tgttctctag tgtggttcgt tgtttttgcg tgagccatga gaacgaacca ttgagatcat
     4441 acttactttg catgtcactc aaaaattttg cctcaaaact ggtgagctga atttttgcag
     4501 ttaaagcatc gtgtagtgtt tttcttagtc cgttatgtag gtaggaatct gatgtaatgg
     4561 ttgttggtat tttgtcacca ttcattttta tctggttgtt ctcaagttcg gttacgagat
     4621 ccatttgtct atctagttca acttggaaaa tcaacgtatc agtcgggcgg cctcgcttat
     4681 caaccaccaa tttcatattg ctgtaagtgt ttaaatcttt acttattggt ttcaaaaccc
     4741 attggttaag ccttttaaac tcatggtagt tattttcaag cattaacatg aacttaaatt
     4801 catcaaggct aatctctata tttgccttgt gagttttctt ttgtgttagt tcttttaata
     4861 accactcata aatcctcata gagtatttgt tttcaaaaga cttaacatgt tccagattat
     4921 attttatgaa tttttttaac tggaaaagat aaggcaatat ctcttcacta aaaactaatt
     4981 ctaatttttc gcttgagaac ttggcatagt ttgtccactg gaaaatctca aagcctttaa
     5041 ccaaaggatt cctgatttcc acagttctcg tcatcagctc tctggttgct ttagctaata
     5101 caccataagc attttcccta ctgatgttca tcatctgagc gtattggtta taagtgaacg
     5161 ataccgtccg ttctttcctt gtagggtttt caatcgtggg gttgagtagt gccacacagc
     5221 ataaaattag cttggtttca tgctccgtta agtcatagcg actaatcgct agttcatttg
     5281 ctttgaaaac aactaattca gacatacatc tcaattggtc taggtgattt taatcactat
     5341 accaattgag atgggctagt caatgataat tactagtcct tttcctttga gttgtgggta
     5401 tctgtaaatt ctgctagacc tttgctggaa aacttgtaaa ttctgctaga ccctctgtaa
     5461 attccgctag acctttgtgt gttttttttg tttatattca agtggttata atttatagaa
     5521 taaagaaaga ataaaaaaag ataaaaagaa tagatcccag ccctgtgtat
//
