LOCUS       sequence-454155-        5636 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  recA promoter driving expression of CFP reporter and constitutive 
            promoter driving expression of YFP reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5636)
  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 5636)
  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--Pconst-YFP
FEATURES             Location/Qualifiers
     source          1..5636
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(681..775)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(806..1600)
                     /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(917..936)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1527..1546
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1779..1797
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      2832..2918
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(3106..3177)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3218..3931)
                     /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"
     primer_bind     complement(4109..4128)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4605..5555)
                     /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(5603..5636,1..189)
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 ttgagttgtg ggtatctgta aattctgcta gacctttgct ggaaaacttg taaattctgc
       61 tagaccctct gtaaattccg ctagaccttt gtgtgttttt tttgtttata ttcaagtggt
      121 tataatttat agaataaaga aagaataaaa aaagataaaa agaatagatc ccagccctgt
      181 gtataactca ctactttagt cagttccgca gtattacaaa aggatgtcgc aaacgctgtt
      241 tgctcctcta caaaacagac cttaaaaccc taaaggctta agtagcaccc tcgcaagctc
      301 gggcaaatcg ctgaatattc cttttgtctc cgaccatcag gcacctgagt cgctgtcttt
      361 ttcgtgacat tcagttcgct gcgctcacgg ctctggcagt gaatgggggt aaatggcact
      421 acaggcgcct tttatggatt catgcaagga aactacccat aatacaagaa aagcccgtca
      481 cgggcttctc agggcgtttt atggcgggtc tgctatgtgg tgctatctga ctttttgctg
      541 ttcagcagtt cctgccctct gattttccag tctgaccact tcggattatc ccgtgacagg
      601 tcattcagac tggctaatgc acccagtaag gcagcggtat catcaacagg cttacccgtc
      661 ttactgtccc tagtgcttgg attctcacca ataaaaaacg cccggcggca accgagcgtt
      721 ctgaacaaat ccagatggag ttctgaggtc attactggat ctatcaacag gagtccaagc
      781 gagctctcga accccagagt cccgctcaga agaactcgtc aagaaggcga tagaaggcga
      841 tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc
      901 cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca
      961 cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg
     1021 gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga
     1081 gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat
     1141 cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt
     1201 cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg
     1261 atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca
     1321 atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc
     1381 ccgtcgtggc cagccacgat agccgcgctg cctcgtcctg cagttcattc agggcaccgg
     1441 acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg
     1501 catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag
     1561 cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg
     1621 tctcttgatc agatcttgat cccctgcgcc atcagatcct tggcggcaag aaagccatcc
     1681 agtttacttt gcagggcttc ccaaccttac cagagggcgc cccagctggc aattccgacg
     1741 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     1801 ttcgtcttca cctcgatgga tccttgcaga ccttgtggca acaatttcta caaaacactt
     1861 gatactgtat gagcatacag tataattgct tcaacagaac atattgacta tccggtatta
     1921 cccggcatga caggagtaaa aatggctatc gacgaaaaca aacagaaagc gttggcggca
     1981 gcactgggcc agattgagaa acaatttggt aaaggctcca tcatgcgctc gagagatcct
     2041 ctagatttaa gaaggagata tacatatgac tagcaaaaga agcaaaggtg aagaactgtt
     2101 cactggtgtt gttccaattc tggttgaact ggatggtgat gttaatggtc acaaattttc
     2161 tgtctctggt gagggtgaag gtgatgcaac ctacggtaaa ctgaccctga aatttatttg
     2221 cactactggt aaactgcctg ttccgtggcc aaccctggtc actactctga cttggggtgt
     2281 tcaatgcttt gctcgttacc cagatcacat gaaacagcat gactttttca agtctgccat
     2341 gccggaaggt tatgttcagg aacgtactat ctttttcaaa gatgacggta actacaagac
     2401 ccgtgctgaa gtcaagtttg aaggtgatac cctggttaat cgtatcgagc tgaaaggtat
     2461 tgattttaaa gaagatggta acattctggg tcacaaactg gaatacaacg ctatttctga
     2521 taatgtatac atcactgctg acaaacaaaa gaatggtatc aaagctaatt tcaaaattcg
     2581 tcacaacatt gaagatggta gcgttcaact ggcagaccat tatcaacaaa atactccaat
     2641 tggcgatggc cctgtcctgc tgccagacaa ccattacctg tccacccaat ctcgtctgtc
     2701 taaagatccg aacgaaaagc gcgatcacat ggtcctgctg gagtttgtaa ccgctgctgg
     2761 tattaccctg ggcatggatg aactgtataa ataaatgtcc agacctgcag gcatgcaagc
     2821 tctagaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     2881 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgccctag acctagggta
     2941 cgggttttgc tgcccgcaaa cgggctgttc tggtgttgct agtttgttat cagaatcgca
     3001 gatccggctt cagccggttt gccggctgaa agcgctattt cttccagaat tgccatgatt
     3061 ttttccccac gggaggcgtc actggctccc gtgttgtcgg cagctgagag cgttcaccga
     3121 caaacaacag ataaaacgaa aggcccagtc tttcgactga gcctttcgtt ttatttgatg
     3181 cctctagagc ttgcatgcct gcaggtctgg acatttattt gtatagttca tccatgccat
     3241 gtgtaatccc agcagctgtt acaaactcaa gaaggaccat gtggtctctc ttttcgttgg
     3301 gatctttcga aagggcagat tgataggaca ggtaatggtt gtctggtaaa aggacagggc
     3361 catcgccaat tggagtattt tgttgataat ggtctgctag ttgaacgctt ccatcttcaa
     3421 tgttgtgtct aattttgaag ttaactttga ttccattctt ttgtttgtct gccatgatgt
     3481 atacattgtg tgagttatag ttgtattcca atttgtgtcc aagaatgttt ccatcttctt
     3541 taaaatcaat accttttaac tcgattctat taacaagggt atcaccttca aacttgactt
     3601 cagcacgtgt cttgtagttc ccgtcatctt tgaaaaatat agttctttcc tgtacataac
     3661 cttcgggcat ggcactcttg aaaaagtcat gctgtttcat atgatctggg tatctcgcaa
     3721 agcattgaag accataaccg aaagtagtga caagtgttgg ccatggaaca ggtagttttc
     3781 cagtagtgca aataaattta agggtaagtt ttccgtatgt tgcatcacct tcaccctctc
     3841 cactgacaga aaatttgtgc ccattaacat caccatctaa ttcaacaaga attgggacaa
     3901 ctccagtgaa aagttcttct cctttactca tatgtatatc tccttcttaa atctagagga
     3961 tcccctcgag gtttcgccgt gtccctctcg atggctgtaa ttttcctata ggttagactt
     4021 taagacaata ctctttttga taactcgatc ctctacgccg gacgcatcgt ggccggcatc
     4081 accggcgcca caggtgcggt tgctggcgcc tatatcgccg acatcaccga tggggaagat
     4141 cgggctcgcc acttcgggct catgagcgct tgtttcggcg tgggtatggt ggcaggcccc
     4201 gtggccgggg gactgttggg cgccatctcc ttgcatgcga agacgaaagg gcctttgatt
     4261 cgataagcag catcgcctgt ttcaggctgt ctatgtgtga ctgttgagct gtaacaagtt
     4321 gtctcaggtg ttcaatttca tgttctagtt gctttgtttt actggtttca cctgttctat
     4381 taggtgttac atgctgttca tctgttacat tgtcgatctg ttcatggtga acagctttga
     4441 atgcaccaaa aactcgtaaa agctctgatg tatctatctt ttttacaccg ttttcatctg
     4501 tgcatatgga cagttttccc tttgatatgt aacggtgaac agttgttcta cttttgtttg
     4561 ttagtcttga tgcttcactg atagatacaa gagccataag aacctcagat ccttccgtat
     4621 ttagccagta tgttctctag tgtggttcgt tgtttttgcg tgagccatga gaacgaacca
     4681 ttgagatcat acttactttg catgtcactc aaaaattttg cctcaaaact ggtgagctga
     4741 atttttgcag ttaaagcatc gtgtagtgtt tttcttagtc cgttatgtag gtaggaatct
     4801 gatgtaatgg ttgttggtat tttgtcacca ttcattttta tctggttgtt ctcaagttcg
     4861 gttacgagat ccatttgtct atctagttca acttggaaaa tcaacgtatc agtcgggcgg
     4921 cctcgcttat caaccaccaa tttcatattg ctgtaagtgt ttaaatcttt acttattggt
     4981 ttcaaaaccc attggttaag ccttttaaac tcatggtagt tattttcaag cattaacatg
     5041 aacttaaatt catcaaggct aatctctata tttgccttgt gagttttctt ttgtgttagt
     5101 tcttttaata accactcata aatcctcata gagtatttgt tttcaaaaga cttaacatgt
     5161 tccagattat attttatgaa tttttttaac tggaaaagat aaggcaatat ctcttcacta
     5221 aaaactaatt ctaatttttc gcttgagaac ttggcatagt ttgtccactg gaaaatctca
     5281 aagcctttaa ccaaaggatt cctgatttcc acagttctcg tcatcagctc tctggttgct
     5341 ttagctaata caccataagc attttcccta ctgatgttca tcatctgagc gtattggtta
     5401 taagtgaacg ataccgtccg ttctttcctt gtagggtttt caatcgtggg gttgagtagt
     5461 gccacacagc ataaaattag cttggtttca tgctccgtta agtcatagcg actaatcgct
     5521 agttcatttg ctttgaaaac aactaattca gacatacatc tcaattggtc taggtgattt
     5581 taatcactat accaattgag atgggctagt caatgataat tactagtcct tttcct
//
