LOCUS       sequence-454099-        5906 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gadX 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 5906)
  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 5906)
  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: PgadX-CFP--Pconst-YFP
FEATURES             Location/Qualifiers
     source          1..5906
                     /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      3190..3276
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(3464..3535)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3576..4289)
                     /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(4467..4486)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4875..5825)
                     /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(5873..5906,1..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 tcctgctgga agactacaaa gcctccctga aatatctcag
     1861 cgatcacccg aaactgcagg gtattgccca gcagaacagc tttaaacaca cctgataaca
     1921 taacgttgta aaaaccgaat gcccagcctt taaaaaaaca gctgggcatt cggttgctta
     1981 ttaatcgcaa taatatattg gctgtctatt catcgtgttg ataagatata tgttatatga
     2041 atgtttatta aatgtagcga ttacctttaa tgtcacgaaa agggattatt tgcttactat
     2101 taatttccct gtgttcttaa acacgttttt gtattataaa tccagtcatc ctgcccgtag
     2161 aacatcacac attatcatcc tgttctcccg cttcgtttaa atttatttat caatcaattt
     2221 gacttaagag ggcggcgtgc tacattaata aacagtaata tgtttatgta atattaagtc
     2281 aactatgcaa tcactacatg ggaattgtct aattgcgtat gcaagacata aatatattct
     2341 caccatggtt aatggtgaat atcgctattt taatggcggt gacctctcga gagatcctct
     2401 agatttaaga aggagatata catatgacta gcaaaagaag caaaggtgaa gaactgttca
     2461 ctggtgttgt tccaattctg gttgaactgg atggtgatgt taatggtcac aaattttctg
     2521 tctctggtga gggtgaaggt gatgcaacct acggtaaact gaccctgaaa tttatttgca
     2581 ctactggtaa actgcctgtt ccgtggccaa ccctggtcac tactctgact tggggtgttc
     2641 aatgctttgc tcgttaccca gatcacatga aacagcatga ctttttcaag tctgccatgc
     2701 cggaaggtta tgttcaggaa cgtactatct ttttcaaaga tgacggtaac tacaagaccc
     2761 gtgctgaagt caagtttgaa ggtgataccc tggttaatcg tatcgagctg aaaggtattg
     2821 attttaaaga agatggtaac attctgggtc acaaactgga atacaacgct atttctgata
     2881 atgtatacat cactgctgac aaacaaaaga atggtatcaa agctaatttc aaaattcgtc
     2941 acaacattga agatggtagc gttcaactgg cagaccatta tcaacaaaat actccaattg
     3001 gcgatggccc tgtcctgctg ccagacaacc attacctgtc cacccaatct cgtctgtcta
     3061 aagatccgaa cgaaaagcgc gatcacatgg tcctgctgga gtttgtaacc gctgctggta
     3121 ttaccctggg catggatgaa ctgtataaat aaatgtccag acctgcaggc atgcaagctc
     3181 tagaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     3241 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgccctagac ctagggtacg
     3301 ggttttgctg cccgcaaacg ggctgttctg gtgttgctag tttgttatca gaatcgcaga
     3361 tccggcttca gccggtttgc cggctgaaag cgctatttct tccagaattg ccatgatttt
     3421 ttccccacgg gaggcgtcac tggctcccgt gttgtcggca gctgagagcg ttcaccgaca
     3481 aacaacagat aaaacgaaag gcccagtctt tcgactgagc ctttcgtttt atttgatgcc
     3541 tctagagctt gcatgcctgc aggtctggac atttatttgt atagttcatc catgccatgt
     3601 gtaatcccag cagctgttac aaactcaaga aggaccatgt ggtctctctt ttcgttggga
     3661 tctttcgaaa gggcagattg ataggacagg taatggttgt ctggtaaaag gacagggcca
     3721 tcgccaattg gagtattttg ttgataatgg tctgctagtt gaacgcttcc atcttcaatg
     3781 ttgtgtctaa ttttgaagtt aactttgatt ccattctttt gtttgtctgc catgatgtat
     3841 acattgtgtg agttatagtt gtattccaat ttgtgtccaa gaatgtttcc atcttcttta
     3901 aaatcaatac cttttaactc gattctatta acaagggtat caccttcaaa cttgacttca
     3961 gcacgtgtct tgtagttccc gtcatctttg aaaaatatag ttctttcctg tacataacct
     4021 tcgggcatgg cactcttgaa aaagtcatgc tgtttcatat gatctgggta tctcgcaaag
     4081 cattgaagac cataaccgaa agtagtgaca agtgttggcc atggaacagg tagttttcca
     4141 gtagtgcaaa taaatttaag ggtaagtttt ccgtatgttg catcaccttc accctctcca
     4201 ctgacagaaa atttgtgccc attaacatca ccatctaatt caacaagaat tgggacaact
     4261 ccagtgaaaa gttcttctcc tttactcata tgtatatctc cttcttaaat ctagaggatc
     4321 ccctcgaggt ttcgccgtgt ccctctcgat ggctgtaatt ttcctatagg ttagacttta
     4381 agacaatact ctttttgata actcgatcct ctacgccgga cgcatcgtgg ccggcatcac
     4441 cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg
     4501 ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt
     4561 ggccggggga ctgttgggct gtaacaagtt gtctcaggtg ttcaatttca tgttctagtt
     4621 gctttgtttt actggtttca cctgttctat taggtgttac atgctgttca tctgttacat
     4681 tgtcgatctg ttcatggtga acagctttga atgcaccaaa aactcgtaaa agctctgatg
     4741 tatctatctt ttttacaccg ttttcatctg tgcatatgga cagttttccc tttgatatgt
     4801 aacggtgaac agttgttcta cttttgtttg ttagtcttga tgcttcactg atagatacaa
     4861 gagccataag aacctcagat ccttccgtat ttagccagta tgttctctag tgtggttcgt
     4921 tgtttttgcg tgagccatga gaacgaacca ttgagatcat acttactttg catgtcactc
     4981 aaaaattttg cctcaaaact ggtgagctga atttttgcag ttaaagcatc gtgtagtgtt
     5041 tttcttagtc cgttatgtag gtaggaatct gatgtaatgg ttgttggtat tttgtcacca
     5101 ttcattttta tctggttgtt ctcaagttcg gttacgagat ccatttgtct atctagttca
     5161 acttggaaaa tcaacgtatc agtcgggcgg cctcgcttat caaccaccaa tttcatattg
     5221 ctgtaagtgt ttaaatcttt acttattggt ttcaaaaccc attggttaag ccttttaaac
     5281 tcatggtagt tattttcaag cattaacatg aacttaaatt catcaaggct aatctctata
     5341 tttgccttgt gagttttctt ttgtgttagt tcttttaata accactcata aatcctcata
     5401 gagtatttgt tttcaaaaga cttaacatgt tccagattat attttatgaa tttttttaac
     5461 tggaaaagat aaggcaatat ctcttcacta aaaactaatt ctaatttttc gcttgagaac
     5521 ttggcatagt ttgtccactg gaaaatctca aagcctttaa ccaaaggatt cctgatttcc
     5581 acagttctcg tcatcagctc tctggttgct ttagctaata caccataagc attttcccta
     5641 ctgatgttca tcatctgagc gtattggtta taagtgaacg ataccgtccg ttctttcctt
     5701 gtagggtttt caatcgtggg gttgagtagt gccacacagc ataaaattag cttggtttca
     5761 tgctccgtta agtcatagcg actaatcgct agttcatttg ctttgaaaac aactaattca
     5821 gacatacatc tcaattggtc taggtgattt taatcactat accaattgag atgggctagt
     5881 caatgataat tactagtcct tttcct
//
