LOCUS       sequence-454156-        5943 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  recA 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 5943)
  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 5943)
  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--PgadX-YFP
FEATURES             Location/Qualifiers
     source          1..5943
                     /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"
     CDS             2240..2953
                     /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"
     terminator      2994..3080
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(3268..3339)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(4728..5678)
                     /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(5726..5943,1..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 tgcaccatca gatccttggc ggcaagaaag ccatccagtt
     1501 tactttgcag ggcttcccaa ccttaccaga gggcgcccca gctggcaatt ccgacgtcta
     1561 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     1621 tcttcacctc gatggatcct gctggaagac tacaaagcct ccctgaaata tctcagcgat
     1681 cacccgaaac tgcagggtat tgcccagcag aacagcttta aacacacctg ataacataac
     1741 gttgtaaaaa ccgaatgccc agcctttaaa aaaacagctg ggcattcggt tgcttattaa
     1801 tcgcaataat atattggctg tctattcatc gtgttgataa gatatatgtt atatgaatgt
     1861 ttattaaatg tagcgattac ctttaatgtc acgaaaaggg attatttgct tactattaat
     1921 ttccctgtgt tcttaaacac gtttttgtat tataaatcca gtcatcctgc ccgtagaaca
     1981 tcacacatta tcatcctgtt ctcccgcttc gtttaaattt atttatcaat caatttgact
     2041 taagagggcg gcgtgctaca ttaataaaca gtaatatgtt tatgtaatat taagtcaact
     2101 atgcaatcac tacatgggaa ttgtctaatt gcgtatgcaa gacataaata tattctcacc
     2161 atggttaatg gtgaatatcg ctattttaat ggcggtgacc tctcgagaga tcctctagat
     2221 ttaagaagga gatatacata tgagtaaagg agaagaactt ttcactggag ttgtcccaat
     2281 tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga
     2341 aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc
     2401 tgttccatgg ccaacacttg tcactacttt cggttatggt cttcaatgct ttgcgagata
     2461 cccagatcat atgaaacagc atgacttttt caagagtgcc atgcccgaag gttatgtaca
     2521 ggaaagaact atatttttca aagatgacgg gaactacaag acacgtgctg aagtcaagtt
     2581 tgaaggtgat acccttgtta atagaatcga gttaaaaggt attgatttta aagaagatgg
     2641 aaacattctt ggacacaaat tggaatacaa ctataactca cacaatgtat acatcatggc
     2701 agacaaacaa aagaatggaa tcaaagttaa cttcaaaatt agacacaaca ttgaagatgg
     2761 aagcgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct
     2821 tttaccagac aaccattacc tgtcctatca atctgccctt tcgaaagatc ccaacgaaaa
     2881 gagagaccac atggtccttc ttgagtttgt aacagctgct gggattacac atggcatgga
     2941 tgaactatac aaataaatgt ccagacctgc aggcatgcaa gctctagagg catcaaataa
     3001 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
     3061 ctctcctgag taggacaaat ccgccgccct agacctaggg tacgggtttt gctgcccgca
     3121 aacgggctgt tctggtgttg ctagtttgtt atcagaatcg cagatccggc ttcagccggt
     3181 ttgccggctg aaagcgctat ttcttccaga attgccatga ttttttcccc acgggaggcg
     3241 tcactggctc ccgtgttgtc ggcagctgag agcgttcacc gacaaacaac agataaaacg
     3301 aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctctaga gcttgcatgc
     3361 ctgcaggtct ggacatttat ttatacagtt catccatgcc cagggtaata ccagcagcgg
     3421 ttacaaactc cagcaggacc atgtgatcgc gcttttcgtt cggatcttta gacagacgag
     3481 attgggtgga caggtaatgg ttgtctggca gcaggacagg gccatcgcca attggagtat
     3541 tttgttgata atggtctgcc agttgaacgc taccatcttc aatgttgtga cgaattttga
     3601 aattagcttt gataccattc ttttgtttgt cagcagtgat gtatacatta tcagaaatag
     3661 cgttgtattc cagtttgtga cccagaatgt taccatcttc tttaaaatca atacctttca
     3721 gctcgatacg attaaccagg gtatcacctt caaacttgac ttcagcacgg gtcttgtagt
     3781 taccgtcatc tttgaaaaag atagtacgtt cctgaacata accttccggc atggcagact
     3841 tgaaaaagtc atgctgtttc atgtgatctg ggtaacgagc aaagcattga acaccccaag
     3901 tcagagtagt gaccagggtt ggccacggaa caggcagttt accagtagtg caaataaatt
     3961 tcagggtcag tttaccgtag gttgcatcac cttcaccctc accagagaca gaaaatttgt
     4021 gaccattaac atcaccatcc agttcaacca gaattggaac aacaccagtg aacagttctt
     4081 cacctttgct tcttttgcta gtcatatgta tatctccttc ttaaatctag aggatctctc
     4141 gagcgcatga tggagccttt accaaattgt ttctcaatct ggcccagtgc tgccgccaac
     4201 gctttctgtt tgttttcgtc gatagccatt tttactcctg tcatgccggg taataccgga
     4261 tagtcaatat gttctgttga agcaattata ctgtatgctc atacagtatc aagtgttttg
     4321 tagaaattgt tgccacaagg tctgcaagga tccatcgagg tgaagacgaa agggcctttg
     4381 attcgataag cagcatcgcc tgtttcaggc tgtctatgtg tgactgttga gctgtaacaa
     4441 gttgtctcag gtgttcaatt tcatgttcta gttgctttgt tttactggtt tcacctgttc
     4501 tattaggtgt tacatgctgt tcatctgtta cattgtcgat ctgttcatgg tgaacagctt
     4561 tgaatgcacc aaaaactcgt aaaagctctg atgtatctat cttttttaca ccgttttcat
     4621 ctgtgcatat ggacagtttt ccctttgata tgtaacggtg aacagttgtt ctacttttgt
     4681 ttgttagtct tgatgcttca ctgatagata caagagccat aagaacctca gatccttccg
     4741 tatttagcca gtatgttctc tagtgtggtt cgttgttttt gcgtgagcca tgagaacgaa
     4801 ccattgagat catacttact ttgcatgtca ctcaaaaatt ttgcctcaaa actggtgagc
     4861 tgaatttttg cagttaaagc atcgtgtagt gtttttctta gtccgttatg taggtaggaa
     4921 tctgatgtaa tggttgttgg tattttgtca ccattcattt ttatctggtt gttctcaagt
     4981 tcggttacga gatccatttg tctatctagt tcaacttgga aaatcaacgt atcagtcggg
     5041 cggcctcgct tatcaaccac caatttcata ttgctgtaag tgtttaaatc tttacttatt
     5101 ggtttcaaaa cccattggtt aagcctttta aactcatggt agttattttc aagcattaac
     5161 atgaacttaa attcatcaag gctaatctct atatttgcct tgtgagtttt cttttgtgtt
     5221 agttctttta ataaccactc ataaatcctc atagagtatt tgttttcaaa agacttaaca
     5281 tgttccagat tatattttat gaattttttt aactggaaaa gataaggcaa tatctcttca
     5341 ctaaaaacta attctaattt ttcgcttgag aacttggcat agtttgtcca ctggaaaatc
     5401 tcaaagcctt taaccaaagg attcctgatt tccacagttc tcgtcatcag ctctctggtt
     5461 gctttagcta atacaccata agcattttcc ctactgatgt tcatcatctg agcgtattgg
     5521 ttataagtga acgataccgt ccgttctttc cttgtagggt tttcaatcgt ggggttgagt
     5581 agtgccacac agcataaaat tagcttggtt tcatgctccg ttaagtcata gcgactaatc
     5641 gctagttcat ttgctttgaa aacaactaat tcagacatac atctcaattg gtctaggtga
     5701 ttttaatcac tataccaatt gagatgggct agtcaatgat aattactagt ccttttcctt
     5761 tgagttgtgg gtatctgtaa attctgctag acctttgctg gaaaacttgt aaattctgct
     5821 agaccctctg taaattccgc tagacctttg tgtgtttttt ttgtttatat tcaagtggtt
     5881 ataatttata gaataaagaa agaataaaaa aagataaaaa gaatagatcc cagccctgtg
     5941 tat
//
