LOCUS       sequence-454184-        5687 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive 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 5687)
  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 5687)
  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--Pconst-YFP
FEATURES             Location/Qualifiers
     source          1..5687
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      48..270
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(762..856)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(887..1681)
                     /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(998..1017)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1608..1627
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1860..1878
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      2964..3050
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(3238..3309)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3350..4063)
                     /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(4737..5687)
                     /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 acatctcaat tggtctaggt gattttaatc actataccaa ttgagatggg ctagtcaatg
       61 ataattacta gtccttttcc tttgagttgt gggtatctgt aaattctgct agacctttgc
      121 tggaaaactt gtaaattctg ctagaccctc tgtaaattcc gctagacctt tgtgtgtttt
      181 ttttgtttat attcaagtgg ttataattta tagaataaag aaagaataaa aaaagataaa
      241 aagaatagat cccagccctg tgtataactc actactttag tcagttccgc agtattacaa
      301 aaggatgtcg caaacgctgt ttgctcctct acaaaacaga ccttaaaacc ctaaaggctt
      361 aagtagcacc ctcgcaagct cgggcaaatc gctgaatatt ccttttgtct ccgaccatca
      421 ggcacctgag tcgctgtctt tttcgtgaca ttcagttcgc tgcgctcacg gctctggcag
      481 tgaatggggg taaatggcac tacaggcgcc ttttatggat tcatgcaagg aaactaccca
      541 taatacaaga aaagcccgtc acgggcttct cagggcgttt tatggcgggt ctgctatgtg
      601 gtgctatctg actttttgct gttcagcagt tcctgccctc tgattttcca gtctgaccac
      661 ttcggattat cccgtgacag gtcattcaga ctggctaatg cacccagtaa ggcagcggta
      721 tcatcaacag gcttacccgt cttactgtcc ctagtgcttg gattctcacc aataaaaaac
      781 gcccggcggc aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga
      841 tctatcaaca ggagtccaag cgagctctcg aaccccagag tcccgctcag aagaactcgt
      901 caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga
      961 ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta
     1021 tgtcctgata gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc
     1081 cattttccac catgatattc ggcaagcagg catcgccatg ggtcacgacg agatcctcgc
     1141 cgtcgggcat gcgcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct
     1201 cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga
     1261 tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc
     1321 gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat
     1381 cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga
     1441 gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct
     1501 gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg
     1561 ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc
     1621 cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca
     1681 tgcgaaacga tcctcatcct gtctcttgat cagatcttga tcccctgcgc catcagatcc
     1741 ttggcggcaa gaaagccatc cagtttactt tgcagggctt cccaacctta ccagagggcg
     1801 ccccagctgg caattccgac gtctaagaaa ccattattat catgacatta acctataaaa
     1861 ataggcgtat cacgaggccc tttcgtcttc gcatgcaagg agatggcgcc caacagtccc
     1921 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
     1981 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
     2041 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagttatc aaaaagagta
     2101 ttgtcttaaa gtctaaccta taggaaaatt acagccatcg agagggacac ggcgaaacct
     2161 cgaggggatc ctctagattt aagaaggaga tatacatatg actagcaaaa gaagcaaagg
     2221 tgaagaactg ttcactggtg ttgttccaat tctggttgaa ctggatggtg atgttaatgg
     2281 tcacaaattt tctgtctctg gtgagggtga aggtgatgca acctacggta aactgaccct
     2341 gaaatttatt tgcactactg gtaaactgcc tgttccgtgg ccaaccctgg tcactactct
     2401 gacttggggt gttcaatgct ttgctcgtta cccagatcac atgaaacagc atgacttttt
     2461 caagtctgcc atgccggaag gttatgttca ggaacgtact atctttttca aagatgacgg
     2521 taactacaag acccgtgctg aagtcaagtt tgaaggtgat accctggtta atcgtatcga
     2581 gctgaaaggt attgatttta aagaagatgg taacattctg ggtcacaaac tggaatacaa
     2641 cgctatttct gataatgtat acatcactgc tgacaaacaa aagaatggta tcaaagctaa
     2701 tttcaaaatt cgtcacaaca ttgaagatgg tagcgttcaa ctggcagacc attatcaaca
     2761 aaatactcca attggcgatg gccctgtcct gctgccagac aaccattacc tgtccaccca
     2821 atctcgtctg tctaaagatc cgaacgaaaa gcgcgatcac atggtcctgc tggagtttgt
     2881 aaccgctgct ggtattaccc tgggcatgga tgaactgtat aaataaatgt ccagacctgc
     2941 aggcatgcaa gctctagagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     3001 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgccct
     3061 agacctaggg tacgggtttt gctgcccgca aacgggctgt tctggtgttg ctagtttgtt
     3121 atcagaatcg cagatccggc ttcagccggt ttgccggctg aaagcgctat ttcttccaga
     3181 attgccatga ttttttcccc acgggaggcg tcactggctc ccgtgttgtc ggcagctgag
     3241 agcgttcacc gacaaacaac agataaaacg aaaggcccag tctttcgact gagcctttcg
     3301 ttttatttga tgcctctaga gcttgcatgc ctgcaggtct ggacatttat ttgtatagtt
     3361 catccatgcc atgtgtaatc ccagcagctg ttacaaactc aagaaggacc atgtggtctc
     3421 tcttttcgtt gggatctttc gaaagggcag attgatagga caggtaatgg ttgtctggta
     3481 aaaggacagg gccatcgcca attggagtat tttgttgata atggtctgct agttgaacgc
     3541 ttccatcttc aatgttgtgt ctaattttga agttaacttt gattccattc ttttgtttgt
     3601 ctgccatgat gtatacattg tgtgagttat agttgtattc caatttgtgt ccaagaatgt
     3661 ttccatcttc tttaaaatca atacctttta actcgattct attaacaagg gtatcacctt
     3721 caaacttgac ttcagcacgt gtcttgtagt tcccgtcatc tttgaaaaat atagttcttt
     3781 cctgtacata accttcgggc atggcactct tgaaaaagtc atgctgtttc atatgatctg
     3841 ggtatctcgc aaagcattga agaccataac cgaaagtagt gacaagtgtt ggccatggaa
     3901 caggtagttt tccagtagtg caaataaatt taagggtaag ttttccgtat gttgcatcac
     3961 cttcaccctc tccactgaca gaaaatttgt gcccattaac atcaccatct aattcaacaa
     4021 gaattgggac aactccagtg aaaagttctt ctcctttact catatgtata tctccttctt
     4081 aaatctagag gatcccctcg aggtttcgcc gtgtccctct cgatggctgt aattttccta
     4141 taggttagac tttaagacaa tactcttttt gataactcga tcctctacgc cggacgcatc
     4201 gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc
     4261 gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg
     4321 gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc gaagacgaaa
     4381 gggcctttga ttcgataagc agcatcgcct gtttcaggct gtctatgtgt gactgttgag
     4441 ctgtaacaag ttgtctcagg tgttcaattt catgttctag ttgctttgtt ttactggttt
     4501 cacctgttct attaggtgtt acatgctgtt catctgttac attgtcgatc tgttcatggt
     4561 gaacagcttt gaatgcacca aaaactcgta aaagctctga tgtatctatc ttttttacac
     4621 cgttttcatc tgtgcatatg gacagttttc cctttgatat gtaacggtga acagttgttc
     4681 tacttttgtt tgttagtctt gatgcttcac tgatagatac aagagccata agaacctcag
     4741 atccttccgt atttagccag tatgttctct agtgtggttc gttgtttttg cgtgagccat
     4801 gagaacgaac cattgagatc atacttactt tgcatgtcac tcaaaaattt tgcctcaaaa
     4861 ctggtgagct gaatttttgc agttaaagca tcgtgtagtg tttttcttag tccgttatgt
     4921 aggtaggaat ctgatgtaat ggttgttggt attttgtcac cattcatttt tatctggttg
     4981 ttctcaagtt cggttacgag atccatttgt ctatctagtt caacttggaa aatcaacgta
     5041 tcagtcgggc ggcctcgctt atcaaccacc aatttcatat tgctgtaagt gtttaaatct
     5101 ttacttattg gtttcaaaac ccattggtta agccttttaa actcatggta gttattttca
     5161 agcattaaca tgaacttaaa ttcatcaagg ctaatctcta tatttgcctt gtgagttttc
     5221 ttttgtgtta gttcttttaa taaccactca taaatcctca tagagtattt gttttcaaaa
     5281 gacttaacat gttccagatt atattttatg aattttttta actggaaaag ataaggcaat
     5341 atctcttcac taaaaactaa ttctaatttt tcgcttgaga acttggcata gtttgtccac
     5401 tggaaaatct caaagccttt aaccaaagga ttcctgattt ccacagttct cgtcatcagc
     5461 tctctggttg ctttagctaa tacaccataa gcattttccc tactgatgtt catcatctga
     5521 gcgtattggt tataagtgaa cgataccgtc cgttctttcc ttgtagggtt ttcaatcgtg
     5581 gggttgagta gtgccacaca gcataaaatt agcttggttt catgctccgt taagtcatag
     5641 cgactaatcg ctagttcatt tgctttgaaa acaactaatt cagacat
//
