LOCUS       sequence-426759-        5520 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A three-colour stress biosensor for real time analysis of 
            physiological stress, genotoxicity, and cytotoxicity of Escherichia 
            coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5520)
  AUTHORS   Zoheir AE, Sobol MS, Meisch L, Ordonez-Rueda D, Kaster AK, Niemeyer 
            CM, Rabe KS
  TITLE     A three-colour stress biosensor reveals multimodal response in 
            single cells and spatiotemporal dynamics of biofilms.
  JOURNAL   NPJ Biofilms Microbiomes. 2023 Aug 21;9(1):57. doi: 
            10.1038/s41522-023-00424-1.
  PUBMED    37604827
REFERENCE   2  (bases 1 to 5520)
  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: "NPJ 
            Biofilms Microbiomes."; date: "2023-08-21"; volume: "9(1):57. doi"; 
            pages: " 10.1038/s41522-023-00424-1"
COMMENT     Sequence Label: RGB-S reporter
FEATURES             Location/Qualifiers
     source          1..5520
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1140..1151
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1309..1328
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1309..1327
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1332..1343
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      2190..2261
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2277..2304
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             2764..3441
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      3461..3532
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3548..3575
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(3726..3743)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3836..4424)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3916..3935)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4598..5392)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(5393..5497)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtggccgca gataaaaaaa atccttagct ttcgctaagg atgatttctg
      181 taacaactta tcagttcagt ttatgaccca gtttgctcgg cagatcacaa taacgtgcaa
      241 ctgcaacttc atgctgttca acataggttt cgttgtttgc ttctttgatg cgttccagac
      301 gataatccac ataataaacg ccaggcattt tcagattctt tgccggtttt ttgctacgat
      361 aggtggtttt ggcatttgca atcagatggc taccaccaac cagtttcagt gccatatcat
      421 tacgaccttc caggccacca tctgccggat acagtgtttc ggtaaatgct tcccaaccta
      481 aggtcttttt ctgcataacc ggaccattgc tggtgaagtt cacaccgcga attttcacat
      541 tataaatcag acaaccatcc tgcagactgg tatcctgtgt tgcggtcaga acaccaccat
      601 cttcataggt ggtaacacgt tcccaggtaa aaccttccgg aaagctctgt ttgaaaaaat
      661 ccgggatacc ctgggtatga ttgataaagg ttttgctacc atacagaaag ctggttgcca
      721 gaatatcaaa tgcaaacggc agcggaccac cttcaacaac tttaatacgc atggtctggg
      781 tgccttcata cggtttacct tcaccttcgc tggtacattt aaagtggtgg ttatcaacgg
      841 tgccttccat atacagtttc atgtgcatgt tttctttgat cagttcttcg cctttgctaa
      901 ccatgaattt ctccgcgttt ttttcgcatt catctcgcta acttcgctta ttatggggat
      961 cagtttcagg gtttcaaggg aagcactcac attgtcatca atcttcgcaa caaggacctc
     1021 ggcgaaacgc ccttcccaac agttgcgcag cctgaatggc gaatgggagc gcctcggtac
     1081 caaattccag aaaagaggcc tcccgaaagg ggggcctttt ttcgttttgg tccccaatta
     1141 ttgaaggccg ctaacgcggc ctttttttgt ttctggtctc ccccagtgag cgcgacgtaa
     1201 tacgactccc ccagtgagcg cgacgtaata cgactccccc agtgagcgcg acgtaatacg
     1261 actcccccag tgagcgcgac gtaatacgac tccccagtga gcgcgacgta atacgactca
     1321 ctatagggcg aattgaagga aggccgtcaa ggccgcatga atggcagaaa ttcgaagaat
     1381 agttacggct agggttgatc tttgttgtca ctggatgtac tgtacatcca tacagtaact
     1441 cacaggggct ggattgatta tgcgtaaagg tgaagaactg tttaccggtg ttgttccgat
     1501 tctggttgaa ctggatggtg atgttaatgg ccacaaattt tcagttagcg gtgaaggcga
     1561 aggtgatgca acctatggta aactgaccct gaaatttatc tgtaccaccg gcaaactgcc
     1621 ggttccgtgg ccgacactgg ttaccacctt tggttatggt gttcagtgtt ttgcacgtta
     1681 tccggatcat atgaaacagc acgatttttt caaaagcgca atgccggaag gttatgttca
     1741 agaacgtacc atcttcttca aagatgacgg caactataaa acccgtgccg aagttaaatt
     1801 tgaaggtgat accctggtga atcgcattga actgaaaggc atcgatttta aagaggatgg
     1861 taatatcctg ggccacaaac tggaatataa ttataatagc cacaacgtgt acatcatggc
     1921 cgacaaacag aaaaatggca tcaaagtgaa cttcaagatc cgccataata ttgaagatgg
     1981 ttcagttcag ctggccgatc attatcagca gaataccccg attggtgatg gtccggttct
     2041 gctgccggat aatcattatc tgagcaccca gagcgcactg agcaaagatc cgaatgaaaa
     2101 acgtgatcac atggtgctgc tggaatttgt taccgcagca ggtattaccc atggtatgga
     2161 tgaactgtat aaatgataaa gccaggcatc aaataaaacg aaaggctcag tcgaaagact
     2221 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct
     2281 caccttcggg tgggcctttc tgcgtttata tactagactg gcacaggaac gttatccgga
     2341 cgttcagttc caccagaccc gcgagcatta attcttgcct ccagggcgcg gtagccgctg
     2401 cgccctgtca atttcccttc cttattagcc gcttacggaa tgttcttaaa acattcactt
     2461 ttgcttatgt tttcgctgat atcccgagcg gtttcaaaat tgtgatctat atttaacaaa
     2521 gtgatgacat ttctgacggc gttaaatacc gttcaatgcg tagatatcag tatctaaagc
     2581 cgtcgattgt cattctaccg atattaataa ctgattcaga ggctgtaatg gtcgttattc
     2641 atcactcatc gcttttgtga tggcgaccat tgacttctgt agagggtgaa gtctctccct
     2701 attcagcaat gcaacctcgt gttgccaggc tcaaattacg agcaaacata caggaataaa
     2761 tcgatggcaa gcagcgaaga tgtgatcaaa gaatttatgc gtttcaaggt gcgtatggaa
     2821 ggtagcgtta atggtcatga atttgaaatc gaaggtgaag gtgagggtcg tccgtatgaa
     2881 ggcacccaga ccgccaaact gaaagttacc aaaggcggtc cgctgccgtt tgcatgggat
     2941 attctgagtc cgcagtttca gtatggtagc aaagcatatg ttaaacatcc ggcagatatc
     3001 ccggattatc tgaaactgag ctttccggaa ggctttaaat gggaacgtgt gatgaatttt
     3061 gaggatggtg gtgttgttac cgttacacag gatagcagcc tgcaggatgg tgaatttatc
     3121 tataaagtta aactgcgtgg cacgaatttt ccgtcagatg gtcctgttat gcagaaaaaa
     3181 accatgggtt gggaagcaag caccgaacgt atgtatccgg aagatggtgc actgaaaggt
     3241 gaaatcaaaa tgcgtctgaa gctgaaagat ggcggtcatt atgatgcaga agttaaaacc
     3301 acctacatgg ccaaaaaacc ggtgcagctg cctggtgcat ataaaacgga tattaaactg
     3361 gatatcacct cgcacaacga ggattatacc attgttgaac agtatgaacg tgcagaaggt
     3421 cgtcatagta ccggtgccta ataatactag agccaggcat caaataaaac gaaaggctca
     3481 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tctactagag
     3541 tcacactggc tcaccttcgg gtgggccttt ctgcgtttat aaaccgtgcg aaaagcctct
     3601 ttcggtgtta gctgggcctc atgggccttc ctttcactgc ccgctttcca gtcgggaaac
     3661 ctgtcgtgcc agctgcatta acatggtcat agctgtttcc ttgcgtattg ggcgctctcc
     3721 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggg taaagcctgg ggtgcctaat
     3781 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3841 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3901 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3961 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4021 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4081 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4141 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4201 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4261 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4321 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4381 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4441 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4501 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4561 tctaaagtat atatgagtaa acttggtctg acagttatta gaaaaattca tccagcagac
     4621 gataaaacgc aatacgctgg ctatccggtg ccgcaatgcc atacagcacc agaaaacgat
     4681 ccgcccattc gccgcccagt tcttccgcaa tatcacgggt ggccagcgca atatcctgat
     4741 aacgatccgc cacgcccaga cggccgcaat caataaagcc gctaaaacgg ccattttcca
     4801 ccataatgtt cggcaggcac gcatcaccat gggtcaccac cagatcttcg ccatccggca
     4861 tgctcgcttt cagacgcgca aacagctctg ccggtgccag gccctgatgt tcttcatcca
     4921 gatcatcctg atccaccagg cccgcttcca tacgggtacg cgcacgttca atacgatgtt
     4981 tcgcctgatg atcaaacgga caggtcgccg ggtccagggt atgcagacga cgcatggcat
     5041 ccgccataat gctcactttt tctgccggcg ccagatggct agacagcaga tcctgacccg
     5101 gcacttcgcc cagcagcagc caatcacggc ccgcttcggt caccacatcc agcaccgccg
     5161 cacacggaac accggtggtg gccagccagc tcagacgcgc cgcttcatcc tgcagctcgt
     5221 tcagcgcacc gctcagatcg gttttcacaa acagcaccgg acgaccctgc gcgctcagac
     5281 gaaacaccgc cgcatcagag cagccaatgg tctgctgcgc ccaatcatag ccaaacagac
     5341 gttccaccca cgctgccggg ctacccgcat gcaggccatc ctgttcaatc atactcttcc
     5401 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5461 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
//
