LOCUS       sequence-178329-        4557 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses S. baltica TtrR and mCherry constitutively and sfGFP under
            the PttrB185-269 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1711 to 1745)
  AUTHORS   Daeffler KN, Galley JD, Sheth RU, Ortiz-Velez LC, Bibb CO, Shroyer 
            NF, Britton RA, Tabor JJ
  TITLE     Engineering bacterial thiosulfate and tetrathionate sensors for 
            detecting gut inflammation.
  JOURNAL   Mol Syst Biol. 2017 Apr 3;13(4):923. doi: 10.15252/msb.20167416.
  PUBMED    28373240
REFERENCE   2  (bases 1 to 4557)
  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: "Mol Syst 
            Biol."; date: "2017-04-3"; volume: "13(4):923. doi"; pages: " 
            10.15252/msb.20167416"
COMMENT     Sequence Label: pKD239-1g-2
FEATURES             Location/Qualifiers
     source          1..4557
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(132..720)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(212..231)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(808..902)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(923..1582)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVTCYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1497..1516
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        1711..1745
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     RBS             1755..1766
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1774..2484
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1919..1937)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2187..2206
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2585..2603
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2624..2658
                     /label=BBa_J23109
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.04"
     terminator      3329..3400
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3416..3443
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             3652..3663
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3670..4383
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHMLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      4410..4481
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4497..4524
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
       61 gcatgcaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
      121 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
      181 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
      241 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
      301 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
      361 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
      421 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
      481 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
      541 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
      601 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
      661 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
      721 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
      781 gggattttgg tcatgactag tgcttggatt ctcaccaata aaaaacgccc ggcggcaacc
      841 gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag
      901 tccaagcgag ctcgatatca aattacgccc cgccctgcca ctcatcgcag tactgttgta
      961 attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg
     1021 ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatggtg aaaacggggg
     1081 cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat
     1141 tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac
     1201 cataacacgt cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt
     1261 cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa
     1321 cactatccca tatcaccagc tcaccgtctt tcattgccat acgaaattcc ggatgagcat
     1381 tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta
     1441 cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa
     1501 ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat
     1561 atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc gataactcaa
     1621 aaaatacgac cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgggcc
     1681 ggccaacgtc tcattttcgc cagatatcga ttgacggcta gctcagtcct aggtacagtg
     1741 ctagcgaatt cattaaagag gagaaaggta cccatggtga gcaagggcga ggaggataac
     1801 atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc
     1861 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     1921 aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag
     1981 ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag
     2041 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg
     2101 gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg
     2161 cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     2221 gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg
     2281 ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag
     2341 aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac
     2401 aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc
     2461 ggcatggacg agctgtacaa gtaagctttc taactaaaaa ggcctcccaa atcggggggc
     2521 cttttttatt gataacaaaa cgctcgtcta agaaaccatt attatcatga cattaaccta
     2581 taaaaatagg cgtatcacga ggccctttcg tcttcacctc gagtttacag ctagctcagt
     2641 cctagggact gtgctagcac caaatataga gaatcacaag gatctattca atgagcctgg
     2701 cgctgccggt gtacctgatt gacgacgatg actccgttcg ccgttccctg cgtttcatgc
     2761 tggaaagcta cggcctgaaa atcactgact tcgattctgc tgaagcgttc ttcaccgcgg
     2821 tagacctgac cctgccgggt tgcgcactgg tggacgtacg tatgccgggc ctgagcggcc
     2881 cacagctgca cctggaactg gtgtctaaga acagcccgct ggccgtgatc tatctgaccg
     2941 gtcacggcga cgttccgatg gcggttgaag cgctgaaact gggtgcggta gatttctttc
     3001 agaaacctgc agacggcgct aaactggctg aagctgtggt caaagccctg gaacacgcga
     3061 aaacccacta ccaagacaac cagtacctgg aaacctatca ggctctgacc ccacgtgagc
     3121 gcgaaatcct gaacctgatt gcgcaaggtc tgaaaaacca ggaaatcgcg gacaacctgt
     3181 gcattgcgat gcgcaccgta gaaatccacc gtgcgaacct gatgaaaggt atgcaggtgg
     3241 gcagcctggc agagatgatg ctgatttact ctcgtatcgc ggagcgtctg cgcctggaag
     3301 ataaccgtta ataatctaga ccaggcatca aataaaacga aaggctcagt cgaaagactg
     3361 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc tactagagtc acactggctc
     3421 accttcgggt gggcctttct gcgtttatag taaatcactg cataattcgt gtcgctcaag
     3481 gcgcactccc gttctggata atgttttttg cgccgacatc ataacggttc tggcaaatat
     3541 tctgaaatga gctggttagc tagtcaatat ttgttgcgct agatcaaatc cacgcctgat
     3601 atgtggaaaa ccactatagt tatgccgcct cgccttttac agaatgcctg taaagaggag
     3661 aaatactaga tgcgtaaagg cgaagagctg ttcactggtg tcgtccctat tctggtggaa
     3721 ctggatggtg atgtcaacgg tcataagttt tccgtgcgtg gcgagggtga aggtgacgca
     3781 actaatggta aactgacgct gaagttcatc tgtactactg gtaaactgcc ggtaccttgg
     3841 ccgactctgg taacgacgct gacttatggt gttcagtgct ttgctcgtta tccggaccat
     3901 atgaagcagc atgacttctt caagtccgcc atgccggaag gctatgtgca ggaacgcacg
     3961 atttccttta aggatgacgg cacgtacaaa acgcgtgcgg aagtgaaatt tgaaggcgat
     4021 accctggtaa accgcattga gctgaaaggc attgacttta aagaagacgg caatatcctg
     4081 ggccatatgc tggaatacaa ttttaacagc cacaatgttt acatcaccgc cgataaacaa
     4141 aaaaatggca ttaaagcgaa ttttaaaatt cgccacaacg tggaggatgg cagcgtgcag
     4201 ctggctgatc actaccagca aaacactcca atcggtgatg gtcctgttct gctgccagac
     4261 aatcactatc tgagcacgca aagcgttctg tctaaagatc cgaacgagaa acgcgatcat
     4321 atggttctgc tggagttcgt aaccgcagcg ggcatcacgc atggtatgga tgaactgtac
     4381 aaatgatgat aataatctag accaggcatc aaataaaacg aaaggctcag tcgaaagact
     4441 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct
     4501 caccttcggg tgggcctttc tgcgtttata ggtacccgtg gatcctctag agtcgac
//
