LOCUS       sequence-178325-        5597 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses S. baltica TtrS under the Ptac promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5597)
  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 5597)
  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: pKD227
FEATURES             Location/Qualifiers
     source          1..5597
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     163..180
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        219..296
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             297..1379
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(316..335)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     terminator      1385..1471
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        join(1644..1649,1650..1665,1666..1672)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1680..1696
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     regulatory      2832..2841
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     terminator      3773..3844
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3860..3887
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             4087..4878
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     terminator      4890..4984
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      join(5098..5597,1..46)
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     5544..5563
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgtcagatt tcgtgatgct tgtcaggggg gcggagccta tggaaaaacg gctttgccgc
       61 ggccctctca cttccctgtt aagtatcttc ctggcatctt ccgggaaatc tccgccccgt
      121 tcgtaagcca tttccgctcg ccgcagtcga acgaccgagc gtagcgagtc agtgagcgag
      181 gaagcggaat atatcccgaa gcggcatgca tttacgttga caccatcgaa tggtgcaaaa
      241 cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga
      301 aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc
      361 gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga
      421 tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt
      481 tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg
      541 cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa
      601 gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc
      661 tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta
      721 atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt attattttct
      781 cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa
      841 tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc
      901 ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg
      961 ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga
     1021 tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc
     1081 tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga taccgaagac agctcatgtt
     1141 atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg
     1201 accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct
     1261 cactggtgaa aagaaaaacc accctggcgc ccaatacgca aaccgcctct ccccgcgcgt
     1321 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     1381 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     1441 gtcggtgaac gctctcctga gtaggacaaa tccgccgccc tagacctagg gcgttcggct
     1501 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgt aaatcactgc ataattcgtg
     1561 tcgctcaagg cgcactcccg ttctggataa tgttttttgc gccgacatca taacggttct
     1621 ggctaatatt ctgaaatgag ctgttgacaa ttaatcatcg gctcgtataa tgtgtggaat
     1681 tgtgagcgga taacaatttc acacatatac ctcttacact actatataaa ttttcttcca
     1741 tgttttcttc cggcatgagc ctgaaatggc ctctgagcat ccagggttcc atcgaaccga
     1801 aaatgactcc gcgcatcatc ttccgcatgg ccctgcgcac taaagtgttc atcaccctgc
     1861 tggcttgcct gtttgcttct gccaacgtat tcgcggtgga accgcagaac cagtccgcgc
     1921 cttccgatgt gatggacgct gaggtggttg ctccggttgc gacggaggaa ccgggttacc
     1981 aggtcgttga tgttggtgtg ctggcgattc gcggttataa agctaccatc aaccgttggc
     2041 agccactgat gggttggctg gagactcaga tcccgaactc ctacttccgc ctgcacccgc
     2101 tgtctctgga cgagctggca aagggtgtag aaacccaggg tctggacttc gtaatcacca
     2161 atccaggcca gtccgtactg ctggcgcgtc agtattctct gacgtggctg gcgaccctgc
     2221 gctctccgct gaacaacggt gctgcgatgc aggtgggttc cgcgctggta gttcgtgccg
     2281 acagcccgta ccagaccctg accgatctga aaggccgtcg tctgggcatc gtcagcaaaa
     2341 acgcattcgg tggttatctg acgctggtat atgaggcaca gctgaaaggc atcgatctgc
     2401 cgcgctttgt tggtgaaatc atcccactgg gcttcccgct ggataacctg ctgtatcagc
     2461 tggatgatca gaaatctggt aatgacgccg caaaagacaa tcgtctggat gccaccgttg
     2521 taccggtatg ccaactggaa cagatgcagg ccgaaggtct gatcaacatc ggccactatc
     2581 gcgttctgga caaccagacc ccggtaggtt tccactgtca agtaagcact cgtctgtatc
     2641 cgaactggtc tatggctaaa actaaccgtg ccagccagtc cctggcaaaa tctgtgaccc
     2701 aggctctgct ggctctgccg gaagatcacc tggcagcgaa aagctctgac tctgcaggtt
     2761 ggaccactgc agtgtcccag ctggcgatcg accagctgct gaaagacctg gacatgcatc
     2821 cactgcagac cccatggtgg cagcgcgctt ggcagtgggt aaaactgcac cagcagtggg
     2881 cgtggttcat tctgggtatc ctggtcctgc tgaacgcata ccatttctgg ctggaatacc
     2941 gtttctcccg tcgtggtcgt gaactgatca acacccagcg ccagctgaac gaaaaccgtg
     3001 ctctgctgga gcacgcacaa cgtatcgcca tcgctggcga actgggtgct agcctgtccc
     3061 atgaactgaa ccagccgctg gctgcaattg gtcactattg ccatggtgcg gaagtgcgtc
     3121 tgcagcgtgg tacttctcct gaagaactgc agtctgttct gaccctgatc cagcaggaag
     3181 ttacgcgcgc cgactctatc atcagccgtc tgcgtaacct gctgaaaaaa cgcccggtta
     3241 gcaaacaggc tctgtatctg cacgaactgg taaacgaaac ggtgccgctg ctggcgtacg
     3301 aattcgaaca gcaccagatc aacctggcgg taaacgtcag cggtgaacct tacctgcaat
     3361 ctctggacga ggtcggtatg cagcagctgc tgctgaacct gctgaaaaac gcgtttgacg
     3421 cgtgcgttca gcgcctggaa ctggaatcct ccggcactga acagagcggt atgcgtaaac
     3481 cgtatgttcc tacgatcgat attgatctgc gctaccagga gcgtactctg ctgctgactg
     3541 tgacggataa cggcactggt ctgaccgagg agactagcct gctgatgcag gccttctatt
     3601 ccaccaaatc tgaaggcctg ggcctgggtc tggttatttg ccgtgatatc gctgagtccc
     3661 acggtggcac cttctctctg gaatccgcca tgggtggtgg ctgccaggct caggtggcca
     3721 tcccgcgcaa gccggaaccg aatggcgttc tgtaataatc tagaccaggc atcaaataaa
     3781 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     3841 tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgttt ataggtaccc
     3901 ggggatcctc tagagtcgac ctgcaggcat gcaagcttag caagcgaacc ggaattgcca
     3961 gctggggcgc cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttcttg
     4021 ccgccaagga tctgatggcg caggggatca agatctgatc aagagacagg atgaggatcg
     4081 tttcgtatga gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc
     4141 gtcatcgagc gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg
     4201 gatggcggcc tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt
     4261 gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga
     4321 gagagcgaga ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg
     4381 tggcgttatc cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt
     4441 gcaggtatct tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca
     4501 agagaacata gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct
     4561 gaacaggatc tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac
     4621 tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta
     4681 accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc
     4741 cagtatcagc ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc
     4801 ttggcctcgc gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc
     4861 aaggtagtcg gcaaataaga gctcgcttgg actcctgttg atagatccag taatgacctc
     4921 agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtga
     4981 gaatccaagc actagtaaca acttatatcg tatggggctg acttcaggtg ctacatttga
     5041 agagataaat tgcactgaaa tctagtaata ttttatctga ttaataagat gatcttcttg
     5101 agatcgtttt ggtctgcgcg taatctcttg ctctgaaaac gaaaaaaccg ccttgcaggg
     5161 cggtttttcg aaggttctct gagctaccaa ctctttgaac cgaggtaact ggcttggagg
     5221 agcgcagtca ccaaaacttg tcctttcagt ttagccttaa ccggcgcatg acttcaagac
     5281 taactcctct aaatcaatta ccagtggctg ctgccagtgg tgcttttgca tgtctttccg
     5341 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcggactga acggggggtt
     5401 cgtgcataca gtccagcttg gagcgaactg cctacccgga actgagtgtc aggcgtggaa
     5461 tgagacaaac gcggccataa cagcggaatg acaccggtaa accgaaaggc aggaacagga
     5521 gagcgcacga gggagccgcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5581 cgccaccact gatttga
//
