LOCUS       sequence-178321-        5365 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ThsS under the Ptac promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5365)
  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 5365)
  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: pKD182
FEATURES             Location/Qualifiers
     source          1..5365
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             149..940
                     /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      952..1046
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      1160..1705
                     /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     1606..1625
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1822..1839
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        1878..1955
                     /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             1956..3038
                     /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(1975..1994)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     terminator      3044..3130
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        join(3303..3308,3309..3324,3325..3331)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    3339..3355
                     /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)."
     terminator      5200..5271
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5287..5314
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 agcaagcgaa ccggaattgc cagctggggc gccctctggt aaggttggga agccctgcaa
       61 agtaaactgg atggctttct tgccgccaag gatctgatgg cgcaggggat caagatctga
      121 tcaagagaca ggatgaggat cgtttcgtat gagggaagcg gtgatcgccg aagtatcgac
      181 tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt
      241 acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagtgata ttgatttgct
      301 ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca acgacctttt
      361 ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag tcaccattgt
      421 tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc aatttggaga
      481 atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg acattgatct
      541 ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc cagcggcgga
      601 ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg aaaccttaac
      661 gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc ttacgttgtc
      721 ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg
      781 ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta gacaggctta
      841 tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag aatttgtcca
      901 ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa gagctcgctt ggactcctgt
      961 tgatagatcc agtaatgacc tcagaactcc atctggattt gttcagaacg ctcggttgcc
     1021 gccgggcgtt ttttattggt gagaatccaa gcactagtaa caacttatat cgtatggggc
     1081 tgacttcagg tgctacattt gaagagataa attgcactga aatctagtaa tattttatct
     1141 gattaataag atgatcttct tgagatcgtt ttggtctgcg cgtaatctct tgctctgaaa
     1201 acgaaaaaac cgccttgcag ggcggttttt cgaaggttct ctgagctacc aactctttga
     1261 accgaggtaa ctggcttgga ggagcgcagt caccaaaact tgtcctttca gtttagcctt
     1321 aaccggcgca tgacttcaag actaactcct ctaaatcaat taccagtggc tgctgccagt
     1381 ggtgcttttg catgtctttc cgggttggac tcaagacgat agttaccgga taaggcgcag
     1441 cggtcggact gaacgggggg ttcgtgcata cagtccagct tggagcgaac tgcctacccg
     1501 gaactgagtg tcaggcgtgg aatgagacaa acgcggccat aacagcggaa tgacaccggt
     1561 aaaccgaaag gcaggaacag gagagcgcac gagggagccg ccagggggaa acgcctggta
     1621 tctttatagt cctgtcgggt ttcgccacca ctgatttgag cgtcagattt cgtgatgctt
     1681 gtcagggggg cggagcctat ggaaaaacgg ctttgccgcg gccctctcac ttccctgtta
     1741 agtatcttcc tggcatcttc cgggaaatct ccgccccgtt cgtaagccat ttccgctcgc
     1801 cgcagtcgaa cgaccgagcg tagcgagtca gtgagcgagg aagcggaata tatcccgaag
     1861 cggcatgcat ttacgttgac accatcgaat ggtgcaaaac ctttcgcggt atggcatgat
     1921 agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg
     1981 tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc
     2041 acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg aattacattc
     2101 ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc gttgccacct
     2161 ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc
     2221 aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag
     2281 cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac tatccgctgg
     2341 atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg ttatttcttg
     2401 atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac ggtacgcgac
     2461 tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta gcgggcccat
     2521 taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc actcgcaatc
     2581 aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt tttcaacaaa
     2641 ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc aacgatcaga
     2701 tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt gcggatatct
     2761 cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg ttaaccacca
     2821 tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg caactctctc
     2881 agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa agaaaaacca
     2941 ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     3001 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagg catcaaataa aacgaaaggc
     3061 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
     3121 taggacaaat ccgccgccct agacctaggg cgttcggctg cggcgagcgg tatcagctca
     3181 ctcaaaggcg gtaatacgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
     3241 tctggataat gttttttgcg ccgacatcat aacggttctg gctaatattc tgaaatgagc
     3301 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
     3361 cacaaacgac catcgacgtc cgaccctaac gtatactatg tcccgcctgc tgctgtgtat
     3421 ctgtgttctg ctgttctctt ctgtggcgtg gtctaaaccg cagcagtttt atgtgggcgt
     3481 actggctaac tggggtcatc agcaagccgt tgaacgttgg accccgatga tggagtatct
     3541 gaacgaacat gtgccggacg cggaatttca cgtctacccg ggcaacttca aagcactgaa
     3601 cctggcaatg gaactgggcc agattcagtt cattatcact aacccgggcc aatatctgta
     3661 cctgagcaat cagtacccgc tgtcttggct ggcgaccatg cgttctaagc gtcacgatgg
     3721 taccacttct gcgatcggtt ccgccattat tgtccgcgcg gacagcgact accgcaccct
     3781 gtacgacctg aaaggtaaag tggtggctgc gtccgacccg catgctctgg gtggctacca
     3841 agcgaccgtc ggtctgatgc attccctggg catggatccg gacaccttct tcggtgaaac
     3901 caagtttctg ggctttccac tggatccgct gctgtaccaa gttcgtgatg gcaacgttga
     3961 cgcggccatt accccactgt gcactctgga ggacatggtt gcacgcggcg tactgaaatc
     4021 ttccgatttt cgtgtgctga accctagccg cccggatggt gtagaatgcc agtgctctac
     4081 caccctgtac ccgaactggt ctttcgctgc gactgagtct gtatccaccg aactgtctaa
     4141 agaaatcacg caggcactgc tggaactgcc atccgacagc ccggcagcta tcaaagcgca
     4201 actgaccggc tggaccagcc cgatctccca actggcggta atcaaactgt tcaaagagct
     4261 gcacgtaaaa accccggact ctagccgttg ggaagccgtt aagaagtggc tggaagaaaa
     4321 ccgtcactgg ggtatcctgt ctgttctggt gttcatcatt gcaacgctgt atcacctgtg
     4381 gattgaatac cgcttccacc aaaaaagctc ttctctgatc gaatctgaac gtcagctgaa
     4441 acagcaagct gttgccctgg aacgtctgca atctgctagc atcgttggtg aaattggtgc
     4501 gggtctggcc cacgagatta atcagccgat cgctgcaatt acctcttatt ctgaaggtgg
     4561 catcatgcgc ctgcaaggta aagaacaggc ggatacggat agctgcatcg aactgctgga
     4621 aaaaatccac aaacagagca ctcgcgcagg cgaagtggtg caccgcatcc gtggtctgct
     4681 gaaacgtcgt gaagcggtga tggtagatgt taacatcctg accctggtgg aagaatccat
     4741 cagcctgctg cgtctggagc tggcacgtcg cgaaatccag atcaacactc agatcaaagg
     4801 tgaaccgttc ttcattactg ccgaccgcgt tggcctgctg caagttctga ttaacctgat
     4861 caaaaactcc ctggacgcga tcgctgaatc tgataatgcc cgttctggta aaatcaacat
     4921 cgaactggac tttaaagagt accaggtaaa cgtctccatc atcgataacg gtccgggcct
     4981 ggcgatggat tctgacactc tgatggctac gttttacact accaaaatgg atggcctggg
     5041 cctgggtctg gcaatctgcc gcgaagttat cagcaaccac gacggccact tcctgctgtc
     5101 caaccgtgac gacggcgttc tgggctgtgt ggcaaccctg aatctgaaaa aacgcggttc
     5161 tgaagtgccg atcgaagtct aataatctag accaggcatc aaataaaacg aaaggctcag
     5221 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt
     5281 cacactggct caccttcggg tgggcctttc tgcgtttata ggtacccggg gatcctctag
     5341 agtcgacctg caggcatgca agctt
//
