LOCUS       sequence-205146-        9035 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C. difficile inducible expression system. Shuttle plasmid containing
            a tetracycline-inducible gusA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9035)
  AUTHORS   Fagan RP, Fairweather NF
  TITLE     Clostridium difficile has two parallel and essential Sec secretion 
            systems.
  JOURNAL   J Biol Chem. 2011 Aug 5;286(31):27483-93. doi: 
            10.1074/jbc.M111.263889. Epub 2011 Jun 9.
  PUBMED    21659510
REFERENCE   2  (bases 1 to 9035)
  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"; doi: 
            "10.1074/jbc.M111.263889"; journalName: "J Biol Chem"; date: 
            "2011-08-5- 5"; volume: "286"; issue: "31"; pages: "27483-93"
COMMENT     Sequence Label: pRPF185
FEATURES             Location/Qualifiers
     source          1..9035
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(3225..3851)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     3756..3775
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     protein_bind    3958..3976
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     CDS             4043..5854
                     /codon_start=1
                     /gene="uidA (or gusA) from E. coli"
                     /product="beta-glucuronidase"
                     /label=GUS
                     /translation="MLRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     CDS             complement(6221..6592)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            6625..6734
                     /note="incP origin of transfer"
     primer_bind     complement(7925..7942)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8096..8684)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8176..8195)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctagctctat attctagtta tacattctct atattctagc tctatattct agttatacat
       61 tctctatatt ctagctctat attctagtta tacattctct atattctagc tctatattct
      121 agttatacat tctctatatt ctagctctat attctagtta tacattctct atattctagc
      181 tctatattct agttatacat tctctatatt ctagctctat attctagtta tacattctct
      241 aaaaaacata tatatatata ctacatatat atgtacaatt attttaccaa aagaaatttt
      301 taaaacggct taatttcaat actttgatgc catttttttt ttgcaattct cgtacaatta
      361 ttttaccaaa agaaattttt attaattagt ttttttgcat aaaaatagga ctagaaaatc
      421 aattctaatc ctatcaaaaa tctacttgtt taatctatat gtatagtaca taaatcctct
      481 atcatttatt ttagtttccc atacatctaa aattttagta tctaataaag attctatcat
      541 atttgaaatt ctttgctgtg ttactcctaa atcttttgct aattcctctt gattaattct
      601 taatatattt ccttttattt tatcatcttt gaccaataaa tcttgcttat atctcatatg
      661 agtatataat cttaattctt cagttgaaat tattccccaa attactgcca tggtaacaaa
      721 gtaactaata gtatattgtt tatctatatt acatctaact tcttttacat tgtatgtatc
      781 ttttattcct agctttgtaa caccttttgt ttttgtaata taatcattat gttgaagttc
      841 ttttaatgtt tctcttaatg tcttttcact cataacaatg tttttaactt tcttcttacg
      901 tttataggtt atcagctcca ttatatcgtc tatgtttaat tctctatctt tattgttaag
      961 taacacatta tagataaaca attgattacc agtcattatt ttaacccctt tcctattctt
     1021 atatttcaaa tcctttgagt gcttatgtgg catattgaac aaagtatctt catcactata
     1081 tataaaatct gattctattt tattccaaca atcactatta aactcacaac ctgtacaacc
     1141 aaaattaacc gttttaagct tctcatacat atacttgact tgatattcta gctcactatt
     1201 tgaaagacta ttttgtgata tatagttcca ttctctagca acggatatta cttgacttaa
     1261 acttttattc ctcaatctta aagtcacaac tattttttgc aaatctaggt ttttatgtcc
     1321 atctttacta ccatttttta aaatatctac tatacaattt ggtagcttac tatctaatat
     1381 atattttgta ttagtattac ctactgtttt tacatctttg caatattttt tagctaactt
     1441 ttctatgtca tatctatata tatttttgtc ttggtgtatt atttttactt gtttagtagt
     1501 atttttaata ttatatgtat aaggtactct taatacttga gtagataaat ttgcttgttt
     1561 atctgcaccc aatttattta ttaatgtatt tgtaacttct gatactaact taatgttatt
     1621 agttttatta attagcacat aaacatggaa tccatttcca ctatcaacta ggatatgata
     1681 atgtaatcct atcttagtaa ataaattaat tatatcttta taattaaaac cttgtcctaa
     1741 ttcttttttg tcaaaatcaa atcctaatgc atatctattt attaagtttt cattggttcc
     1801 tgcttttcca tcagttgtac ataggttaaa aaatgtattt tcataataaa tattttttgc
     1861 gaaggtaaat tctactatat catctatatt tttaaaaaaa gttgttttac tatattcttt
     1921 aaattcttct ccgagttcta attgtttact attactttta ttattttgaa atactctaac
     1981 atattcatta tcaataagat tatttccttg tcctatacca tctctaaata ctgccattag
     2041 aaattttttc aacttatatt ttgaatctaa ttgttccata tatttattat ctccttatct
     2101 agttttttta agagataaca ctacattaaa tatatactta ttgactaatc aacttttaca
     2161 actattgaat aattttagct tagtatgtat aataatattg ttaagatgct aaaattgata
     2221 gcaatattat tatataacta cagatactta atcagttaga ccgatattgg agttggtagc
     2281 tccataaact caattggtag ttgagttgtc attgtctagt agtattatct cattttgttg
     2341 ttatcttaaa atattatttt aaaaatactc tgcttggtcg gcttagggta ttttttttgt
     2401 ttaattccaa tattggttca aactcctgtg taatactttt gacacttcca tctgtattga
     2461 attgacagtt gatatggtat ctcattattg gcttgttctt atgtttttct atgtatttat
     2521 tttttacttt caaataattt gtatttccat ttttagttat gtttaacaca ctatccttat
     2581 tactcaataa aattattttt tgttttaaca tatgatttat agaatctaaa aattgatatt
     2641 gacttataat agtactttct acctctctag ctttttctag tgtttcaaat tcttcaaact
     2701 ctgctatatt gctatagtat agcttatcat taataatatt atctgacaac tttaattcca
     2761 ttttataaaa caattccaag ttatgtataa aatttttaat gttacaacat atagacttat
     2821 tatttgttct tatatggtat ctccatttac tacttctagc tctcatgtta tatgcacaac
     2881 tactcccaac ataaacatat attgcttgtt ctaactcata tttttgtaaa atttcaacat
     2941 catatccagt caaatcgtac tctctcttat atacaaatgg tttttctaat tttttgttca
     3001 ataaatctcc tcctatttct tacttaattt agctaaaaat ctatttatat ttcacctcct
     3061 ttttgacttt aagcctacga ataccatatt tatgattata acatcttcta tttttcccaa
     3121 atccttacat ctcccctaat ttttagactt aagggcgaat tctgcatcaa gctagcataa
     3181 aaataagaag cctgcatttg caggcttctt atttttatgg taccttaaga cccactttca
     3241 catttaagtt gtttttctaa tccgcatatg atcaattcaa ggccgaataa gaaggctggc
     3301 tctgcacctt ggtgatcaaa taattcgata gcttgtcgta ataatggcgg catactatca
     3361 gtagtaggtg tttccctttc ttctttagcg acttgatgct cttgatcttc caatacgcaa
     3421 cctaaagtaa aatgccccac agcgctgagt gcatataatg cattctctag tgaaaaacct
     3481 tgttggcata aaaaggctaa ttgattttcg agagtttcat actgtttttc tgtaggccgt
     3541 gtacctaaat gtacttttgc tccatcgcga tgacttagta aagcacatct aaaactttta
     3601 gcgttattac gtaaaaaatc ttgccagctt tccccttcta aagggcaaaa gtgagtatgg
     3661 tgcctatcta acatctcaat ggctaaggcg tcgagcaaag cccgcttatt ttttacatgc
     3721 caatacaatg taggctgctc tacacctagc ttctgggcga gtttacgggt tgttaaacct
     3781 tcgattccga cctcattaag cagctctaat gcgctgttaa tcactttact tttatctaat
     3841 ctagacatca ttaattcctc ctttttgttg acattatatc attgatagag ttatttgtca
     3901 aactagtttt ttatttcgat gccctggact tcatgaaaaa ctaaaaaaaa tattgacact
     3961 ctatcattga tagagtataa ttaaaataag cttgatcgta gcgttaacag atctgagctc
     4021 ctgcagtaaa ggagaaaatt ttatgttacg tcctgtagaa accccaaccc gtgaaatcaa
     4081 aaaactcgac ggcctgtggg cattcagtct ggatcgcgaa aactgtggaa ttgatcagcg
     4141 ttggtgggaa agcgcgttac aagaaagccg ggcaattgct gtgccaggca gttttaacga
     4201 tcagttcgcc gatgcagata ttcgtaatta tgcgggcaac gtctggtatc agcgcgaagt
     4261 ctttataccg aaaggttggg caggccagcg tatcgtgctg cgtttcgatg cggtcactca
     4321 ttacggcaaa gtgtgggtca ataatcagga agtgatggag catcagggcg gctatacgcc
     4381 atttgaagcc gatgtcacgc cgtatgttat tgccgggaaa agtgtacgta tcaccgtttg
     4441 tgtgaacaac gaactgaact ggcagactat cccgccggga atggtgatta ccgacgaaaa
     4501 cggcaagaaa aagcagtctt acttccatga tttctttaac tatgccggga tacatcgcag
     4561 cgtaatgctc tacaccacgc cgaacacctg ggtggacgat atcaccgtgg tgacgcatgt
     4621 cgcgcaagac tgtaaccacg cgtctgttga ctggcaggtg gtggccaatg gtgatgtcag
     4681 cgttgaactg cgtgatgcgg atcaacaggt ggttgcaact ggacaaggca ctagcgggac
     4741 tttgcaagtg gtgaatccgc acctctggca accgggtgaa ggttatctct atgaactgtg
     4801 cgtcacagcc aaaagccaga cagagtgtga tatctacccg cttcgcgtcg gcatccggtc
     4861 agtggcagtg aagggcgaac agttcctgat taaccacaaa ccgttctact ttactggctt
     4921 tggtcgtcat gaagatgcgg acttgcgtgg caaaggattc gataacgtgc tgatggtgca
     4981 cgaccacgca ttaatggact ggattggggc caactcctac cgtacctcgc attaccctta
     5041 cgctgaagag atgctcgact gggcagatga acatggcatc gtggtgattg atgaaactgc
     5101 tgctgtcggc tttaacctct ctttaggcat tggtttcgaa gcgggcaaca agccgaaaga
     5161 actgtacagc gaagaggcag tcaacgggga aactcagcaa gcgcacttac aggcgattaa
     5221 agagctgata gcgcgtgaca aaaaccaccc aagcgtggtg atgtggagta ttgccaacga
     5281 accggatacc cgtccgcaag gtgcacggga atatttcgcg ccactggcgg aagcaacgcg
     5341 taaactcgac ccgacgcgtc cgatcacctg cgtcaatgta atgttctgcg acgctcacac
     5401 cgataccatc agcgatctct ttgatgtgct gtgcctgaac cgttattacg gatggtatgt
     5461 ccaaagcggc gatttggaaa cggcagagaa ggtactggaa aaagaacttc tggcctggca
     5521 ggagaaactg catcagccga ttatcatcac cgaatacggc gtggatacgt tagccgggct
     5581 gcactcaatg tacaccgaca tgtggagtga agagtatcag tgtgcatggc tggatatgta
     5641 tcaccgcgtc tttgatcgcg tcagcgccgt cgtcggtgaa caggtatgga atttcgccga
     5701 ttttgcgacc tcgcaaggca tattgcgcgt tggcggtaac aagaaaggga tcttcactcg
     5761 cgaccgcaaa ccgaagtcgg cggcttttct gctgcaaaaa cgctggactg gcatgaactt
     5821 cggtgaaaaa ccgcagcagg gaggcaaaca atgaatcaac aactctcctg gcgcaccgga
     5881 tcctataagt tttaataaaa ctttaaatag aaaaaggctt ctctcatgag aagtcttttt
     5941 tatttaaaat aaatataaaa taaaatagag gctataaata gcctctattt tatgtgagaa
     6001 atccctaaat aaaaagatgc cagtgtgctg gaattcgccc ttaggctgcg ctagggacct
     6061 ctttagctcc ttggaagctg tcagtagtag gggatcggtc ttgccttgct cgtcggtgat
     6121 gtacttcacc agctccgcga agtcgctctt cttgatggag cgcatgggga cgtgcttggc
     6181 aatcacgcgc accccccggc cgttttagcg gctaaaaaag tcatggctct gccctcgggc
     6241 ggaccacgcc catcatgacc ttgccaagct cgtcctgctt ctcttcgatc ttcgccagca
     6301 gggcgaggat cgtggcatca ccgaaccgcg ccgtgcgcgg gtcgtcggtg agccagagtt
     6361 tcagcaggcc gcccaggcgg cccaggtcgc cattgatgcg ggccagctcg cggacgtgct
     6421 catagtccac gacgcccgtg attttgtagc cctggccgac ggccagcagg taggccgaca
     6481 ggctcatgcc ggccgccgcc gccttttcct caatcgctct tcgttcgtct ggaaggcagt
     6541 acaccttgat aggtgggctg cccttcctgg ttggcttggt ttcatcagcc atccgcttgc
     6601 cctcatctgt tacgccggcg gtagccggcc agcctcgcag agcaggattc ccgttgagca
     6661 ccgccaggtg cgaataaggg acagtgaaga aggaacaccc gctcgcgggt gggcctactt
     6721 cacctatcct gcccggctga cgccgttgga tacaccaagg aaagtctaca cgaacccttt
     6781 ggcaaaatcc tgtatatcgt gcgaaaaagg atggatatac cgaaaaaatc gctataatga
     6841 ccccgaagca ggaagggcga attctgcaga tctgaccggt ctctgaaaat ataaaaacca
     6901 cagattgata ctaaaacctt ggttgtgttg cttttcgggg cttaaatcaa ggaaaaatcc
     6961 ttgttttaag cctttcaaaa agaaacacaa ggtctttgta ctaacctgtg gttatgtata
     7021 aaattgtaga ttttagggta acaaaaaaca ccgtatttct acgatgtttt tgcttaaata
     7081 cttgttttta gttacagaca aacctgaagt taactattta tcaattcctg caattcgttt
     7141 acaaaacggc aaatgtgaaa tccgtcacat actgcgtgat gaacttgaat tgccaaagga
     7201 agtataattt tgttatcttc tttataatat ttccccatag taaaaatagg aatcaaataa
     7261 tcatatcctt tctgcaaatt cagattaaag ccatcgaagg ttgaccacgg tatcatagat
     7321 acattaaaaa tgttttccgg agcatttggc tttccttcca ttctatgatt gtttccatac
     7381 cgttgcgtat cactttcata atctgctaaa aatgatttaa agtcagactt acactcagtc
     7441 caaaggctgg aaaatgtttc agtatcattg tgaaatattg tatagcttgg tatcatctca
     7501 tcatatatcc ccaattcacc atcttgattg attgccgtcc taaactctga atggcggttt
     7561 acaatcattg caatataata aagcattgca ggatatagtt tcattccctt ttcctttatt
     7621 tgtgtgatat ccactttaac ggtcatgctg taggtacaag gtacacttgc aaagtagtgg
     7681 tcaaaatact cttttctgtt ccaactattt ttatcaattt tttcaaatac catctaagtt
     7741 ccctctcaaa ttcaagttta tcgctctaat gaacaaagat attataccac atttttgtga
     7801 atttttcaac ttgcccactt cgactgcact cccgacttaa taacttcttg aacacttgcc
     7861 gaaaaagaaa aactgccggg tacgtacccg gggatcgatc cccggccgag cgctcttccg
     7921 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     7981 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     8041 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     8101 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     8161 acccgacagg actataaaga taccaggcgt ttcccccttg aagctccctc gtgcgctctc
     8221 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     8281 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     8341 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     8401 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     8461 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     8521 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     8581 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     8641 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     8701 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     8761 aatgcaagtt tctaactaac atacatcata gttactaaac tatggtaact ataattttat
     8821 taactatatt aagcactgat tagtactata actcaatata agcatatccc ctgtatcgta
     8881 actagagaac caaacgacgg aaaaagcgat atagataagt ttagatagtg caccaatttt
     8941 ttacttttct attttagaat ctagcatttc caatgcttac taatactatt tttgattttt
     9001 aacttgtttt aaaaaagcac ctactaatta ggatt
//
