LOCUS       sequence-201386-        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(3333..3959)
                     /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     3864..3883
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     protein_bind    4066..4084
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     CDS             4151..5962
                     /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(6329..6700)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            6733..6842
                     /note="incP origin of transfer"
     primer_bind     complement(8033..8050)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8204..8792)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8284..8303)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtgcaccaat tttttacttt tctattttag aatctagcat ttccaatgct tactaatact
       61 atttttgatt tttaacttgt tttaaaaaag cacctactaa ttaggattct agctctatat
      121 tctagttata cattctctat attctagctc tatattctag ttatacattc tctatattct
      181 agctctatat tctagttata cattctctat attctagctc tatattctag ttatacattc
      241 tctatattct agctctatat tctagttata cattctctat attctagctc tatattctag
      301 ttatacattc tctatattct agctctatat tctagttata cattctctaa aaaacatata
      361 tatatatact acatatatat gtacaattat tttaccaaaa gaaattttta aaacggctta
      421 atttcaatac tttgatgcca tttttttttt gcaattctcg tacaattatt ttaccaaaag
      481 aaatttttat taattagttt ttttgcataa aaataggact agaaaatcaa ttctaatcct
      541 atcaaaaatc tacttgttta atctatatgt atagtacata aatcctctat catttatttt
      601 agtttcccat acatctaaaa ttttagtatc taataaagat tctatcatat ttgaaattct
      661 ttgctgtgtt actcctaaat cttttgctaa ttcctcttga ttaattctta atatatttcc
      721 ttttatttta tcatctttga ccaataaatc ttgcttatat ctcatatgag tatataatct
      781 taattcttca gttgaaatta ttccccaaat tactgccatg gtaacaaagt aactaatagt
      841 atattgttta tctatattac atctaacttc ttttacattg tatgtatctt ttattcctag
      901 ctttgtaaca ccttttgttt ttgtaatata atcattatgt tgaagttctt ttaatgtttc
      961 tcttaatgtc ttttcactca taacaatgtt tttaactttc ttcttacgtt tataggttat
     1021 cagctccatt atatcgtcta tgtttaattc tctatcttta ttgttaagta acacattata
     1081 gataaacaat tgattaccag tcattatttt aacccctttc ctattcttat atttcaaatc
     1141 ctttgagtgc ttatgtggca tattgaacaa agtatcttca tcactatata taaaatctga
     1201 ttctatttta ttccaacaat cactattaaa ctcacaacct gtacaaccaa aattaaccgt
     1261 tttaagcttc tcatacatat acttgacttg atattctagc tcactatttg aaagactatt
     1321 ttgtgatata tagttccatt ctctagcaac ggatattact tgacttaaac ttttattcct
     1381 caatcttaaa gtcacaacta ttttttgcaa atctaggttt ttatgtccat ctttactacc
     1441 attttttaaa atatctacta tacaatttgg tagcttacta tctaatatat attttgtatt
     1501 agtattacct actgttttta catctttgca atatttttta gctaactttt ctatgtcata
     1561 tctatatata tttttgtctt ggtgtattat ttttacttgt ttagtagtat ttttaatatt
     1621 atatgtataa ggtactctta atacttgagt agataaattt gcttgtttat ctgcacccaa
     1681 tttatttatt aatgtatttg taacttctga tactaactta atgttattag ttttattaat
     1741 tagcacataa acatggaatc catttccact atcaactagg atatgataat gtaatcctat
     1801 cttagtaaat aaattaatta tatctttata attaaaacct tgtcctaatt cttttttgtc
     1861 aaaatcaaat cctaatgcat atctatttat taagttttca ttggttcctg cttttccatc
     1921 agttgtacat aggttaaaaa atgtattttc ataataaata ttttttgcga aggtaaattc
     1981 tactatatca tctatatttt taaaaaaagt tgttttacta tattctttaa attcttctcc
     2041 gagttctaat tgtttactat tacttttatt attttgaaat actctaacat attcattatc
     2101 aataagatta tttccttgtc ctataccatc tctaaatact gccattagaa attttttcaa
     2161 cttatatttt gaatctaatt gttccatata tttattatct ccttatctag tttttttaag
     2221 agataacact acattaaata tatacttatt gactaatcaa cttttacaac tattgaataa
     2281 ttttagctta gtatgtataa taatattgtt aagatgctaa aattgatagc aatattatta
     2341 tataactaca gatacttaat cagttagacc gatattggag ttggtagctc cataaactca
     2401 attggtagtt gagttgtcat tgtctagtag tattatctca ttttgttgtt atcttaaaat
     2461 attattttaa aaatactctg cttggtcggc ttagggtatt ttttttgttt aattccaata
     2521 ttggttcaaa ctcctgtgta atacttttga cacttccatc tgtattgaat tgacagttga
     2581 tatggtatct cattattggc ttgttcttat gtttttctat gtatttattt tttactttca
     2641 aataatttgt atttccattt ttagttatgt ttaacacact atccttatta ctcaataaaa
     2701 ttattttttg ttttaacata tgatttatag aatctaaaaa ttgatattga cttataatag
     2761 tactttctac ctctctagct ttttctagtg tttcaaattc ttcaaactct gctatattgc
     2821 tatagtatag cttatcatta ataatattat ctgacaactt taattccatt ttataaaaca
     2881 attccaagtt atgtataaaa tttttaatgt tacaacatat agacttatta tttgttctta
     2941 tatggtatct ccatttacta cttctagctc tcatgttata tgcacaacta ctcccaacat
     3001 aaacatatat tgcttgttct aactcatatt tttgtaaaat ttcaacatca tatccagtca
     3061 aatcgtactc tctcttatat acaaatggtt tttctaattt tttgttcaat aaatctcctc
     3121 ctatttctta cttaatttag ctaaaaatct atttatattt cacctccttt ttgactttaa
     3181 gcctacgaat accatattta tgattataac atcttctatt tttcccaaat ccttacatct
     3241 cccctaattt ttagacttaa gggcgaattc tgcatcaagc tagcataaaa ataagaagcc
     3301 tgcatttgca ggcttcttat ttttatggta ccttaagacc cactttcaca tttaagttgt
     3361 ttttctaatc cgcatatgat caattcaagg ccgaataaga aggctggctc tgcaccttgg
     3421 tgatcaaata attcgatagc ttgtcgtaat aatggcggca tactatcagt agtaggtgtt
     3481 tccctttctt ctttagcgac ttgatgctct tgatcttcca atacgcaacc taaagtaaaa
     3541 tgccccacag cgctgagtgc atataatgca ttctctagtg aaaaaccttg ttggcataaa
     3601 aaggctaatt gattttcgag agtttcatac tgtttttctg taggccgtgt acctaaatgt
     3661 acttttgctc catcgcgatg acttagtaaa gcacatctaa aacttttagc gttattacgt
     3721 aaaaaatctt gccagctttc cccttctaaa gggcaaaagt gagtatggtg cctatctaac
     3781 atctcaatgg ctaaggcgtc gagcaaagcc cgcttatttt ttacatgcca atacaatgta
     3841 ggctgctcta cacctagctt ctgggcgagt ttacgggttg ttaaaccttc gattccgacc
     3901 tcattaagca gctctaatgc gctgttaatc actttacttt tatctaatct agacatcatt
     3961 aattcctcct ttttgttgac attatatcat tgatagagtt atttgtcaaa ctagtttttt
     4021 atttcgatgc cctggacttc atgaaaaact aaaaaaaata ttgacactct atcattgata
     4081 gagtataatt aaaataagct tgatcgtagc gttaacagat ctgagctcct gcagtaaagg
     4141 agaaaatttt atgttacgtc ctgtagaaac cccaacccgt gaaatcaaaa aactcgacgg
     4201 cctgtgggca ttcagtctgg atcgcgaaaa ctgtggaatt gatcagcgtt ggtgggaaag
     4261 cgcgttacaa gaaagccggg caattgctgt gccaggcagt tttaacgatc agttcgccga
     4321 tgcagatatt cgtaattatg cgggcaacgt ctggtatcag cgcgaagtct ttataccgaa
     4381 aggttgggca ggccagcgta tcgtgctgcg tttcgatgcg gtcactcatt acggcaaagt
     4441 gtgggtcaat aatcaggaag tgatggagca tcagggcggc tatacgccat ttgaagccga
     4501 tgtcacgccg tatgttattg ccgggaaaag tgtacgtatc accgtttgtg tgaacaacga
     4561 actgaactgg cagactatcc cgccgggaat ggtgattacc gacgaaaacg gcaagaaaaa
     4621 gcagtcttac ttccatgatt tctttaacta tgccgggata catcgcagcg taatgctcta
     4681 caccacgccg aacacctggg tggacgatat caccgtggtg acgcatgtcg cgcaagactg
     4741 taaccacgcg tctgttgact ggcaggtggt ggccaatggt gatgtcagcg ttgaactgcg
     4801 tgatgcggat caacaggtgg ttgcaactgg acaaggcact agcgggactt tgcaagtggt
     4861 gaatccgcac ctctggcaac cgggtgaagg ttatctctat gaactgtgcg tcacagccaa
     4921 aagccagaca gagtgtgata tctacccgct tcgcgtcggc atccggtcag tggcagtgaa
     4981 gggcgaacag ttcctgatta accacaaacc gttctacttt actggctttg gtcgtcatga
     5041 agatgcggac ttgcgtggca aaggattcga taacgtgctg atggtgcacg accacgcatt
     5101 aatggactgg attggggcca actcctaccg tacctcgcat tacccttacg ctgaagagat
     5161 gctcgactgg gcagatgaac atggcatcgt ggtgattgat gaaactgctg ctgtcggctt
     5221 taacctctct ttaggcattg gtttcgaagc gggcaacaag ccgaaagaac tgtacagcga
     5281 agaggcagtc aacggggaaa ctcagcaagc gcacttacag gcgattaaag agctgatagc
     5341 gcgtgacaaa aaccacccaa gcgtggtgat gtggagtatt gccaacgaac cggatacccg
     5401 tccgcaaggt gcacgggaat atttcgcgcc actggcggaa gcaacgcgta aactcgaccc
     5461 gacgcgtccg atcacctgcg tcaatgtaat gttctgcgac gctcacaccg ataccatcag
     5521 cgatctcttt gatgtgctgt gcctgaaccg ttattacgga tggtatgtcc aaagcggcga
     5581 tttggaaacg gcagagaagg tactggaaaa agaacttctg gcctggcagg agaaactgca
     5641 tcagccgatt atcatcaccg aatacggcgt ggatacgtta gccgggctgc actcaatgta
     5701 caccgacatg tggagtgaag agtatcagtg tgcatggctg gatatgtatc accgcgtctt
     5761 tgatcgcgtc agcgccgtcg tcggtgaaca ggtatggaat ttcgccgatt ttgcgacctc
     5821 gcaaggcata ttgcgcgttg gcggtaacaa gaaagggatc ttcactcgcg accgcaaacc
     5881 gaagtcggcg gcttttctgc tgcaaaaacg ctggactggc atgaacttcg gtgaaaaacc
     5941 gcagcaggga ggcaaacaat gaatcaacaa ctctcctggc gcaccggatc ctataagttt
     6001 taataaaact ttaaatagaa aaaggcttct ctcatgagaa gtctttttta tttaaaataa
     6061 atataaaata aaatagaggc tataaatagc ctctatttta tgtgagaaat ccctaaataa
     6121 aaagatgcca gtgtgctgga attcgccctt aggctgcgct agggacctct ttagctcctt
     6181 ggaagctgtc agtagtaggg gatcggtctt gccttgctcg tcggtgatgt acttcaccag
     6241 ctccgcgaag tcgctcttct tgatggagcg catggggacg tgcttggcaa tcacgcgcac
     6301 cccccggccg ttttagcggc taaaaaagtc atggctctgc cctcgggcgg accacgccca
     6361 tcatgacctt gccaagctcg tcctgcttct cttcgatctt cgccagcagg gcgaggatcg
     6421 tggcatcacc gaaccgcgcc gtgcgcgggt cgtcggtgag ccagagtttc agcaggccgc
     6481 ccaggcggcc caggtcgcca ttgatgcggg ccagctcgcg gacgtgctca tagtccacga
     6541 cgcccgtgat tttgtagccc tggccgacgg ccagcaggta ggccgacagg ctcatgccgg
     6601 ccgccgccgc cttttcctca atcgctcttc gttcgtctgg aaggcagtac accttgatag
     6661 gtgggctgcc cttcctggtt ggcttggttt catcagccat ccgcttgccc tcatctgtta
     6721 cgccggcggt agccggccag cctcgcagag caggattccc gttgagcacc gccaggtgcg
     6781 aataagggac agtgaagaag gaacacccgc tcgcgggtgg gcctacttca cctatcctgc
     6841 ccggctgacg ccgttggata caccaaggaa agtctacacg aaccctttgg caaaatcctg
     6901 tatatcgtgc gaaaaaggat ggatataccg aaaaaatcgc tataatgacc ccgaagcagg
     6961 aagggcgaat tctgcagatc tgaccggtct ctgaaaatat aaaaaccaca gattgatact
     7021 aaaaccttgg ttgtgttgct tttcggggct taaatcaagg aaaaatcctt gttttaagcc
     7081 tttcaaaaag aaacacaagg tctttgtact aacctgtggt tatgtataaa attgtagatt
     7141 ttagggtaac aaaaaacacc gtatttctac gatgtttttg cttaaatact tgtttttagt
     7201 tacagacaaa cctgaagtta actatttatc aattcctgca attcgtttac aaaacggcaa
     7261 atgtgaaatc cgtcacatac tgcgtgatga acttgaattg ccaaaggaag tataattttg
     7321 ttatcttctt tataatattt ccccatagta aaaataggaa tcaaataatc atatcctttc
     7381 tgcaaattca gattaaagcc atcgaaggtt gaccacggta tcatagatac attaaaaatg
     7441 ttttccggag catttggctt tccttccatt ctatgattgt ttccataccg ttgcgtatca
     7501 ctttcataat ctgctaaaaa tgatttaaag tcagacttac actcagtcca aaggctggaa
     7561 aatgtttcag tatcattgtg aaatattgta tagcttggta tcatctcatc atatatcccc
     7621 aattcaccat cttgattgat tgccgtccta aactctgaat ggcggtttac aatcattgca
     7681 atataataaa gcattgcagg atatagtttc attccctttt cctttatttg tgtgatatcc
     7741 actttaacgg tcatgctgta ggtacaaggt acacttgcaa agtagtggtc aaaatactct
     7801 tttctgttcc aactattttt atcaattttt tcaaatacca tctaagttcc ctctcaaatt
     7861 caagtttatc gctctaatga acaaagatat tataccacat ttttgtgaat ttttcaactt
     7921 gcccacttcg actgcactcc cgacttaata acttcttgaa cacttgccga aaaagaaaaa
     7981 ctgccgggta cgtacccggg gatcgatccc cggccgagcg ctcttccgct tcctcgctca
     8041 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     8101 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     8161 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     8221 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     8281 tataaagata ccaggcgttt cccccttgaa gctccctcgt gcgctctcct gttccgaccc
     8341 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     8401 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     8461 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     8521 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     8581 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     8641 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     8701 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     8761 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     8821 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaaa tgcaagtttc
     8881 taactaacat acatcatagt tactaaacta tggtaactat aattttatta actatattaa
     8941 gcactgatta gtactataac tcaatataag catatcccct gtatcgtaac tagagaacca
     9001 aacgacggaa aaagcgatat agataagttt agata
//
