LOCUS       sequence-201424-        8258 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C. difficile constitutive expression system. Shuttle plasmid 
            containing constitutive Pcwp2 -gusA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8258)
  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 8258)
  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: pRPF144
FEATURES             Location/Qualifiers
     source          1..8258
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3496..5307
                     /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(5552..5923)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            5956..6065
                     /note="incP origin of transfer"
     primer_bind     complement(7256..7273)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7427..8015)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7507..7526)
                     /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 tagcttggta ccgcggccgc
     3301 attattaaaa aaaattacaa ttttgttact tttttgtata cataaatatt aaaaaaataa
     3361 aatataagaa aaagtttata tcttttggtt aattattaca ataagtctca tttattgaaa
     3421 taatatcaaa tatatattat aattggtaaa taaggaaaaa ataataagag ctcctgcagt
     3481 aaaggagaaa attttatgtt acgtcctgta gaaaccccaa cccgtgaaat caaaaaactc
     3541 gacggcctgt gggcattcag tctggatcgc gaaaactgtg gaattgatca gcgttggtgg
     3601 gaaagcgcgt tacaagaaag ccgggcaatt gctgtgccag gcagttttaa cgatcagttc
     3661 gccgatgcag atattcgtaa ttatgcgggc aacgtctggt atcagcgcga agtctttata
     3721 ccgaaaggtt gggcaggcca gcgtatcgtg ctgcgtttcg atgcggtcac tcattacggc
     3781 aaagtgtggg tcaataatca ggaagtgatg gagcatcagg gcggctatac gccatttgaa
     3841 gccgatgtca cgccgtatgt tattgccggg aaaagtgtac gtatcaccgt ttgtgtgaac
     3901 aacgaactga actggcagac tatcccgccg ggaatggtga ttaccgacga aaacggcaag
     3961 aaaaagcagt cttacttcca tgatttcttt aactatgccg ggatacatcg cagcgtaatg
     4021 ctctacacca cgccgaacac ctgggtggac gatatcaccg tggtgacgca tgtcgcgcaa
     4081 gactgtaacc acgcgtctgt tgactggcag gtggtggcca atggtgatgt cagcgttgaa
     4141 ctgcgtgatg cggatcaaca ggtggttgca actggacaag gcactagcgg gactttgcaa
     4201 gtggtgaatc cgcacctctg gcaaccgggt gaaggttatc tctatgaact gtgcgtcaca
     4261 gccaaaagcc agacagagtg tgatatctac ccgcttcgcg tcggcatccg gtcagtggca
     4321 gtgaagggcg aacagttcct gattaaccac aaaccgttct actttactgg ctttggtcgt
     4381 catgaagatg cggacttgcg tggcaaagga ttcgataacg tgctgatggt gcacgaccac
     4441 gcattaatgg actggattgg ggccaactcc taccgtacct cgcattaccc ttacgctgaa
     4501 gagatgctcg actgggcaga tgaacatggc atcgtggtga ttgatgaaac tgctgctgtc
     4561 ggctttaacc tctctttagg cattggtttc gaagcgggca acaagccgaa agaactgtac
     4621 agcgaagagg cagtcaacgg ggaaactcag caagcgcact tacaggcgat taaagagctg
     4681 atagcgcgtg acaaaaacca cccaagcgtg gtgatgtgga gtattgccaa cgaaccggat
     4741 acccgtccgc aaggtgcacg ggaatatttc gcgccactgg cggaagcaac gcgtaaactc
     4801 gacccgacgc gtccgatcac ctgcgtcaat gtaatgttct gcgacgctca caccgatacc
     4861 atcagcgatc tctttgatgt gctgtgcctg aaccgttatt acggatggta tgtccaaagc
     4921 ggcgatttgg aaacggcaga gaaggtactg gaaaaagaac ttctggcctg gcaggagaaa
     4981 ctgcatcagc cgattatcat caccgaatac ggcgtggata cgttagccgg gctgcactca
     5041 atgtacaccg acatgtggag tgaagagtat cagtgtgcat ggctggatat gtatcaccgc
     5101 gtctttgatc gcgtcagcgc cgtcgtcggt gaacaggtat ggaatttcgc cgattttgcg
     5161 acctcgcaag gcatattgcg cgttggcggt aacaagaaag ggatcttcac tcgcgaccgc
     5221 aaaccgaagt cggcggcttt tctgctgcaa aaacgctgga ctggcatgaa cttcggtgaa
     5281 aaaccgcagc agggaggcaa acaatgaatc aacaactctc ctggcgcacc ggatccacta
     5341 gtaacggccg ccagtgtgct ggaattcgcc cttaggctgc gctagggacc tctttagctc
     5401 cttggaagct gtcagtagta ggggatcggt cttgccttgc tcgtcggtga tgtacttcac
     5461 cagctccgcg aagtcgctct tcttgatgga gcgcatgggg acgtgcttgg caatcacgcg
     5521 caccccccgg ccgttttagc ggctaaaaaa gtcatggctc tgccctcggg cggaccacgc
     5581 ccatcatgac cttgccaagc tcgtcctgct tctcttcgat cttcgccagc agggcgagga
     5641 tcgtggcatc accgaaccgc gccgtgcgcg ggtcgtcggt gagccagagt ttcagcaggc
     5701 cgcccaggcg gcccaggtcg ccattgatgc gggccagctc gcggacgtgc tcatagtcca
     5761 cgacgcccgt gattttgtag ccctggccga cggccagcag gtaggccgac aggctcatgc
     5821 cggccgccgc cgccttttcc tcaatcgctc ttcgttcgtc tggaaggcag tacaccttga
     5881 taggtgggct gcccttcctg gttggcttgg tttcatcagc catccgcttg ccctcatctg
     5941 ttacgccggc ggtagccggc cagcctcgca gagcaggatt cccgttgagc accgccaggt
     6001 gcgaataagg gacagtgaag aaggaacacc cgctcgcggg tgggcctact tcacctatcc
     6061 tgcccggctg acgccgttgg atacaccaag gaaagtctac acgaaccctt tggcaaaatc
     6121 ctgtatatcg tgcgaaaaag gatggatata ccgaaaaaat cgctataatg accccgaagc
     6181 aggaagggcg aattctgcag atctgaccgg tctctgaaaa tataaaaacc acagattgat
     6241 actaaaacct tggttgtgtt gcttttcggg gcttaaatca aggaaaaatc cttgttttaa
     6301 gcctttcaaa aagaaacaca aggtctttgt actaacctgt ggttatgtat aaaattgtag
     6361 attttagggt aacaaaaaac accgtatttc tacgatgttt ttgcttaaat acttgttttt
     6421 agttacagac aaacctgaag ttaactattt atcaattcct gcaattcgtt tacaaaacgg
     6481 caaatgtgaa atccgtcaca tactgcgtga tgaacttgaa ttgccaaagg aagtataatt
     6541 ttgttatctt ctttataata tttccccata gtaaaaatag gaatcaaata atcatatcct
     6601 ttctgcaaat tcagattaaa gccatcgaag gttgaccacg gtatcataga tacattaaaa
     6661 atgttttccg gagcatttgg ctttccttcc attctatgat tgtttccata ccgttgcgta
     6721 tcactttcat aatctgctaa aaatgattta aagtcagact tacactcagt ccaaaggctg
     6781 gaaaatgttt cagtatcatt gtgaaatatt gtatagcttg gtatcatctc atcatatatc
     6841 cccaattcac catcttgatt gattgccgtc ctaaactctg aatggcggtt tacaatcatt
     6901 gcaatataat aaagcattgc aggatatagt ttcattccct tttcctttat ttgtgtgata
     6961 tccactttaa cggtcatgct gtaggtacaa ggtacacttg caaagtagtg gtcaaaatac
     7021 tcttttctgt tccaactatt tttatcaatt ttttcaaata ccatctaagt tccctctcaa
     7081 attcaagttt atcgctctaa tgaacaaaga tattatacca catttttgtg aatttttcaa
     7141 cttgcccact tcgactgcac tcccgactta ataacttctt gaacacttgc cgaaaaagaa
     7201 aaactgccgg gtacgtaccc ggggatcgat ccccggccga gcgctcttcc gcttcctcgc
     7261 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     7321 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     7381 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     7441 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     7501 gactataaag ataccaggcg tttccccctt gaagctccct cgtgcgctct cctgttccga
     7561 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     7621 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     7681 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     7741 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     7801 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     7861 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     7921 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     7981 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     8041 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg aaatgcaagt
     8101 ttctaactaa catacatcat agttactaaa ctatggtaac tataatttta ttaactatat
     8161 taagcactga ttagtactat aactcaatat aagcatatcc cctgtatcgt aactagagaa
     8221 ccaaacgacg gaaaaagcga tatagataag tttagata
//
