LOCUS       sequence-205145-        8188 bp    DNA     circular SYN 14-APR-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 8188)
  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 8188)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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..8188
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3047..4858
                     /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(5103..5474)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            5507..5616
                     /note="incP origin of transfer"
     primer_bind     complement(6807..6824)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6978..7566)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7058..7077)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgcaattctc gtacaattat tttaccaaaa gaaattttta ttaattagtt tttttgcata
       61 aaaataggac tagaaaatca attctaatcc tatcaaaaat ctacttgttt aatctatatg
      121 tatagtacat aaatcctcta tcatttattt tagtttccca tacatctaaa attttagtat
      181 ctaataaaga ttctatcata tttgaaattc tttgctgtgt tactcctaaa tcttttgcta
      241 attcctcttg attaattctt aatatatttc cttttatttt atcatctttg accaataaat
      301 cttgcttata tctcatatga gtatataatc ttaattcttc agttgaaatt attccccaaa
      361 ttactgccat ggtaacaaag taactaatag tatattgttt atctatatta catctaactt
      421 cttttacatt gtatgtatct tttattccta gctttgtaac accttttgtt tttgtaatat
      481 aatcattatg ttgaagttct tttaatgttt ctcttaatgt cttttcactc ataacaatgt
      541 ttttaacttt cttcttacgt ttataggtta tcagctccat tatatcgtct atgtttaatt
      601 ctctatcttt attgttaagt aacacattat agataaacaa ttgattacca gtcattattt
      661 taaccccttt cctattctta tatttcaaat cctttgagtg cttatgtggc atattgaaca
      721 aagtatcttc atcactatat ataaaatctg attctatttt attccaacaa tcactattaa
      781 actcacaacc tgtacaacca aaattaaccg ttttaagctt ctcatacata tacttgactt
      841 gatattctag ctcactattt gaaagactat tttgtgatat atagttccat tctctagcaa
      901 cggatattac ttgacttaaa cttttattcc tcaatcttaa agtcacaact attttttgca
      961 aatctaggtt tttatgtcca tctttactac cattttttaa aatatctact atacaatttg
     1021 gtagcttact atctaatata tattttgtat tagtattacc tactgttttt acatctttgc
     1081 aatatttttt agctaacttt tctatgtcat atctatatat atttttgtct tggtgtatta
     1141 tttttacttg tttagtagta tttttaatat tatatgtata aggtactctt aatacttgag
     1201 tagataaatt tgcttgttta tctgcaccca atttatttat taatgtattt gtaacttctg
     1261 atactaactt aatgttatta gttttattaa ttagcacata aacatggaat ccatttccac
     1321 tatcaactag gatatgataa tgtaatccta tcttagtaaa taaattaatt atatctttat
     1381 aattaaaacc ttgtcctaat tcttttttgt caaaatcaaa tcctaatgca tatctattta
     1441 ttaagttttc attggttcct gcttttccat cagttgtaca taggttaaaa aatgtatttt
     1501 cataataaat attttttgcg aaggtaaatt ctactatatc atctatattt ttaaaaaaag
     1561 ttgttttact atattcttta aattcttctc cgagttctaa ttgtttacta ttacttttat
     1621 tattttgaaa tactctaaca tattcattat caataagatt atttccttgt cctataccat
     1681 ctctaaatac tgccattaga aattttttca acttatattt tgaatctaat tgttccatat
     1741 atttattatc tccttatcta gtttttttaa gagataacac tacattaaat atatacttat
     1801 tgactaatca acttttacaa ctattgaata attttagctt agtatgtata ataatattgt
     1861 taagatgcta aaattgatag caatattatt atataactac agatacttaa tcagttagac
     1921 cgatattgga gttggtagct ccataaactc aattggtagt tgagttgtca ttgtctagta
     1981 gtattatctc attttgttgt tatcttaaaa tattatttta aaaatactct gcttggtcgg
     2041 cttagggtat tttttttgtt taattccaat attggttcaa actcctgtgt aatacttttg
     2101 acacttccat ctgtattgaa ttgacagttg atatggtatc tcattattgg cttgttctta
     2161 tgtttttcta tgtatttatt ttttactttc aaataatttg tatttccatt tttagttatg
     2221 tttaacacac tatccttatt actcaataaa attatttttt gttttaacat atgatttata
     2281 gaatctaaaa attgatattg acttataata gtactttcta cctctctagc tttttctagt
     2341 gtttcaaatt cttcaaactc tgctatattg ctatagtata gcttatcatt aataatatta
     2401 tctgacaact ttaattccat tttataaaac aattccaagt tatgtataaa atttttaatg
     2461 ttacaacata tagacttatt atttgttctt atatggtatc tccatttact acttctagct
     2521 ctcatgttat atgcacaact actcccaaca taaacatata ttgcttgttc taactcatat
     2581 ttttgtaaaa tttcaacatc atatccagtc aaatcgtact ctctcttata tacaaatggt
     2641 ttttctaatt ttttgttcaa taaatctcct cctatttctt acttaattta gctaaaaatc
     2701 tatttatatt tcacctcctt tttgacttta agcctacgaa taccatattt atgattataa
     2761 catcttctat ttttcccaaa tccttacatc tcccctaatt tttagactta agggcgaatt
     2821 ctgcatcaag ctagcttggt accgcggccg cattattaaa aaaaattaca attttgttac
     2881 ttttttgtat acataaatat taaaaaaata aaatataaga aaaagtttat atcttttggt
     2941 taattattac aataagtctc atttattgaa ataatatcaa atatatatta taattggtaa
     3001 ataaggaaaa aataataaga gctcctgcag taaaggagaa aattttatgt tacgtcctgt
     3061 agaaacccca acccgtgaaa tcaaaaaact cgacggcctg tgggcattca gtctggatcg
     3121 cgaaaactgt ggaattgatc agcgttggtg ggaaagcgcg ttacaagaaa gccgggcaat
     3181 tgctgtgcca ggcagtttta acgatcagtt cgccgatgca gatattcgta attatgcggg
     3241 caacgtctgg tatcagcgcg aagtctttat accgaaaggt tgggcaggcc agcgtatcgt
     3301 gctgcgtttc gatgcggtca ctcattacgg caaagtgtgg gtcaataatc aggaagtgat
     3361 ggagcatcag ggcggctata cgccatttga agccgatgtc acgccgtatg ttattgccgg
     3421 gaaaagtgta cgtatcaccg tttgtgtgaa caacgaactg aactggcaga ctatcccgcc
     3481 gggaatggtg attaccgacg aaaacggcaa gaaaaagcag tcttacttcc atgatttctt
     3541 taactatgcc gggatacatc gcagcgtaat gctctacacc acgccgaaca cctgggtgga
     3601 cgatatcacc gtggtgacgc atgtcgcgca agactgtaac cacgcgtctg ttgactggca
     3661 ggtggtggcc aatggtgatg tcagcgttga actgcgtgat gcggatcaac aggtggttgc
     3721 aactggacaa ggcactagcg ggactttgca agtggtgaat ccgcacctct ggcaaccggg
     3781 tgaaggttat ctctatgaac tgtgcgtcac agccaaaagc cagacagagt gtgatatcta
     3841 cccgcttcgc gtcggcatcc ggtcagtggc agtgaagggc gaacagttcc tgattaacca
     3901 caaaccgttc tactttactg gctttggtcg tcatgaagat gcggacttgc gtggcaaagg
     3961 attcgataac gtgctgatgg tgcacgacca cgcattaatg gactggattg gggccaactc
     4021 ctaccgtacc tcgcattacc cttacgctga agagatgctc gactgggcag atgaacatgg
     4081 catcgtggtg attgatgaaa ctgctgctgt cggctttaac ctctctttag gcattggttt
     4141 cgaagcgggc aacaagccga aagaactgta cagcgaagag gcagtcaacg gggaaactca
     4201 gcaagcgcac ttacaggcga ttaaagagct gatagcgcgt gacaaaaacc acccaagcgt
     4261 ggtgatgtgg agtattgcca acgaaccgga tacccgtccg caaggtgcac gggaatattt
     4321 cgcgccactg gcggaagcaa cgcgtaaact cgacccgacg cgtccgatca cctgcgtcaa
     4381 tgtaatgttc tgcgacgctc acaccgatac catcagcgat ctctttgatg tgctgtgcct
     4441 gaaccgttat tacggatggt atgtccaaag cggcgatttg gaaacggcag agaaggtact
     4501 ggaaaaagaa cttctggcct ggcaggagaa actgcatcag ccgattatca tcaccgaata
     4561 cggcgtggat acgttagccg ggctgcactc aatgtacacc gacatgtgga gtgaagagta
     4621 tcagtgtgca tggctggata tgtatcaccg cgtctttgat cgcgtcagcg ccgtcgtcgg
     4681 tgaacaggta tggaatttcg ccgattttgc gacctcgcaa ggcatattgc gcgttggcgg
     4741 taacaagaaa gggatcttca ctcgcgaccg caaaccgaag tcggcggctt ttctgctgca
     4801 aaaacgctgg actggcatga acttcggtga aaaaccgcag cagggaggca aacaatgaat
     4861 caacaactct cctggcgcac cggatccact agtaacggcc gccagtgtgc tggaattcgc
     4921 ccttaggctg cgctagggac ctctttagct ccttggaagc tgtcagtagt aggggatcgg
     4981 tcttgccttg ctcgtcggtg atgtacttca ccagctccgc gaagtcgctc ttcttgatgg
     5041 agcgcatggg gacgtgcttg gcaatcacgc gcaccccccg gccgttttag cggctaaaaa
     5101 agtcatggct ctgccctcgg gcggaccacg cccatcatga ccttgccaag ctcgtcctgc
     5161 ttctcttcga tcttcgccag cagggcgagg atcgtggcat caccgaaccg cgccgtgcgc
     5221 gggtcgtcgg tgagccagag tttcagcagg ccgcccaggc ggcccaggtc gccattgatg
     5281 cgggccagct cgcggacgtg ctcatagtcc acgacgcccg tgattttgta gccctggccg
     5341 acggccagca ggtaggccga caggctcatg ccggccgccg ccgccttttc ctcaatcgct
     5401 cttcgttcgt ctggaaggca gtacaccttg ataggtgggc tgcccttcct ggttggcttg
     5461 gtttcatcag ccatccgctt gccctcatct gttacgccgg cggtagccgg ccagcctcgc
     5521 agagcaggat tcccgttgag caccgccagg tgcgaataag ggacagtgaa gaaggaacac
     5581 ccgctcgcgg gtgggcctac ttcacctatc ctgcccggct gacgccgttg gatacaccaa
     5641 ggaaagtcta cacgaaccct ttggcaaaat cctgtatatc gtgcgaaaaa ggatggatat
     5701 accgaaaaaa tcgctataat gaccccgaag caggaagggc gaattctgca gatctgaccg
     5761 gtctctgaaa atataaaaac cacagattga tactaaaacc ttggttgtgt tgcttttcgg
     5821 ggcttaaatc aaggaaaaat ccttgtttta agcctttcaa aaagaaacac aaggtctttg
     5881 tactaacctg tggttatgta taaaattgta gattttaggg taacaaaaaa caccgtattt
     5941 ctacgatgtt tttgcttaaa tacttgtttt tagttacaga caaacctgaa gttaactatt
     6001 tatcaattcc tgcaattcgt ttacaaaacg gcaaatgtga aatccgtcac atactgcgtg
     6061 atgaacttga attgccaaag gaagtataat tttgttatct tctttataat atttccccat
     6121 agtaaaaata ggaatcaaat aatcatatcc tttctgcaaa ttcagattaa agccatcgaa
     6181 ggttgaccac ggtatcatag atacattaaa aatgttttcc ggagcatttg gctttccttc
     6241 cattctatga ttgtttccat accgttgcgt atcactttca taatctgcta aaaatgattt
     6301 aaagtcagac ttacactcag tccaaaggct ggaaaatgtt tcagtatcat tgtgaaatat
     6361 tgtatagctt ggtatcatct catcatatat ccccaattca ccatcttgat tgattgccgt
     6421 cctaaactct gaatggcggt ttacaatcat tgcaatataa taaagcattg caggatatag
     6481 tttcattccc ttttccttta tttgtgtgat atccacttta acggtcatgc tgtaggtaca
     6541 aggtacactt gcaaagtagt ggtcaaaata ctcttttctg ttccaactat ttttatcaat
     6601 tttttcaaat accatctaag ttccctctca aattcaagtt tatcgctcta atgaacaaag
     6661 atattatacc acatttttgt gaatttttca acttgcccac ttcgactgca ctcccgactt
     6721 aataacttct tgaacacttg ccgaaaaaga aaaactgccg ggtacgtacc cggggatcga
     6781 tccccggccg agcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6841 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6901 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6961 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     7021 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     7081 tgaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     7141 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     7201 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     7261 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     7321 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     7381 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     7441 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     7501 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     7561 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7621 cgttaaggga ttttggtcat gaaatgcaag tttctaacta acatacatca tagttactaa
     7681 actatggtaa ctataatttt attaactata ttaagcactg attagtacta taactcaata
     7741 taagcatatc ccctgtatcg taactagaga accaaacgac ggaaaaagcg atatagataa
     7801 gtttagatag tgcaccaatt ttttactttt ctattttaga atctagcatt tccaatgctt
     7861 actaatacta tttttgattt ttaacttgtt ttaaaaaagc acctactaat taggattcta
     7921 gctctatatt ctagttatac attctctata ttctagctct atattctagt tatacattct
     7981 ctatattcta gctctatatt ctagttatac attctctata ttctagctct atattctagt
     8041 tatacattct ctatattcta gctctatatt ctagttatac attctctaaa aaacatatat
     8101 atatatacta catatatatg tacaattatt ttaccaaaag aaatttttaa aacggcttaa
     8161 tttcaatact ttgatgccat tttttttt
//
