LOCUS       sequence-229725-        8513 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli-C. difficile shuttle vector for xylose-inducible expression 
            in C. difficile; Pxyl driving expression of red fluorescent protein 
            (mCherryOpt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8513)
  AUTHORS   Muh U, Pannullo AG, Weiss DS, Ellermeier CD
  TITLE     A xylose-inducible expression system and a CRISPRi-plasmid for 
            targeted knock-down of gene expression in Clostridioides difficile.
  JOURNAL   J Bacteriol. 2019 Feb 11. pii: JB.00711-18. doi: 
            10.1128/JB.00711-18.
  PUBMED    30745377
REFERENCE   2  (bases 1 to 8513)
  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: "J 
            Bacteriol. 2019 Feb 11. pii: JB.00711-18. doi: 10.1128/JB.00711-18."
COMMENT     Sequence Label: pAP114
FEATURES             Location/Qualifiers
     source          1..8513
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(2631..3002)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            3035..3144
                     /note="incP origin of transfer"
     primer_bind     complement(4335..4352)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4506..5094)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4586..4605)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttaagcctac gaataccata tttatgatta taacatcttc tatttttccc aaatccttac
       61 atctccccta atttttagac ttaagggcga attctgcatc aagctagcat aaaaataaga
      121 agcctgcatt tgcaggcttc ttatttttat ggtacgtata atttaaaggg ttatggctgt
      181 atatagtcat aacccttata tttcattaat taaagttaaa ttttgtatat tcaaaaagct
      241 cttgatattt acggcagagc caccatatag agatgcctca gaccctaatg atgaaatttc
      301 tatatttatt cctttgttaa aagagctaac taacatatct tttgtaatct gtaatatctc
      361 agggatatca ctaattattt ggctatttaa atatattatt tcaggagcat aagttgtaat
      421 agcattattt atagctattg ttaagtaact acaaaactca tgaataactt ttttggcatt
      481 ctggttatct tcataatata gttgctttac tatatcagaa tctattttag gaatattttc
      541 taaagaagaa agctgttcaa ataccttttt ttctgaacaa tactgttcta aacagccacg
      601 atttccacaa ggacatagtt taccattagg catgatgata gtgtgaccaa tttctccact
      661 cataccattt ctgccactat ataatttatt atttattata atgcctgaac caaatccact
      721 gtgaatactg agactaagta gtgagttatg tacagttgaa aaagtattct cagctaaagc
      781 tgttaagttt gcttcatttt ctatgtgaat aggaaagtca tactttttgc taaggatgct
      841 atataagtca atctcattaa gattgtaata tggagtaaac aatactttat tttcacaagt
      901 aatcccatgt atagccaaag taagaccaat taccttataa ggagtgtcta ttttggatat
      961 gttataacta tttataattt catctatcaa ttgtataaca ttgtctttac ttacttgtat
     1021 atctgttaat ttcttagaat ttataatagt tccatctaag taagatagag aagaaaatat
     1081 atagtcgtat ccaatgtcca tacttaaaga aatccctgca catttattaa atactaataa
     1141 tatgggtttt cttccgccac tatgagtact atttccaatt cctatttcat gaactagaga
     1201 ttcatcaata agttttttgg taatagcgga tatagttgct ttattcaatc ctatagtaga
     1261 agctatactt gccctagaaa taggaccatt ttttataatt tgttcaagta ccaatctttc
     1321 attcatttct cgaatagtgt atttatcagt aaccaatttg atattcctcc ttaaaataat
     1381 attgtaatac tttttacaca aaaataaaag gttattttgc attgacaaag ataattaaat
     1441 attttattat tagttcataa gttagtttaa tatactaaca aaaataaagc aagtaaaata
     1501 tacctaaaat ataaaaaaat taggatagga aaacgatagt tatgaagtga gctcctgcag
     1561 taaaggagaa aattttatgg tatctaaagg agaagaagat aatatggcta taattaaaga
     1621 atttatgaga tttaaagttc atatggaagg atctgttaat ggacatgaat ttgaaataga
     1681 aggagaagga gaaggaagac cttatgaagg tactcaaaca gctaaattaa aagtaactaa
     1741 aggtggtcca ttaccatttg catgggatat attaagtcct caatttatgt atggatctaa
     1801 agcatatgtt aaacatcctg ctgatatacc tgattattta aaattatcat ttccagaagg
     1861 ttttaaatgg gaaagagtta tgaattttga agatggtgga gtagttactg taacacaaga
     1921 tagttcatta caagatggag attttatata taaagttaaa ttaagaggta ctaattttcc
     1981 atctgatgga ccagtaatgc aaaagaaaac tatgggatgg gaagcatcat ctgaaagaat
     2041 gtatccagaa gatggagcat taaaaggtga aattaaacaa agattaaaat taaaagatgg
     2101 aggtcattat gatgctgaag ttaagactac atataaagct aagaaaccag ttcaattacc
     2161 aggtgcatat aatgttaata ttaaattaga tataacttct cataatgaag attatactat
     2221 agttgaacaa tatgaaagag cagaaggaag acattcaaca ggaggtatgg atgaattata
     2281 taaataagga tcctataagt tttaataaaa ctttaaatag aaaaaggctt ctctcatgag
     2341 aagtcttttt tatttaaaat aaatataaaa taaaatagag gctataaata gcctctattt
     2401 tatgtgagaa atccctaaat aaaaagatgc cagtgtgctg gaattcgccc ttaggctgcg
     2461 ctagggacct ctttagctcc ttggaagctg tcagtagtag gggatcggtc ttgccttgct
     2521 cgtcggtgat gtacttcacc agctccgcga agtcgctctt cttgatggag cgcatgggga
     2581 cgtgcttggc aatcacgcgc accccccggc cgttttagcg gctaaaaaag tcatggctct
     2641 gccctcgggc ggaccacgcc catcatgacc ttgccaagct cgtcctgctt ctcttcgatc
     2701 ttcgccagca gggcgaggat cgtggcatca ccgaaccgcg ccgtgcgcgg gtcgtcggtg
     2761 agccagagtt tcagcaggcc gcccaggcgg cccaggtcgc cattgatgcg ggccagctcg
     2821 cggacgtgct catagtccac gacgcccgtg attttgtagc cctggccgac ggccagcagg
     2881 taggccgaca ggctcatgcc ggccgccgcc gccttttcct caatcgctct tcgttcgtct
     2941 ggaaggcagt acaccttgat aggtgggctg cccttcctgg ttggcttggt ttcatcagcc
     3001 atccgcttgc cctcatctgt tacgccggcg gtagccggcc agcctcgcag agcaggattc
     3061 ccgttgagca ccgccaggtg cgaataaggg acagtgaaga aggaacaccc gctcgcgggt
     3121 gggcctactt cacctatcct gcccggctga cgccgttgga tacaccaagg aaagtctaca
     3181 cgaacccttt ggcaaaatcc tgtatatcgt gcgaaaaagg atggatatac cgaaaaaatc
     3241 gctataatga ccccgaagca ggaagggcga attctgcaga tctgaccggt ctctgaaaat
     3301 ataaaaacca cagattgata ctaaaacctt ggttgtgttg cttttcgggg cttaaatcaa
     3361 ggaaaaatcc ttgttttaag cctttcaaaa agaaacacaa ggtctttgta ctaacctgtg
     3421 gttatgtata aaattgtaga ttttagggta acaaaaaaca ccgtatttct acgatgtttt
     3481 tgcttaaata cttgttttta gttacagaca aacctgaagt taactattta tcaattcctg
     3541 caattcgttt acaaaacggc aaatgtgaaa tccgtcacat actgcgtgat gaacttgaat
     3601 tgccaaagga agtataattt tgttatcttc tttataatat ttccccatag taaaaatagg
     3661 aatcaaataa tcatatcctt tctgcaaatt cagattaaag ccatcgaagg ttgaccacgg
     3721 tatcatagat acattaaaaa tgttttccgg agcatttggc tttccttcca ttctatgatt
     3781 gtttccatac cgttgcgtat cactttcata atctgctaaa aatgatttaa agtcagactt
     3841 acactcagtc caaaggctgg aaaatgtttc agtatcattg tgaaatattg tatagcttgg
     3901 tatcatctca tcatatatcc ccaattcacc atcttgattg attgccgtcc taaactctga
     3961 atggcggttt acaatcattg caatataata aagcattgca ggatatagtt tcattccctt
     4021 ttcctttatt tgtgtgatat ccactttaac ggtcatgctg taggtacaag gtacacttgc
     4081 aaagtagtgg tcaaaatact cttttctgtt ccaactattt ttatcaattt tttcaaatac
     4141 catctaagtt ccctctcaaa ttcaagttta tcgctctaat gaacaaagat attataccac
     4201 atttttgtga atttttcaac ttgcccactt cgactgcact cccgacttaa taacttcttg
     4261 aacacttgcc gaaaaagaaa aactgccggg tacgtacccg gggatcgatc cccggccgag
     4321 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4381 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4441 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4501 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4561 aggtggcgaa acccgacagg actataaaga taccaggcgt ttcccccttg aagctccctc
     4621 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4681 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4741 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4801 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4861 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4921 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     4981 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5041 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5101 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5161 ttggtcatga aatgcaagtt tctaactaac atacatcata gttactaaac tatggtaact
     5221 ataattttat taactatatt aagcactgat tagtactata actcaatata agcatatccc
     5281 ctgtatcgta actagagaac caaacgacgg aaaaagcgat atagataagt ttagatagtg
     5341 caccaatttt ttacttttct attttagaat ctagcatttc caatgcttac taatactatt
     5401 tttgattttt aacttgtttt aaaaaagcac ctactaatta ggattctagc tctatattct
     5461 agttatacat tctctatatt ctagctctat attctagtta tacattctct atattctagc
     5521 tctatattct agttatacat tctctatatt ctagctctat attctagtta tacattctct
     5581 atattctagc tctatattct agttatacat tctctatatt ctagctctat attctagtta
     5641 tacattctct atattctagc tctatattct agttatacat tctctaaaaa acatatatat
     5701 atatactaca tatatatgta caattatttt accaaaagaa atttttaaaa cggcttaatt
     5761 tcaatacttt gatgccattt tttttttgca attctcgtac aattatttta ccaaaagaaa
     5821 tttttattaa ttagtttttt tgcataaaaa taggactaga aaatcaattc taatcctatc
     5881 aaaaatctac ttgtttaatc tatatgtata gtacataaat cctctatcat ttattttagt
     5941 ttcccataca tctaaaattt tagtatctaa taaagattct atcatatttg aaattctttg
     6001 ctgtgttact cctaaatctt ttgctaattc ctcttgatta attcttaata tatttccttt
     6061 tattttatca tctttgacca ataaatcttg cttatatctc atatgagtat ataatcttaa
     6121 ttcttcagtt gaaattattc cccaaattac tgccatggta acaaagtaac taatagtata
     6181 ttgtttatct atattacatc taacttcttt tacattgtat gtatctttta ttcctagctt
     6241 tgtaacacct tttgtttttg taatataatc attatgttga agttctttta atgtttctct
     6301 taatgtcttt tcactcataa caatgttttt aactttcttc ttacgtttat aggttatcag
     6361 ctccattata tcgtctatgt ttaattctct atctttattg ttaagtaaca cattatagat
     6421 aaacaattga ttaccagtca ttattttaac ccctttccta ttcttatatt tcaaatcctt
     6481 tgagtgctta tgtggcatat tgaacaaagt atcttcatca ctatatataa aatctgattc
     6541 tattttattc caacaatcac tattaaactc acaacctgta caaccaaaat taaccgtttt
     6601 aagcttctca tacatatact tgacttgata ttctagctca ctatttgaaa gactattttg
     6661 tgatatatag ttccattctc tagcaacgga tattacttga cttaaacttt tattcctcaa
     6721 tcttaaagtc acaactattt tttgcaaatc taggttttta tgtccatctt tactaccatt
     6781 ttttaaaata tctactatac aatttggtag cttactatct aatatatatt ttgtattagt
     6841 attacctact gtttttacat ctttgcaata ttttttagct aacttttcta tgtcatatct
     6901 atatatattt ttgtcttggt gtattatttt tacttgttta gtagtatttt taatattata
     6961 tgtataaggt actcttaata cttgagtaga taaatttgct tgtttatctg cacccaattt
     7021 atttattaat gtatttgtaa cttctgatac taacttaatg ttattagttt tattaattag
     7081 cacataaaca tggaatccat ttccactatc aactaggata tgataatgta atcctatctt
     7141 agtaaataaa ttaattatat ctttataatt aaaaccttgt cctaattctt ttttgtcaaa
     7201 atcaaatcct aatgcatatc tatttattaa gttttcattg gttcctgctt ttccatcagt
     7261 tgtacatagg ttaaaaaatg tattttcata ataaatattt tttgcgaagg taaattctac
     7321 tatatcatct atatttttaa aaaaagttgt tttactatat tctttaaatt cttctccgag
     7381 ttctaattgt ttactattac ttttattatt ttgaaatact ctaacatatt cattatcaat
     7441 aagattattt ccttgtccta taccatctct aaatactgcc attagaaatt ttttcaactt
     7501 atattttgaa tctaattgtt ccatatattt attatctcct tatctagttt ttttaagaga
     7561 taacactaca ttaaatatat acttattgac taatcaactt ttacaactat tgaataattt
     7621 tagcttagta tgtataataa tattgttaag atgctaaaat tgatagcaat attattatat
     7681 aactacagat acttaatcag ttagaccgat attggagttg gtagctccat aaactcaatt
     7741 ggtagttgag ttgtcattgt ctagtagtat tatctcattt tgttgttatc ttaaaatatt
     7801 attttaaaaa tactctgctt ggtcggctta gggtattttt tttgtttaat tccaatattg
     7861 gttcaaactc ctgtgtaata cttttgacac ttccatctgt attgaattga cagttgatat
     7921 ggtatctcat tattggcttg ttcttatgtt tttctatgta tttatttttt actttcaaat
     7981 aatttgtatt tccattttta gttatgttta acacactatc cttattactc aataaaatta
     8041 ttttttgttt taacatatga tttatagaat ctaaaaattg atattgactt ataatagtac
     8101 tttctacctc tctagctttt tctagtgttt caaattcttc aaactctgct atattgctat
     8161 agtatagctt atcattaata atattatctg acaactttaa ttccatttta taaaacaatt
     8221 ccaagttatg tataaaattt ttaatgttac aacatataga cttattattt gttcttatat
     8281 ggtatctcca tttactactt ctagctctca tgttatatgc acaactactc ccaacataaa
     8341 catatattgc ttgttctaac tcatattttt gtaaaatttc aacatcatat ccagtcaaat
     8401 cgtactctct cttatataca aatggttttt ctaatttttt gttcaataaa tctcctccta
     8461 tttcttactt aatttagcta aaaatctatt tatatttcac ctcctttttg act
//
