LOCUS       Exported                8513 bp ds-DNA     circular SYN 12-MAY-2021
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    pAP114
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 May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8513
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      623..1211
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1112..1131
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1365..1382
                     /label=L4440
                     /note="L4440 vector, forward primer"
     oriT            2573..2682
                     /note="incP origin of transfer"
     CDS             2715..3086
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
ORIGIN
        1 tatatatgta gtatatatat atatgttttt tagagaatgt ataactagaa tatagagcta
       61 gaatatagag aatgtataac tagaatatag agctagaata tagagaatgt ataactagaa
      121 tatagagcta gaatatagag aatgtataac tagaatatag agctagaata tagagaatgt
      181 ataactagaa tatagagcta gaatatagag aatgtataac tagaatatag agctagaata
      241 tagagaatgt ataactagaa tatagagcta gaatcctaat tagtaggtgc ttttttaaaa
      301 caagttaaaa atcaaaaata gtattagtaa gcattggaaa tgctagattc taaaatagaa
      361 aagtaaaaaa ttggtgcact atctaaactt atctatatcg ctttttccgt cgtttggttc
      421 tctagttacg atacagggga tatgcttata ttgagttata gtactaatca gtgcttaata
      481 tagttaataa aattatagtt accatagttt agtaactatg atgtatgtta gttagaaact
      541 tgcatttcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
      601 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
      661 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
      721 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
      781 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
      841 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
      901 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
      961 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     1021 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     1081 gaacaggaga gcgcacgagg gagcttcaag ggggaaacgc ctggtatctt tatagtcctg
     1141 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     1201 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     1261 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     1321 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     1381 aggaagcgga agagcgctcg gccggggatc gatccccggg tacgtacccg gcagtttttc
     1441 tttttcggca agtgttcaag aagttattaa gtcgggagtg cagtcgaagt gggcaagttg
     1501 aaaaattcac aaaaatgtgg tataatatct ttgttcatta gagcgataaa cttgaatttg
     1561 agagggaact tagatggtat ttgaaaaaat tgataaaaat agttggaaca gaaaagagta
     1621 ttttgaccac tactttgcaa gtgtaccttg tacctacagc atgaccgtta aagtggatat
     1681 cacacaaata aaggaaaagg gaatgaaact atatcctgca atgctttatt atattgcaat
     1741 gattgtaaac cgccattcag agtttaggac ggcaatcaat caagatggtg aattggggat
     1801 atatgatgag atgataccaa gctatacaat atttcacaat gatactgaaa cattttccag
     1861 cctttggact gagtgtaagt ctgactttaa atcattttta gcagattatg aaagtgatac
     1921 gcaacggtat ggaaacaatc atagaatgga aggaaagcca aatgctccgg aaaacatttt
     1981 taatgtatct atgataccgt ggtcaacctt cgatggcttt aatctgaatt tgcagaaagg
     2041 atatgattat ttgattccta tttttactat ggggaaatat tataaagaag ataacaaaat
     2101 tatacttcct ttggcaattc aagttcatca cgcagtatgt gacggatttc acatttgccg
     2161 ttttgtaaac gaattgcagg aattgataaa tagttaactt caggtttgtc tgtaactaaa
     2221 aacaagtatt taagcaaaaa catcgtagaa atacggtgtt ttttgttacc ctaaaatcta
     2281 caattttata cataaccaca ggttagtaca aagaccttgt gtttcttttt gaaaggctta
     2341 aaacaaggat ttttccttga tttaagcccc gaaaagcaac acaaccaagg ttttagtatc
     2401 aatctgtggt ttttatattt tcagagaccg gtcagatctg cagaattcgc ccttcctgct
     2461 tcggggtcat tatagcgatt ttttcggtat atccatcctt tttcgcacga tatacaggat
     2521 tttgccaaag ggttcgtgta gactttcctt ggtgtatcca acggcgtcag ccgggcagga
     2581 taggtgaagt aggcccaccc gcgagcgggt gttccttctt cactgtccct tattcgcacc
     2641 tggcggtgct caacgggaat cctgctctgc gaggctggcc ggctaccgcc ggcgtaacag
     2701 atgagggcaa gcggatggct gatgaaacca agccaaccag gaagggcagc ccacctatca
     2761 aggtgtactg ccttccagac gaacgaagag cgattgagga aaaggcggcg gcggccggca
     2821 tgagcctgtc ggcctacctg ctggccgtcg gccagggcta caaaatcacg ggcgtcgtgg
     2881 actatgagca cgtccgcgag ctggcccgca tcaatggcga cctgggccgc ctgggcggcc
     2941 tgctgaaact ctggctcacc gacgacccgc gcacggcgcg gttcggtgat gccacgatcc
     3001 tcgccctgct ggcgaagatc gaagagaagc aggacgagct tggcaaggtc atgatgggcg
     3061 tggtccgccc gagggcagag ccatgacttt tttagccgct aaaacggccg gggggtgcgc
     3121 gtgattgcca agcacgtccc catgcgctcc atcaagaaga gcgacttcgc ggagctggtg
     3181 aagtacatca ccgacgagca aggcaagacc gatcccctac tactgacagc ttccaaggag
     3241 ctaaagaggt ccctagcgca gcctaagggc gaattccagc acactggcat ctttttattt
     3301 agggatttct cacataaaat agaggctatt tatagcctct attttatttt atatttattt
     3361 taaataaaaa agacttctca tgagagaagc ctttttctat ttaaagtttt attaaaactt
     3421 ataggatcct tatttatata attcatccat acctcctgtt gaatgtcttc cttctgctct
     3481 ttcatattgt tcaactatag tataatcttc attatgagaa gttatatcta atttaatatt
     3541 aacattatat gcacctggta attgaactgg tttcttagct ttatatgtag tcttaacttc
     3601 agcatcataa tgacctccat cttttaattt taatctttgt ttaatttcac cttttaatgc
     3661 tccatcttct ggatacattc tttcagatga tgcttcccat cccatagttt tcttttgcat
     3721 tactggtcca tcagatggaa aattagtacc tcttaattta actttatata taaaatctcc
     3781 atcttgtaat gaactatctt gtgttacagt aactactcca ccatcttcaa aattcataac
     3841 tctttcccat ttaaaacctt ctggaaatga taattttaaa taatcaggta tatcagcagg
     3901 atgtttaaca tatgctttag atccatacat aaattgagga cttaatatat cccatgcaaa
     3961 tggtaatgga ccacctttag ttacttttaa tttagctgtt tgagtacctt cataaggtct
     4021 tccttctcct tctccttcta tttcaaattc atgtccatta acagatcctt ccatatgaac
     4081 tttaaatctc ataaattctt taattatagc catattatct tcttctcctt tagataccat
     4141 aaaattttct cctttactgc aggagctcac ttcataacta tcgttttcct atcctaattt
     4201 ttttatattt taggtatatt ttacttgctt tatttttgtt agtatattaa actaacttat
     4261 gaactaataa taaaatattt aattatcttt gtcaatgcaa aataaccttt tatttttgtg
     4321 taaaaagtat tacaatatta ttttaaggag gaatatcaaa ttggttactg ataaatacac
     4381 tattcgagaa atgaatgaaa gattggtact tgaacaaatt ataaaaaatg gtcctatttc
     4441 tagggcaagt atagcttcta ctataggatt gaataaagca actatatccg ctattaccaa
     4501 aaaacttatt gatgaatctc tagttcatga aataggaatt ggaaatagta ctcatagtgg
     4561 cggaagaaaa cccatattat tagtatttaa taaatgtgca gggatttctt taagtatgga
     4621 cattggatac gactatatat tttcttctct atcttactta gatggaacta ttataaattc
     4681 taagaaatta acagatatac aagtaagtaa agacaatgtt atacaattga tagatgaaat
     4741 tataaatagt tataacatat ccaaaataga cactccttat aaggtaattg gtcttacttt
     4801 ggctatacat gggattactt gtgaaaataa agtattgttt actccatatt acaatcttaa
     4861 tgagattgac ttatatagca tccttagcaa aaagtatgac tttcctattc acatagaaaa
     4921 tgaagcaaac ttaacagctt tagctgagaa tactttttca actgtacata actcactact
     4981 tagtctcagt attcacagtg gatttggttc aggcattata ataaataata aattatatag
     5041 tggcagaaat ggtatgagtg gagaaattgg tcacactatc atcatgccta atggtaaact
     5101 atgtccttgt ggaaatcgtg gctgtttaga acagtattgt tcagaaaaaa aggtatttga
     5161 acagctttct tctttagaaa atattcctaa aatagattct gatatagtaa agcaactata
     5221 ttatgaagat aaccagaatg ccaaaaaagt tattcatgag ttttgtagtt acttaacaat
     5281 agctataaat aatgctatta caacttatgc tcctgaaata atatatttaa atagccaaat
     5341 aattagtgat atccctgaga tattacagat tacaaaagat atgttagtta gctcttttaa
     5401 caaaggaata aatatagaaa tttcatcatt agggtctgag gcatctctat atggtggctc
     5461 tgccgtaaat atcaagagct ttttgaatat acaaaattta actttaatta atgaaatata
     5521 agggttatga ctatatacag ccataaccct ttaaattata cgtaccataa aaataagaag
     5581 cctgcaaatg caggcttctt atttttatgc tagcttgatg cagaattcgc ccttaagtct
     5641 aaaaattagg ggagatgtaa ggatttggga aaaatagaag atgttataat cataaatatg
     5701 gtattcgtag gcttaaagtc aaaaaggagg tgaaatataa atagattttt agctaaatta
     5761 agtaagaaat aggaggagat ttattgaaca aaaaattaga aaaaccattt gtatataaga
     5821 gagagtacga tttgactgga tatgatgttg aaattttaca aaaatatgag ttagaacaag
     5881 caatatatgt ttatgttggg agtagttgtg catataacat gagagctaga agtagtaaat
     5941 ggagatacca tataagaaca aataataagt ctatatgttg taacattaaa aattttatac
     6001 ataacttgga attgttttat aaaatggaat taaagttgtc agataatatt attaatgata
     6061 agctatacta tagcaatata gcagagtttg aagaatttga aacactagaa aaagctagag
     6121 aggtagaaag tactattata agtcaatatc aatttttaga ttctataaat catatgttaa
     6181 aacaaaaaat aattttattg agtaataagg atagtgtgtt aaacataact aaaaatggaa
     6241 atacaaatta tttgaaagta aaaaataaat acatagaaaa acataagaac aagccaataa
     6301 tgagatacca tatcaactgt caattcaata cagatggaag tgtcaaaagt attacacagg
     6361 agtttgaacc aatattggaa ttaaacaaaa aaaataccct aagccgacca agcagagtat
     6421 ttttaaaata atattttaag ataacaacaa aatgagataa tactactaga caatgacaac
     6481 tcaactacca attgagttta tggagctacc aactccaata tcggtctaac tgattaagta
     6541 tctgtagtta tataataata ttgctatcaa ttttagcatc ttaacaatat tattatacat
     6601 actaagctaa aattattcaa tagttgtaaa agttgattag tcaataagta tatatttaat
     6661 gtagtgttat ctcttaaaaa aactagataa ggagataata aatatatgga acaattagat
     6721 tcaaaatata agttgaaaaa atttctaatg gcagtattta gagatggtat aggacaagga
     6781 aataatctta ttgataatga atatgttaga gtatttcaaa ataataaaag taatagtaaa
     6841 caattagaac tcggagaaga atttaaagaa tatagtaaaa caactttttt taaaaatata
     6901 gatgatatag tagaatttac cttcgcaaaa aatatttatt atgaaaatac attttttaac
     6961 ctatgtacaa ctgatggaaa agcaggaacc aatgaaaact taataaatag atatgcatta
     7021 ggatttgatt ttgacaaaaa agaattagga caaggtttta attataaaga tataattaat
     7081 ttatttacta agataggatt acattatcat atcctagttg atagtggaaa tggattccat
     7141 gtttatgtgc taattaataa aactaataac attaagttag tatcagaagt tacaaataca
     7201 ttaataaata aattgggtgc agataaacaa gcaaatttat ctactcaagt attaagagta
     7261 ccttatacat ataatattaa aaatactact aaacaagtaa aaataataca ccaagacaaa
     7321 aatatatata gatatgacat agaaaagtta gctaaaaaat attgcaaaga tgtaaaaaca
     7381 gtaggtaata ctaatacaaa atatatatta gatagtaagc taccaaattg tatagtagat
     7441 attttaaaaa atggtagtaa agatggacat aaaaacctag atttgcaaaa aatagttgtg
     7501 actttaagat tgaggaataa aagtttaagt caagtaatat ccgttgctag agaatggaac
     7561 tatatatcac aaaatagtct ttcaaatagt gagctagaat atcaagtcaa gtatatgtat
     7621 gagaagctta aaacggttaa ttttggttgt acaggttgtg agtttaatag tgattgttgg
     7681 aataaaatag aatcagattt tatatatagt gatgaagata ctttgttcaa tatgccacat
     7741 aagcactcaa aggatttgaa atataagaat aggaaagggg ttaaaataat gactggtaat
     7801 caattgttta tctataatgt gttacttaac aataaagata gagaattaaa catagacgat
     7861 ataatggagc tgataaccta taaacgtaag aagaaagtta aaaacattgt tatgagtgaa
     7921 aagacattaa gagaaacatt aaaagaactt caacataatg attatattac aaaaacaaaa
     7981 ggtgttacaa agctaggaat aaaagataca tacaatgtaa aagaagttag atgtaatata
     8041 gataaacaat atactattag ttactttgtt accatggcag taatttgggg aataatttca
     8101 actgaagaat taagattata tactcatatg agatataagc aagatttatt ggtcaaagat
     8161 gataaaataa aaggaaatat attaagaatt aatcaagagg aattagcaaa agatttagga
     8221 gtaacacagc aaagaatttc aaatatgata gaatctttat tagatactaa aattttagat
     8281 gtatgggaaa ctaaaataaa tgatagagga tttatgtact atacatatag attaaacaag
     8341 tagatttttg ataggattag aattgatttt ctagtcctat ttttatgcaa aaaaactaat
     8401 taataaaaat ttcttttggt aaaataattg tacgagaatt gcaaaaaaaa aatggcatca
     8461 aagtattgaa attaagccgt tttaaaaatt tcttttggta aaataattgt aca
//
