LOCUS       sequence-358428-        8664 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pAF256 derived plasmid encoding a xylose inducible CD16/50L 
            CBD-SmBiT and LgBiT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8664)
  AUTHORS   Phothichaisri W, Chankhamhaengdecha S, Janvilisri T, Nuadthaisong J,
            Phetruen T, Fagan RP, Chanarat S
  TITLE     Potential Role of the Host-Derived Cell-Wall Binding Domain of 
            Endolysin CD16/50L as a Molecular Anchor in Preservation of 
            Uninfected Clostridioides difficile for New Rounds of Phage 
            Infection.
  JOURNAL   Microbiol Spectr. 2022 Apr 4:e0236121. doi: 
            10.1128/spectrum.02361-21.
  PUBMED    35377223
REFERENCE   2  (bases 1 to 8664)
  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: "Microbiol 
            Spectr. 2022 Apr 4:e0236121. doi: 10.1128/spectrum.02361-21."
COMMENT     Sequence Label: pWP001
FEATURES             Location/Qualifiers
     source          1..8664
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(5958..6329)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            6362..6471
                     /note="incP origin of transfer"
     primer_bind     complement(7662..7679)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7833..8421)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7913..7932)
                     /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 ccggttatgg ctgtatatag tcataaccct
     3361 tatatttcat taattaaagt taaattttgt atattcaaaa agctcttgat atttacggca
     3421 gagccaccat atagagatgc ctcagaccct aatgatgaaa tttctatatt tattcctttg
     3481 ttaaaagagc taactaacat atcttttgta atctgtaata tctcagggat atcactaatt
     3541 atttggctat ttaaatatat tatttcagga gcataagttg taatagcatt atttatagct
     3601 attgttaagt aactacaaaa ctcatgaata acttttttgg cattctggtt atcttcataa
     3661 tatagttgct ttactatatc agaatctatt ttaggaatat tttctaaaga agaaagctgt
     3721 tcaaatacct ttttttctga acaatactgt tctaaacagc cacgatttcc acaaggacat
     3781 agtttaccat taggcatgat gatagtgtga ccaatttctc cactcatacc atttctgccg
     3841 ctatataatt tattatttat tataatgcct gaaccaaatc cactgtgaat actgagacta
     3901 agtagtgagt tatgtacagt tgaaaaagta ttctcagcta aagctgttaa gtttgcttca
     3961 ttttctatat gaataggaaa gtcatacttt ttgctaagga tgctatataa gtcaatctca
     4021 ttaagattat aatatggagt aaacaatact ttattttcac aagtaatccc atgtatagcc
     4081 aaagtaagac caattacctt ataaggagtg tctattttgg atatgttata actatttata
     4141 atttcatcta tcaattgtat aacattgtct ttacttactt gtatatctgt taatttctta
     4201 gaatttataa tagttccatc taagtaagat agagaagaaa atatatagtc gtatccaatg
     4261 tccatactta aagaaatccc tgcacattta ttaaatacta ataatatggg ttttcttccg
     4321 ccactatgag tactatttcc aattcctatt tcatgaacta gagattcatc aataagtttt
     4381 ttggtaatag cggatatagt tgctttattc aatcctatag tagaagctat acttgcccta
     4441 gaaataggac cattttttat aatttgttca agtaccaatc tttcattcat ttctcgaata
     4501 gtgtatttat cagtaaccaa tttgatattc ctccttaaaa taatattgta atacttttta
     4561 cacaaaaata aaaggttatt ttgcattgac aaagataatt aaatatttta ttattagttc
     4621 ataagttagt ttaatatact aacaaaaata aagcaagtaa aatataccta aaatataaaa
     4681 aaattaggat aggaaaacga tagttatgaa gtggcattcg agctcaggag gtacttatat
     4741 gaatagatat aaacatacaa tagtgtacag tggtgatgat aaagtatcag cagacatttt
     4801 aggactatat tataagagag aaaaagaaag ttacttagta acagatataa aagactataa
     4861 accacataga acacaaaatt tgtacgtaat tggtggagta acttgtaata aaatgaagga
     4921 aatgagtaag actacaggag aaaaatttac tcaactatat ggtaatgatg tatggtcaac
     4981 aatggataaa gctatagaat ttgtaaaaga aaaattgtct cgagggggtt ctagtggtgg
     5041 tggtggttct ggtggtggtg gttctagtgg tgttacaggt tatagacttt ttgaagaaat
     5101 tctatagctc gatcgctgca gtaaaggaga aaattttgtg tttacacttg aagattttgt
     5161 gggtgattgg gaacaaactg ctgcatataa tttagatcaa gttttagaac agggtggagt
     5221 gagttctctt ttacaaaatc ttgcagtctc agttacacca atacaaagaa tagttagaag
     5281 tggagaaaat gcattaaaga tcgatataca tgtaataatt ccttatgagg gactatcagc
     5341 agaccaaatg gcacaaattg aggaagtatt caaagttgta tatccagtag acgatcatca
     5401 ctttaaagta atattacctt atggaacttt agtaattgat ggtgtaacac caaatatgtt
     5461 aaattatttt ggtagaccat acgaagggat tgcagttttt gatggaaaga aaataaccgt
     5521 aacaggcact ttgtggaatg gaaataaaat aatagatgaa aggttaatta cacctgatgg
     5581 atctatgttg tttagagtaa ctataaacag ttagggatcc tataagtttt aataaaactt
     5641 taaatagaaa aaggcttctc tcatgagaag tcttttttat ttaaaataaa tataaaataa
     5701 aatagaggct ataaatagcc tctattttat gtgagaaatc cctaaataaa aagatgccag
     5761 tgtgctggaa ttcgccctta ggctgcgcta gggacctctt tagctccttg gaagctgtca
     5821 gtagtagggg atcggtcttg ccttgctcgt cggtgatgta cttcaccagc tccgcgaagt
     5881 cgctcttctt gatggagcgc atggggacgt gcttggcaat cacgcgcacc ccccggccgt
     5941 tttagcggct aaaaaagtca tggctctgcc ctcgggcgga ccacgcccat catgaccttg
     6001 ccaagctcgt cctgcttctc ttcgatcttc gccagcaggg cgaggatcgt ggcatcaccg
     6061 aaccgcgccg tgcgcgggtc gtcggtgagc cagagtttca gcaggccgcc caggcggccc
     6121 aggtcgccat tgatgcgggc cagctcgcgg acgtgctcat agtccacgac gcccgtgatt
     6181 ttgtagccct ggccgacggc cagcaggtag gccgacaggc tcatgccggc cgccgccgcc
     6241 ttttcctcaa tcgctcttcg ttcgtctgga aggcagtaca ccttgatagg tgggctgccc
     6301 ttcctggttg gcttggtttc atcagccatc cgcttgccct catctgttac gccggcggta
     6361 gccggccagc ctcgcagagc aggattcccg ttgagcaccg ccaggtgcga ataagggaca
     6421 gtgaagaagg aacacccgct cgcgggtggg cctacttcac ctatcctgcc cggctgacgc
     6481 cgttggatac accaaggaaa gtctacacga accctttggc aaaatcctgt atatcgtgcg
     6541 aaaaaggatg gatataccga aaaaatcgct ataatgaccc cgaagcagga agggcgaatt
     6601 ctgcagatct gaccggtctc tgaaaatata aaaaccacag attgatacta aaaccttggt
     6661 tgtgttgctt ttcggggctt aaatcaagga aaaatccttg ttttaagcct ttcaaaaaga
     6721 aacacaaggt ctttgtacta acctgtggtt atgtataaaa ttgtagattt tagggtaaca
     6781 aaaaacaccg tatttctacg atgtttttgc ttaaatactt gtttttagtt acagacaaac
     6841 ctgaagttaa ctatttatca attcctgcaa ttcgtttaca aaacggcaaa tgtgaaatcc
     6901 gtcacatact gcgtgatgaa cttgaattgc caaaggaagt ataattttgt tatcttcttt
     6961 ataatatttc cccatagtaa aaataggaat caaataatca tatcctttct gcaaattcag
     7021 attaaagcca tcgaaggttg accacggtat catagataca ttaaaaatgt tttccggagc
     7081 atttggcttt ccttccattc tatgattgtt tccataccgt tgcgtatcac tttcataatc
     7141 tgctaaaaat gatttaaagt cagacttaca ctcagtccaa aggctggaaa atgtttcagt
     7201 atcattgtga aatattgtat agcttggtat catctcatca tatatcccca attcaccatc
     7261 ttgattgatt gccgtcctaa actctgaatg gcggtttaca atcattgcaa tataataaag
     7321 cattgcagga tatagtttca ttcccttttc ctttatttgt gtgatatcca ctttaacggt
     7381 catgctgtag gtacaaggta cacttgcaaa gtagtggtca aaatactctt ttctgttcca
     7441 actattttta tcaatttttt caaataccat ctaagttccc tctcaaattc aagtttatcg
     7501 ctctaatgaa caaagatatt ataccacatt tttgtgaatt tttcaacttg cccacttcga
     7561 ctgcactccc gacttaataa cttcttgaac acttgccgaa aaagaaaaac tgccgggtac
     7621 gtacccgggg atcgatcccc ggccgagcgc tcttccgctt cctcgctcac tgactcgctg
     7681 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     7741 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     7801 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     7861 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     7921 caggcgtttc ccccttgaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     7981 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     8041 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     8101 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     8161 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     8221 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     8281 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     8341 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     8401 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     8461 tggaacgaaa actcacgtta agggattttg gtcatgaaat gcaagtttct aactaacata
     8521 catcatagtt actaaactat ggtaactata attttattaa ctatattaag cactgattag
     8581 tactataact caatataagc atatcccctg tatcgtaact agagaaccaa acgacggaaa
     8641 aagcgatata gataagttta gata
//
