LOCUS       sequence-209908-        7710 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRPF185 derived plasmid encoding an anhydrotetracyclin inducible 
            BitlucOpt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7710)
  AUTHORS   Oliveira Paiva AM, Friggen AH, Qin L, Douwes R, Dame RT, Smits WK
  TITLE     The Bacterial Chromatin Protein HupA Can Remodel DNA and Associates 
            with the Nucleoid in Clostridium difficile.
  JOURNAL   J Mol Biol. 2019 Jan 8. pii: S0022-2836(18)31160-4. doi: 
            10.1016/j.jmb.2019.01.001.
  PUBMED    30633871
REFERENCE   2  (bases 1 to 7710)
  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 Mol Biol.
            2019 Jan 8. pii: S0022-2836(18)31160-4. doi: 
            10.1016/j.jmb.2019.01.001."
COMMENT     Sequence Label: pAF259
FEATURES             Location/Qualifiers
     source          1..7710
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(3333..3959)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     3864..3883
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     protein_bind    4066..4084
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     CDS             complement(5004..5375)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            5408..5517
                     /note="incP origin of transfer"
     primer_bind     complement(6708..6725)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6879..7467)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6959..6978)
                     /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 ccttaagacc cactttcaca tttaagttgt
     3361 ttttctaatc cgcatatgat caattcaagg ccgaataaga aggctggctc tgcaccttgg
     3421 tgatcaaata attcgatagc ttgtcgtaat aatggcggca tactatcagt agtaggtgtt
     3481 tccctttctt ctttagcgac ttgatgctct tgatcttcca atacgcaacc taaagtaaaa
     3541 tgccccacag cgctgagtgc atataatgca ttctctagtg aaaaaccttg ttggcataaa
     3601 aaggctaatt gattttcgag agtttcatac tgtttttctg taggccgtgt acctaaatgt
     3661 acttttgctc catcgcgatg acttagtaaa gcacatctaa aacttttagc gttattacgt
     3721 aaaaaatctt gccagctttc cccttctaaa gggcaaaagt gagtatggtg cctatctaac
     3781 atctcaatgg ctaaggcgtc gagcaaagcc cgcttatttt ttacatgcca atacaatgta
     3841 ggctgctcta cacctagctt ctgggcgagt ttacgggttg ttaaaccttc gattccgacc
     3901 tcattaagca gctctaatgc gctgttaatc actttacttt tatctaatct agacatcatt
     3961 aattcctcct ttttgttgac attatatcat tgatagagtt atttgtcaaa ctagtttttt
     4021 atttcgatgc cctggacttc atgaaaaact aaaaaaaata ttgacactct atcattgata
     4081 gagtataatt aaaataagct tgatcgtagc gttaacagat ctgagctcct gcagtaaagg
     4141 agaaaatttt gtgtttacac ttgaagattt tgtgggtgat tgggaacaaa ctgctgcata
     4201 taatttagat caagttttag aacagggtgg agtgagttct cttttacaaa atcttgcagt
     4261 ctcagttaca ccaatacaaa gaatagttag aagtggagaa aatgcattaa agatcgatat
     4321 acatgtaata attccttatg agggactatc agcagaccaa atggcacaaa ttgaggaagt
     4381 attcaaagtt gtatatccag tagacgatca tcactttaaa gtaatattac cttatggaac
     4441 tttagtaatt gatggtgtaa caccaaatat gttaaattat tttggtagac catacgaagg
     4501 gattgcagtt tttgatggaa agaaaataac cgtaacaggc actttgtgga atggaaataa
     4561 aataatagat gaaaggttaa ttacacctga tggatctatg ttgtttagag taactataaa
     4621 cagtgttaca ggttatagac tttttgaaga aattctatag ggatcctata agttttaata
     4681 aaactttaaa tagaaaaagg cttctctcat gagaagtctt ttttatttaa aataaatata
     4741 aaataaaata gaggctataa atagcctcta ttttatgtga gaaatcccta aataaaaaga
     4801 tgccagtgtg ctggaattcg cccttaggct gcgctaggga cctctttagc tccttggaag
     4861 ctgtcagtag taggggatcg gtcttgcctt gctcgtcggt gatgtacttc accagctccg
     4921 cgaagtcgct cttcttgatg gagcgcatgg ggacgtgctt ggcaatcacg cgcacccccc
     4981 ggccgtttta gcggctaaaa aagtcatggc tctgccctcg ggcggaccac gcccatcatg
     5041 accttgccaa gctcgtcctg cttctcttcg atcttcgcca gcagggcgag gatcgtggca
     5101 tcaccgaacc gcgccgtgcg cgggtcgtcg gtgagccaga gtttcagcag gccgcccagg
     5161 cggcccaggt cgccattgat gcgggccagc tcgcggacgt gctcatagtc cacgacgccc
     5221 gtgattttgt agccctggcc gacggccagc aggtaggccg acaggctcat gccggccgcc
     5281 gccgcctttt cctcaatcgc tcttcgttcg tctggaaggc agtacacctt gataggtggg
     5341 ctgcccttcc tggttggctt ggtttcatca gccatccgct tgccctcatc tgttacgccg
     5401 gcggtagccg gccagcctcg cagagcagga ttcccgttga gcaccgccag gtgcgaataa
     5461 gggacagtga agaaggaaca cccgctcgcg ggtgggccta cttcacctat cctgcccggc
     5521 tgacgccgtt ggatacacca aggaaagtct acacgaaccc tttggcaaaa tcctgtatat
     5581 cgtgcgaaaa aggatggata taccgaaaaa atcgctataa tgaccccgaa gcaggaaggg
     5641 cgaattctgc agatctgacc ggtctctgaa aatataaaaa ccacagattg atactaaaac
     5701 cttggttgtg ttgcttttcg gggcttaaat caaggaaaaa tccttgtttt aagcctttca
     5761 aaaagaaaca caaggtcttt gtactaacct gtggttatgt ataaaattgt agattttagg
     5821 gtaacaaaaa acaccgtatt tctacgatgt ttttgcttaa atacttgttt ttagttacag
     5881 acaaacctga agttaactat ttatcaattc ctgcaattcg tttacaaaac ggcaaatgtg
     5941 aaatccgtca catactgcgt gatgaacttg aattgccaaa ggaagtataa ttttgttatc
     6001 ttctttataa tatttcccca tagtaaaaat aggaatcaaa taatcatatc ctttctgcaa
     6061 attcagatta aagccatcga aggttgacca cggtatcata gatacattaa aaatgttttc
     6121 cggagcattt ggctttcctt ccattctatg attgtttcca taccgttgcg tatcactttc
     6181 ataatctgct aaaaatgatt taaagtcaga cttacactca gtccaaaggc tggaaaatgt
     6241 ttcagtatca ttgtgaaata ttgtatagct tggtatcatc tcatcatata tccccaattc
     6301 accatcttga ttgattgccg tcctaaactc tgaatggcgg tttacaatca ttgcaatata
     6361 ataaagcatt gcaggatata gtttcattcc cttttccttt atttgtgtga tatccacttt
     6421 aacggtcatg ctgtaggtac aaggtacact tgcaaagtag tggtcaaaat actcttttct
     6481 gttccaacta tttttatcaa ttttttcaaa taccatctaa gttccctctc aaattcaagt
     6541 ttatcgctct aatgaacaaa gatattatac cacatttttg tgaatttttc aacttgccca
     6601 cttcgactgc actcccgact taataacttc ttgaacactt gccgaaaaag aaaaactgcc
     6661 gggtacgtac ccggggatcg atccccggcc gagcgctctt ccgcttcctc gctcactgac
     6721 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     6781 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6841 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6901 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6961 agataccagg cgtttccccc ttgaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     7021 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     7081 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     7141 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     7201 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     7261 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     7321 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     7381 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     7441 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     7501 gctcagtgga acgaaaactc acgttaaggg attttggtca tgaaatgcaa gtttctaact
     7561 aacatacatc atagttacta aactatggta actataattt tattaactat attaagcact
     7621 gattagtact ataactcaat ataagcatat cccctgtatc gtaactagag aaccaaacga
     7681 cggaaaaagc gatatagata agtttagata
//
