LOCUS       sequence-209909-        8073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRPF185 derived plasmid encoding an anhydrotetracyclin inducible 
            HupA-SmBiT and LgBiT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8073)
  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 8073)
  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: pAF256
FEATURES             Location/Qualifiers
     source          1..8073
                     /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(5367..5738)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            5771..5880
                     /note="incP origin of transfer"
     primer_bind     complement(7071..7088)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7242..7830)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7322..7341)
                     /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 gtgaataaag ctgaattagt atcaaagatg gcagaaaaaa gtggattaac
     4201 aaagaaggaa gcagaagctg cgttaaacgc atttatgagt tctgttcaag atgcactagt
     4261 aaataatgaa aaagttcaat tagttggatt tggaacattt gagacaagag aaagagctgc
     4321 tagacaagga agaaatccaa gagatccaga gcaagttata gatataccag cttctaaagc
     4381 accagttttc aaagctggaa aaggattaaa ggatataata aatggatctc gagggggttc
     4441 tagtggtggt ggtggttctg gtggtggtgg ttctagtggt gttacaggtt atagactttt
     4501 tgaagaaatt ctatagctcg atcgctgcag taaaggagaa aattttgtgt ttacacttga
     4561 agattttgtg ggtgattggg aacaaactgc tgcatataat ttagatcaag ttttagaaca
     4621 gggtggagtg agttctcttt tacaaaatct tgcagtctca gttacaccaa tacaaagaat
     4681 agttagaagt ggagaaaatg cattaaagat cgatatacat gtaataattc cttatgaggg
     4741 actatcagca gaccaaatgg cacaaattga ggaagtattc aaagttgtat atccagtaga
     4801 cgatcatcac tttaaagtaa tattacctta tggaacttta gtaattgatg gtgtaacacc
     4861 aaatatgtta aattattttg gtagaccata cgaagggatt gcagtttttg atggaaagaa
     4921 aataaccgta acaggcactt tgtggaatgg aaataaaata atagatgaaa ggttaattac
     4981 acctgatgga tctatgttgt ttagagtaac tataaacagt tagggatcct ataagtttta
     5041 ataaaacttt aaatagaaaa aggcttctct catgagaagt cttttttatt taaaataaat
     5101 ataaaataaa atagaggcta taaatagcct ctattttatg tgagaaatcc ctaaataaaa
     5161 agatgccagt gtgctggaat tcgcccttag gctgcgctag ggacctcttt agctccttgg
     5221 aagctgtcag tagtagggga tcggtcttgc cttgctcgtc ggtgatgtac ttcaccagct
     5281 ccgcgaagtc gctcttcttg atggagcgca tggggacgtg cttggcaatc acgcgcaccc
     5341 cccggccgtt ttagcggcta aaaaagtcat ggctctgccc tcgggcggac cacgcccatc
     5401 atgaccttgc caagctcgtc ctgcttctct tcgatcttcg ccagcagggc gaggatcgtg
     5461 gcatcaccga accgcgccgt gcgcgggtcg tcggtgagcc agagtttcag caggccgccc
     5521 aggcggccca ggtcgccatt gatgcgggcc agctcgcgga cgtgctcata gtccacgacg
     5581 cccgtgattt tgtagccctg gccgacggcc agcaggtagg ccgacaggct catgccggcc
     5641 gccgccgcct tttcctcaat cgctcttcgt tcgtctggaa ggcagtacac cttgataggt
     5701 gggctgccct tcctggttgg cttggtttca tcagccatcc gcttgccctc atctgttacg
     5761 ccggcggtag ccggccagcc tcgcagagca ggattcccgt tgagcaccgc caggtgcgaa
     5821 taagggacag tgaagaagga acacccgctc gcgggtgggc ctacttcacc tatcctgccc
     5881 ggctgacgcc gttggataca ccaaggaaag tctacacgaa ccctttggca aaatcctgta
     5941 tatcgtgcga aaaaggatgg atataccgaa aaaatcgcta taatgacccc gaagcaggaa
     6001 gggcgaattc tgcagatctg accggtctct gaaaatataa aaaccacaga ttgatactaa
     6061 aaccttggtt gtgttgcttt tcggggctta aatcaaggaa aaatccttgt tttaagcctt
     6121 tcaaaaagaa acacaaggtc tttgtactaa cctgtggtta tgtataaaat tgtagatttt
     6181 agggtaacaa aaaacaccgt atttctacga tgtttttgct taaatacttg tttttagtta
     6241 cagacaaacc tgaagttaac tatttatcaa ttcctgcaat tcgtttacaa aacggcaaat
     6301 gtgaaatccg tcacatactg cgtgatgaac ttgaattgcc aaaggaagta taattttgtt
     6361 atcttcttta taatatttcc ccatagtaaa aataggaatc aaataatcat atcctttctg
     6421 caaattcaga ttaaagccat cgaaggttga ccacggtatc atagatacat taaaaatgtt
     6481 ttccggagca tttggctttc cttccattct atgattgttt ccataccgtt gcgtatcact
     6541 ttcataatct gctaaaaatg atttaaagtc agacttacac tcagtccaaa ggctggaaaa
     6601 tgtttcagta tcattgtgaa atattgtata gcttggtatc atctcatcat atatccccaa
     6661 ttcaccatct tgattgattg ccgtcctaaa ctctgaatgg cggtttacaa tcattgcaat
     6721 ataataaagc attgcaggat atagtttcat tcccttttcc tttatttgtg tgatatccac
     6781 tttaacggtc atgctgtagg tacaaggtac acttgcaaag tagtggtcaa aatactcttt
     6841 tctgttccaa ctatttttat caattttttc aaataccatc taagttccct ctcaaattca
     6901 agtttatcgc tctaatgaac aaagatatta taccacattt ttgtgaattt ttcaacttgc
     6961 ccacttcgac tgcactcccg acttaataac ttcttgaaca cttgccgaaa aagaaaaact
     7021 gccgggtacg tacccgggga tcgatccccg gccgagcgct cttccgcttc ctcgctcact
     7081 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     7141 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     7201 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     7261 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     7321 taaagatacc aggcgtttcc cccttgaagc tccctcgtgc gctctcctgt tccgaccctg
     7381 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     7441 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     7501 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     7561 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     7621 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     7681 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     7741 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     7801 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     7861 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgaaatg caagtttcta
     7921 actaacatac atcatagtta ctaaactatg gtaactataa ttttattaac tatattaagc
     7981 actgattagt actataactc aatataagca tatcccctgt atcgtaacta gagaaccaaa
     8041 cgacggaaaa agcgatatag ataagtttag ata
//
