LOCUS       sequence-209872-        8403 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRPF185 derived plasmid encoding an anhydrotetracyclin inducible 
            HupA-SmBiT and HupA-LgBiT fusions.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8403)
  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 8403)
  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: pAP118
FEATURES             Location/Qualifiers
     source          1..8403
                     /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(5697..6068)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            6101..6210
                     /note="incP origin of transfer"
     primer_bind     complement(7401..7418)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7572..8160)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7652..7671)
                     /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 aattttgtga ataaagctga
     4561 attagtatca aagatggcag aaaaaagtgg attaacaaag aaggaagcag aagctgcgtt
     4621 aaacgcattt atgagttctg ttcaagatgc actagtaaat aatgaaaaag ttcaattagt
     4681 tggatttgga acatttgaga caagagaaag agctgctaga caaggaagaa atccaagaga
     4741 tccagagcaa gttatagata taccagcttc taaagcacca gttttcaaag ctggaaaagg
     4801 attaaaggat ataataaatg gagcggccgc aggttctagt ggtggtggtg gttctggtgg
     4861 tggtggttct agtggtgttt ttacacttga agattttgtg ggtgattggg aacaaactgc
     4921 tgcatataat ttagatcaag ttttagaaca gggtggagtg agttctcttt tacaaaatct
     4981 tgcagtctca gttacaccaa tacaaagaat agttagaagt ggagaaaatg cattaaagat
     5041 cgatatacat gtaataattc cttatgaggg actatcagca gaccaaatgg cacaaattga
     5101 ggaagtattc aaagttgtat atccagtaga cgatcatcac tttaaagtaa tattacctta
     5161 tggaacttta gtaattgatg gtgtaacacc aaatatgtta aattattttg gtagaccata
     5221 cgaagggatt gcagtttttg atggaaagaa aataaccgta acaggcactt tgtggaatgg
     5281 aaataaaata atagatgaaa ggttaattac acctgatgga tctatgttgt ttagagtaac
     5341 tataaacagt tagggatcct ataagtttta ataaaacttt aaatagaaaa aggcttctct
     5401 catgagaagt cttttttatt taaaataaat ataaaataaa atagaggcta taaatagcct
     5461 ctattttatg tgagaaatcc ctaaataaaa agatgccagt gtgctggaat tcgcccttag
     5521 gctgcgctag ggacctcttt agctccttgg aagctgtcag tagtagggga tcggtcttgc
     5581 cttgctcgtc ggtgatgtac ttcaccagct ccgcgaagtc gctcttcttg atggagcgca
     5641 tggggacgtg cttggcaatc acgcgcaccc cccggccgtt ttagcggcta aaaaagtcat
     5701 ggctctgccc tcgggcggac cacgcccatc atgaccttgc caagctcgtc ctgcttctct
     5761 tcgatcttcg ccagcagggc gaggatcgtg gcatcaccga accgcgccgt gcgcgggtcg
     5821 tcggtgagcc agagtttcag caggccgccc aggcggccca ggtcgccatt gatgcgggcc
     5881 agctcgcgga cgtgctcata gtccacgacg cccgtgattt tgtagccctg gccgacggcc
     5941 agcaggtagg ccgacaggct catgccggcc gccgccgcct tttcctcaat cgctcttcgt
     6001 tcgtctggaa ggcagtacac cttgataggt gggctgccct tcctggttgg cttggtttca
     6061 tcagccatcc gcttgccctc atctgttacg ccggcggtag ccggccagcc tcgcagagca
     6121 ggattcccgt tgagcaccgc caggtgcgaa taagggacag tgaagaagga acacccgctc
     6181 gcgggtgggc ctacttcacc tatcctgccc ggctgacgcc gttggataca ccaaggaaag
     6241 tctacacgaa ccctttggca aaatcctgta tatcgtgcga aaaaggatgg atataccgaa
     6301 aaaatcgcta taatgacccc gaagcaggaa gggcgaattc tgcagatctg accggtctct
     6361 gaaaatataa aaaccacaga ttgatactaa aaccttggtt gtgttgcttt tcggggctta
     6421 aatcaaggaa aaatccttgt tttaagcctt tcaaaaagaa acacaaggtc tttgtactaa
     6481 cctgtggtta tgtataaaat tgtagatttt agggtaacaa aaaacaccgt atttctacga
     6541 tgtttttgct taaatacttg tttttagtta cagacaaacc tgaagttaac tatttatcaa
     6601 ttcctgcaat tcgtttacaa aacggcaaat gtgaaatccg tcacatactg cgtgatgaac
     6661 ttgaattgcc aaaggaagta taattttgtt atcttcttta taatatttcc ccatagtaaa
     6721 aataggaatc aaataatcat atcctttctg caaattcaga ttaaagccat cgaaggttga
     6781 ccacggtatc atagatacat taaaaatgtt ttccggagca tttggctttc cttccattct
     6841 atgattgttt ccataccgtt gcgtatcact ttcataatct gctaaaaatg atttaaagtc
     6901 agacttacac tcagtccaaa ggctggaaaa tgtttcagta tcattgtgaa atattgtata
     6961 gcttggtatc atctcatcat atatccccaa ttcaccatct tgattgattg ccgtcctaaa
     7021 ctctgaatgg cggtttacaa tcattgcaat ataataaagc attgcaggat atagtttcat
     7081 tcccttttcc tttatttgtg tgatatccac tttaacggtc atgctgtagg tacaaggtac
     7141 acttgcaaag tagtggtcaa aatactcttt tctgttccaa ctatttttat caattttttc
     7201 aaataccatc taagttccct ctcaaattca agtttatcgc tctaatgaac aaagatatta
     7261 taccacattt ttgtgaattt ttcaacttgc ccacttcgac tgcactcccg acttaataac
     7321 ttcttgaaca cttgccgaaa aagaaaaact gccgggtacg tacccgggga tcgatccccg
     7381 gccgagcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     7441 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     7501 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     7561 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     7621 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc cccttgaagc
     7681 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     7741 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     7801 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     7861 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     7921 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     7981 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     8041 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     8101 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     8161 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     8221 gggattttgg tcatgaaatg caagtttcta actaacatac atcatagtta ctaaactatg
     8281 gtaactataa ttttattaac tatattaagc actgattagt actataactc aatataagca
     8341 tatcccctgt atcgtaacta gagaaccaaa cgacggaaaa agcgatatag ataagtttag
     8401 ata
//
