LOCUS       sequence-274045-        7968 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Extracellular control for HiBiTopt experiments.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7968)
  AUTHORS   Oliveira Paiva AM, de Jong L, Friggen AH, Smits WK, Corver J
  TITLE     The C-terminal domain of Clostridioides difficile TcdC is exposed on
            the bacterial cell surface.
  JOURNAL   J Bacteriol. 2020 Aug 31. pii: JB.00771-19. doi: 
            10.1128/JB.00771-19.
  PUBMED    32868401
REFERENCE   2  (bases 1 to 7968)
  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 
            Bacteriol. 2020 Aug 31. pii: JB.00771-19. doi: 10.1128/JB.00771-19."
COMMENT     Sequence Label: pAP233
FEATURES             Location/Qualifiers
     source          1..7968
                     /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(5262..5633)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            5666..5775
                     /note="incP origin of transfer"
     primer_bind     complement(6966..6983)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7137..7725)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7217..7236)
                     /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 atgttgaaaa aattatatag aatagtaatt aacattatac tagtattagt
     4201 aatactttac agtggattta atatatactc aaaacttacc aaatacaatc atgatactaa
     4261 aatatcctca gaacttcaaa aaaaagaata taaaaaagag gatttatcaa agattaacag
     4321 tgattttaaa ttttggcttt ctgttgagaa tacaaatata aattatccag ttgtacaatc
     4381 taaagataat tcttactatc ttgacaaaga tttttataaa aaagattcta tttctggtac
     4441 actgtttatg gattatagaa ataaatctat tgatgacaaa aatatcataa tatatggaca
     4501 taatatgaaa aataaaacta tgttcaacaa cctaaataag tttaaagatg ctgacttctt
     4561 taaaaaaaat aataaaataa aaataacctt gaatggaaaa gaatttttat atgatgtttt
     4621 ttctgcttat attgtagaat ctgattatga ttatctaaaa acaaatttta ataatgaatc
     4681 tgattatcaa aattacataa atgatattac ttctaaatca ttgtataaat ctcctattaa
     4741 ggtaaatagc aatgataaaa tagttacact ttctacatgt acctatgaat ttgatgatgc
     4801 cagaatggtg attcatggta gattgatttc tcgagggggt tctagtggtg gtggtggttc
     4861 tggtggtggt ggttctagtg gtgttagtgg ttggagactt tttaagaaaa tttcttaggg
     4921 atcctataag ttttaataaa actttaaata gaaaaaggct tctctcatga gaagtctttt
     4981 ttatttaaaa taaatataaa ataaaataga ggctataaat agcctctatt ttatgtgaga
     5041 aatccctaaa taaaaagatg ccagtgtgct ggaattcgcc cttaggctgc gctagggacc
     5101 tctttagctc cttggaagct gtcagtagta ggggatcggt cttgccttgc tcgtcggtga
     5161 tgtacttcac cagctccgcg aagtcgctct tcttgatgga gcgcatgggg acgtgcttgg
     5221 caatcacgcg caccccccgg ccgttttagc ggctaaaaaa gtcatggctc tgccctcggg
     5281 cggaccacgc ccatcatgac cttgccaagc tcgtcctgct tctcttcgat cttcgccagc
     5341 agggcgagga tcgtggcatc accgaaccgc gccgtgcgcg ggtcgtcggt gagccagagt
     5401 ttcagcaggc cgcccaggcg gcccaggtcg ccattgatgc gggccagctc gcggacgtgc
     5461 tcatagtcca cgacgcccgt gattttgtag ccctggccga cggccagcag gtaggccgac
     5521 aggctcatgc cggccgccgc cgccttttcc tcaatcgctc ttcgttcgtc tggaaggcag
     5581 tacaccttga taggtgggct gcccttcctg gttggcttgg tttcatcagc catccgcttg
     5641 ccctcatctg ttacgccggc ggtagccggc cagcctcgca gagcaggatt cccgttgagc
     5701 accgccaggt gcgaataagg gacagtgaag aaggaacacc cgctcgcggg tgggcctact
     5761 tcacctatcc tgcccggctg acgccgttgg atacaccaag gaaagtctac acgaaccctt
     5821 tggcaaaatc ctgtatatcg tgcgaaaaag gatggatata ccgaaaaaat cgctataatg
     5881 accccgaagc aggaagggcg aattctgcag atctgaccgg tctctgaaaa tataaaaacc
     5941 acagattgat actaaaacct tggttgtgtt gcttttcggg gcttaaatca aggaaaaatc
     6001 cttgttttaa gcctttcaaa aagaaacaca aggtctttgt actaacctgt ggttatgtat
     6061 aaaattgtag attttagggt aacaaaaaac accgtatttc tacgatgttt ttgcttaaat
     6121 acttgttttt agttacagac aaacctgaag ttaactattt atcaattcct gcaattcgtt
     6181 tacaaaacgg caaatgtgaa atccgtcaca tactgcgtga tgaacttgaa ttgccaaagg
     6241 aagtataatt ttgttatctt ctttataata tttccccata gtaaaaatag gaatcaaata
     6301 atcatatcct ttctgcaaat tcagattaaa gccatcgaag gttgaccacg gtatcataga
     6361 tacattaaaa atgttttccg gagcatttgg ctttccttcc attctatgat tgtttccata
     6421 ccgttgcgta tcactttcat aatctgctaa aaatgattta aagtcagact tacactcagt
     6481 ccaaaggctg gaaaatgttt cagtatcatt gtgaaatatt gtatagcttg gtatcatctc
     6541 atcatatatc cccaattcac catcttgatt gattgccgtc ctaaactctg aatggcggtt
     6601 tacaatcatt gcaatataat aaagcattgc aggatatagt ttcattccct tttcctttat
     6661 ttgtgtgata tccactttaa cggtcatgct gtaggtacaa ggtacacttg caaagtagtg
     6721 gtcaaaatac tcttttctgt tccaactatt tttatcaatt ttttcaaata ccatctaagt
     6781 tccctctcaa attcaagttt atcgctctaa tgaacaaaga tattatacca catttttgtg
     6841 aatttttcaa cttgcccact tcgactgcac tcccgactta ataacttctt gaacacttgc
     6901 cgaaaaagaa aaactgccgg gtacgtaccc ggggatcgat ccccggccga gcgctcttcc
     6961 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     7021 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     7081 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     7141 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     7201 aacccgacag gactataaag ataccaggcg tttccccctt gaagctccct cgtgcgctct
     7261 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     7321 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     7381 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     7441 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     7501 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     7561 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     7621 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     7681 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     7741 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     7801 aaatgcaagt ttctaactaa catacatcat agttactaaa ctatggtaac tataatttta
     7861 ttaactatat taagcactga ttagtactat aactcaatat aagcatatcc cctgtatcgt
     7921 aactagagaa ccaaacgacg gaaaaagcga tatagataag tttagata
//
