LOCUS       sequence-235309-        7885 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli-C. difficile shuttle vector: tetracycline-inducible promoter
            (Ptet) driving expression of cyan fluorescent protein (CfpOpt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7885)
  AUTHORS   Ransom EM, Williams KB, Weiss DS, Ellermeier CD
  TITLE     Identification and characterization of a gene cluster required for 
            proper rod shape, cell division, and pathogenesis in Clostridium 
            difficile.
  JOURNAL   J Bacteriol. 2014 Jun;196(12):2290-300. doi: 10.1128/JB.00038-14. 
            Epub 2014 Apr 11.
  PUBMED    24727226
REFERENCE   2  (bases 1 to 7885)
  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"; doi: 
            "10.1128/JB.00038-14"; journalName: "J Bacteriol"; date: "2014-06"; 
            volume: "196"; issue: "12"; pages: "2290-300"
COMMENT     Sequence Label: pRAN334
FEATURES             Location/Qualifiers
     source          1..7885
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      611..1199
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1100..1119
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1353..1370
                     /label=L4440
                     /note="L4440 vector, forward primer"
     oriT            2561..2670
                     /note="incP origin of transfer"
     CDS             2703..3074
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     protein_bind    4204..4222
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     CDS             4329..4955
                     /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     complement(4405..4424)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
ORIGIN
        1 tcttttggta aaataattgt acatatatat gtagtatata tatatatgtt ttttagagaa
       61 tgtataacta gaatatagag ctagaatata gagaatgtat aactagaata tagagctaga
      121 atatagagaa tgtataacta gaatatagag ctagaatata gagaatgtat aactagaata
      181 tagagctaga atatagagaa tgtataacta gaatatagag ctagaatata gagaatgtat
      241 aactagaata tagagctaga atcctaatta gtaggtgctt ttttaaaaca agttaaaaat
      301 caaaaatagt attagtaagc attggaaatg ctagattcta aaatagaaaa gtaaaaaatt
      361 ggtgcactat ctaaacttat ctatatcgct ttttccgtcg tttggttctc tagttacgat
      421 acaggggata tgcttatatt gagttatagt actaatcagt gcttaatata gttaataaaa
      481 ttatagttac catagtttag taactatgat gtatgttagt tagaaacttg catttcatga
      541 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
      601 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
      661 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
      721 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
      781 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
      841 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
      901 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
      961 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     1021 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     1081 gcacgaggga gcttcaaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     1141 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     1201 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     1261 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     1321 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     1381 agcgctcggc cggggatcga tccccgggta cgtacccggc agtttttctt tttcggcaag
     1441 tgttcaagaa gttattaagt cgggagtgca gtcgaagtgg gcaagttgaa aaattcacaa
     1501 aaatgtggta taatatcttt gttcattaga gcgataaact tgaatttgag agggaactta
     1561 gatggtattt gaaaaaattg ataaaaatag ttggaacaga aaagagtatt ttgaccacta
     1621 ctttgcaagt gtaccttgta cctacagcat gaccgttaaa gtggatatca cacaaataaa
     1681 ggaaaaggga atgaaactat atcctgcaat gctttattat attgcaatga ttgtaaaccg
     1741 ccattcagag tttaggacgg caatcaatca agatggtgaa ttggggatat atgatgagat
     1801 gataccaagc tatacaatat ttcacaatga tactgaaaca ttttccagcc tttggactga
     1861 gtgtaagtct gactttaaat catttttagc agattatgaa agtgatacgc aacggtatgg
     1921 aaacaatcat agaatggaag gaaagccaaa tgctccggaa aacattttta atgtatctat
     1981 gataccgtgg tcaaccttcg atggctttaa tctgaatttg cagaaaggat atgattattt
     2041 gattcctatt tttactatgg ggaaatatta taaagaagat aacaaaatta tacttccttt
     2101 ggcaattcaa gttcatcacg cagtatgtga cggatttcac atttgccgtt ttgtaaacga
     2161 attgcaggaa ttgataaata gttaacttca ggtttgtctg taactaaaaa caagtattta
     2221 agcaaaaaca tcgtagaaat acggtgtttt ttgttaccct aaaatctaca attttataca
     2281 taaccacagg ttagtacaaa gaccttgtgt ttctttttga aaggcttaaa acaaggattt
     2341 ttccttgatt taagccccga aaagcaacac aaccaaggtt ttagtatcaa tctgtggttt
     2401 ttatattttc agagaccggt cagatctgca gaattcgccc ttcctgcttc ggggtcatta
     2461 tagcgatttt ttcggtatat ccatcctttt tcgcacgata tacaggattt tgccaaaggg
     2521 ttcgtgtaga ctttccttgg tgtatccaac ggcgtcagcc gggcaggata ggtgaagtag
     2581 gcccacccgc gagcgggtgt tccttcttca ctgtccctta ttcgcacctg gcggtgctca
     2641 acgggaatcc tgctctgcga ggctggccgg ctaccgccgg cgtaacagat gagggcaagc
     2701 ggatggctga tgaaaccaag ccaaccagga agggcagccc acctatcaag gtgtactgcc
     2761 ttccagacga acgaagagcg attgaggaaa aggcggcggc ggccggcatg agcctgtcgg
     2821 cctacctgct ggccgtcggc cagggctaca aaatcacggg cgtcgtggac tatgagcacg
     2881 tccgcgagct ggcccgcatc aatggcgacc tgggccgcct gggcggcctg ctgaaactct
     2941 ggctcaccga cgacccgcgc acggcgcggt tcggtgatgc cacgatcctc gccctgctgg
     3001 cgaagatcga agagaagcag gacgagcttg gcaaggtcat gatgggcgtg gtccgcccga
     3061 gggcagagcc atgacttttt tagccgctaa aacggccggg gggtgcgcgt gattgccaag
     3121 cacgtcccca tgcgctccat caagaagagc gacttcgcgg agctggtgaa gtacatcacc
     3181 gacgagcaag gcaagaccga tcccctacta ctgacagctt ccaaggagct aaagaggtcc
     3241 ctagcgcagc ctaagggcga attccagcac actggcatct ttttatttag ggatttctca
     3301 cataaaatag aggctattta tagcctctat tttattttat atttatttta aataaaaaag
     3361 acttctcatg agagaagcct ttttctattt aaagttttat taaaacttat aggatcctta
     3421 cttatataat tcatccattc ctagtgtaat tcctgctgct gttacgaatt ctaataatac
     3481 catgtggtct cttttttcat ttgggtcttt agataaagcg ctttgcgtgc ttaagtaatg
     3541 attatctggt agaagcactg ggccatctcc gataggtgta ttttgttgat agtgatctgc
     3601 taattgtact gatccgtctt ctatattgtg tcttatttta aagttagctt taattccatt
     3661 tttctgttta tctgccgtta tgtatacgtt atgactgatg taattgtatt ctaatttatg
     3721 tcctaggata tttccatctt ctttaaagtc tattcctttt aattcaattc tgtttactaa
     3781 tgtatctcct tcaaatttta cttctgctct cgttttatag ttaccatcat ctttaaaaaa
     3841 tattgttctt tcttgtacgt atccttcagg cattgctgat ttaaaaaagt cgtgttgttt
     3901 catatgatct ggatatcttg aaaaacattg tactccccat gttaaagtag tcacaagtgt
     3961 gggccatgga accggtagtt tacctgttgt acaaatgaat ttaagtgtta attttccgta
     4021 tgttgcatct ccttctcctt ctcctgatac tgaaaatttg tgtccattta catctccgtc
     4081 aagttctact aaaattggta ctacccctgt aaataattct tctccttttg aaaccataaa
     4141 attttctcct ttactgcggg agctcagatc tgttaacgct acgatcaagc ttattttaat
     4201 tatactctat caatgataga gtgtcaatat tttttttagt ttttcatgaa gtccagggca
     4261 tcgaaataaa aaactagttt gacaaataac tctatcaatg atataatgtc aacaaaaagg
     4321 aggaattaat gatgtctaga ttagataaaa gtaaagtgat taacagcgca ttagagctgc
     4381 ttaatgaggt cggaatcgaa ggtttaacaa cccgtaaact cgcccagaag ctaggtgtag
     4441 agcagcctac attgtattgg catgtaaaaa ataagcgggc tttgctcgac gccttagcca
     4501 ttgagatgtt agataggcac catactcact tttgcccttt agaaggggaa agctggcaag
     4561 attttttacg taataacgct aaaagtttta gatgtgcttt actaagtcat cgcgatggag
     4621 caaaagtaca tttaggtaca cggcctacag aaaaacagta tgaaactctc gaaaatcaat
     4681 tagccttttt atgccaacaa ggtttttcac tagagaatgc attatatgca ctcagcgctg
     4741 tggggcattt tactttaggt tgcgtattgg aagatcaaga gcatcaagtc gctaaagaag
     4801 aaagggaaac acctactact gatagtatgc cgccattatt acgacaagct atcgaattat
     4861 ttgatcacca aggtgcagag ccagccttct tattcggcct tgaattgatc atatgcggat
     4921 tagaaaaaca acttaaatgt gaaagtgggt cttaaggtac cataaaaata agaagcctgc
     4981 aaatgcaggc ttcttatttt tatgctagct tgatgcagaa ttcgccctta agtctaaaaa
     5041 ttaggggaga tgtaaggatt tgggaaaaat agaagatgtt ataatcataa atatggtatt
     5101 cgtaggctta aagtcaaaaa ggaggtgaaa tataaataga tttttagcta aattaagtaa
     5161 gaaataggag gagatttatt gaacaaaaaa ttagaaaaac catttgtata taagagagag
     5221 tacgatttga ctggatatga tgttgaaatt ttacaaaaat atgagttaga acaagcaata
     5281 tatgtttatg ttgggagtag ttgtgcatat aacatgagag ctagaagtag taaatggaga
     5341 taccatataa gaacaaataa taagtctata tgttgtaaca ttaaaaattt tatacataac
     5401 ttggaattgt tttataaaat ggaattaaag ttgtcagata atattattaa tgataagcta
     5461 tactatagca atatagcaga gtttgaagaa tttgaaacac tagaaaaagc tagagaggta
     5521 gaaagtacta ttataagtca atatcaattt ttagattcta taaatcatat gttaaaacaa
     5581 aaaataattt tattgagtaa taaggatagt gtgttaaaca taactaaaaa tggaaataca
     5641 aattatttga aagtaaaaaa taaatacata gaaaaacata agaacaagcc aataatgaga
     5701 taccatatca actgtcaatt caatacagat ggaagtgtca aaagtattac acaggagttt
     5761 gaaccaatat tggaattaaa caaaaaaaat accctaagcc gaccaagcag agtattttta
     5821 aaataatatt ttaagataac aacaaaatga gataatacta ctagacaatg acaactcaac
     5881 taccaattga gtttatggag ctaccaactc caatatcggt ctaactgatt aagtatctgt
     5941 agttatataa taatattgct atcaatttta gcatcttaac aatattatta tacatactaa
     6001 gctaaaatta ttcaatagtt gtaaaagttg attagtcaat aagtatatat ttaatgtagt
     6061 gttatctctt aaaaaaacta gataaggaga taataaatat atggaacaat tagattcaaa
     6121 atataagttg aaaaaatttc taatggcagt atttagagat ggtataggac aaggaaataa
     6181 tcttattgat aatgaatatg ttagagtatt tcaaaataat aaaagtaata gtaaacaatt
     6241 agaactcgga gaagaattta aagaatatag taaaacaact ttttttaaaa atatagatga
     6301 tatagtagaa tttaccttcg caaaaaatat ttattatgaa aatacatttt ttaacctatg
     6361 tacaactgat ggaaaagcag gaaccaatga aaacttaata aatagatatg cattaggatt
     6421 tgattttgac aaaaaagaat taggacaagg ttttaattat aaagatataa ttaatttatt
     6481 tactaagata ggattacatt atcatatcct agttgatagt ggaaatggat tccatgttta
     6541 tgtgctaatt aataaaacta ataacattaa gttagtatca gaagttacaa atacattaat
     6601 aaataaattg ggtgcagata aacaagcaaa tttatctact caagtattaa gagtacctta
     6661 tacatataat attaaaaata ctactaaaca agtaaaaata atacaccaag acaaaaatat
     6721 atatagatat gacatagaaa agttagctaa aaaatattgc aaagatgtaa aaacagtagg
     6781 taatactaat acaaaatata tattagatag taagctacca aattgtatag tagatatttt
     6841 aaaaaatggt agtaaagatg gacataaaaa cctagatttg caaaaaatag ttgtgacttt
     6901 aagattgagg aataaaagtt taagtcaagt aatatccgtt gctagagaat ggaactatat
     6961 atcacaaaat agtctttcaa atagtgagct agaatatcaa gtcaagtata tgtatgagaa
     7021 gcttaaaacg gttaattttg gttgtacagg ttgtgagttt aatagtgatt gttggaataa
     7081 aatagaatca gattttatat atagtgatga agatactttg ttcaatatgc cacataagca
     7141 ctcaaaggat ttgaaatata agaataggaa aggggttaaa ataatgactg gtaatcaatt
     7201 gtttatctat aatgtgttac ttaacaataa agatagagaa ttaaacatag acgatataat
     7261 ggagctgata acctataaac gtaagaagaa agttaaaaac attgttatga gtgaaaagac
     7321 attaagagaa acattaaaag aacttcaaca taatgattat attacaaaaa caaaaggtgt
     7381 tacaaagcta ggaataaaag atacatacaa tgtaaaagaa gttagatgta atatagataa
     7441 acaatatact attagttact ttgttaccat ggcagtaatt tggggaataa tttcaactga
     7501 agaattaaga ttatatactc atatgagata taagcaagat ttattggtca aagatgataa
     7561 aataaaagga aatatattaa gaattaatca agaggaatta gcaaaagatt taggagtaac
     7621 acagcaaaga atttcaaata tgatagaatc tttattagat actaaaattt tagatgtatg
     7681 ggaaactaaa ataaatgata gaggatttat gtactataca tatagattaa acaagtagat
     7741 ttttgatagg attagaattg attttctagt cctattttta tgcaaaaaaa ctaattaata
     7801 aaaatttctt ttggtaaaat aattgtacga gaattgcaaa aaaaaaatgg catcaaagta
     7861 ttgaaattaa gccgttttaa aaatt
//
