LOCUS       sequence-235839-        7911 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli-C. difficile shuttle vector: tetracycline-inducible promoter
            (Ptet) driving expression of red fluorescent protein (mCherryOpt).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7911)
  AUTHORS   Ransom EM, Ellermeier CD, Weiss DS
  TITLE     Use of mCherry Red fluorescent protein for studies of protein 
            localization and gene expression in Clostridium difficile.
  JOURNAL   Appl Environ Microbiol. 2015 Mar;81(5):1652-60. doi: 
            10.1128/AEM.03446-14. Epub 2014 Dec 19.
  PUBMED    25527559
REFERENCE   2  (bases 1 to 7911)
  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/AEM.03446-14"; journalName: "Appl Environ Microbiol"; date:
            "2015-03"; volume: "81"; issue: "5"; pages: "1652-60"
COMMENT     Sequence Label: pDSW1728
FEATURES             Location/Qualifiers
     source          1..7911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    67..85
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     CDS             complement(1206..1577)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            1610..1719
                     /note="incP origin of transfer"
     primer_bind     complement(2910..2927)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3081..3669)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3161..3180)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7245..7871)
                     /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     7776..7795
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
ORIGIN
        1 tatttgtcaa actagttttt tatttcgatg ccctggactt catgaaaaac taaaaaaaat
       61 attgacactc tatcattgat agagtataat taaaataagc ttgatcgtag cgttaacaga
      121 tctgagctcc tgcagtaaag gagaaaattt tatggtatct aaaggagaag aagataatat
      181 ggctataatt aaagaattta tgagatttaa agttcatatg gaaggatctg ttaatggaca
      241 tgaatttgaa atagaaggag aaggagaagg aagaccttat gaaggtactc aaacagctaa
      301 attaaaagta actaaaggtg gtccattacc atttgcatgg gatatattaa gtcctcaatt
      361 tatgtatgga tctaaagcat atgttaaaca tcctgctgat atacctgatt atttaaaatt
      421 atcatttcca gaaggtttta aatgggaaag agttatgaat tttgaagatg gtggagtagt
      481 tactgtaaca caagatagtt cattacaaga tggagatttt atatataaag ttaaattaag
      541 aggtactaat tttccatctg atggaccagt aatgcaaaag aaaactatgg gatgggaagc
      601 atcatctgaa agaatgtatc cagaagatgg agcattaaaa ggtgaaatta aacaaagatt
      661 aaaattaaaa gatggaggtc attatgatgc tgaagttaag actacatata aagctaagaa
      721 accagttcaa ttaccaggtg catataatgt taatattaaa ttagatataa cttctcataa
      781 tgaagattat actatagttg aacaatatga aagagcagaa ggaagacatt caacaggagg
      841 tatggatgaa ttatataaat aaggatccta taagttttaa taaaacttta aatagaaaaa
      901 ggcttctctc atgagaagtc ttttttattt aaaataaata taaaataaaa tagaggctat
      961 aaatagcctc tattttatgt gagaaatccc taaataaaaa gatgccagtg tgctggaatt
     1021 cgcccttagg ctgcgctagg gacctcttta gctccttgga agctgtcagt agtaggggat
     1081 cggtcttgcc ttgctcgtcg gtgatgtact tcaccagctc cgcgaagtcg ctcttcttga
     1141 tggagcgcat ggggacgtgc ttggcaatca cgcgcacccc ccggccgttt tagcggctaa
     1201 aaaagtcatg gctctgccct cgggcggacc acgcccatca tgaccttgcc aagctcgtcc
     1261 tgcttctctt cgatcttcgc cagcagggcg aggatcgtgg catcaccgaa ccgcgccgtg
     1321 cgcgggtcgt cggtgagcca gagtttcagc aggccgccca ggcggcccag gtcgccattg
     1381 atgcgggcca gctcgcggac gtgctcatag tccacgacgc ccgtgatttt gtagccctgg
     1441 ccgacggcca gcaggtaggc cgacaggctc atgccggccg ccgccgcctt ttcctcaatc
     1501 gctcttcgtt cgtctggaag gcagtacacc ttgataggtg ggctgccctt cctggttggc
     1561 ttggtttcat cagccatccg cttgccctca tctgttacgc cggcggtagc cggccagcct
     1621 cgcagagcag gattcccgtt gagcaccgcc aggtgcgaat aagggacagt gaagaaggaa
     1681 cacccgctcg cgggtgggcc tacttcacct atcctgcccg gctgacgccg ttggatacac
     1741 caaggaaagt ctacacgaac cctttggcaa aatcctgtat atcgtgcgaa aaaggatgga
     1801 tataccgaaa aaatcgctat aatgaccccg aagcaggaag ggcgaattct gcagatctga
     1861 ccggtctctg aaaatataaa aaccacagat tgatactaaa accttggttg tgttgctttt
     1921 cggggcttaa atcaaggaaa aatccttgtt ttaagccttt caaaaagaaa cacaaggtct
     1981 ttgtactaac ctgtggttat gtataaaatt gtagatttta gggtaacaaa aaacaccgta
     2041 tttctacgat gtttttgctt aaatacttgt ttttagttac agacaaacct gaagttaact
     2101 atttatcaat tcctgcaatt cgtttacaaa acggcaaatg tgaaatccgt cacatactgc
     2161 gtgatgaact tgaattgcca aaggaagtat aattttgtta tcttctttat aatatttccc
     2221 catagtaaaa ataggaatca aataatcata tcctttctgc aaattcagat taaagccatc
     2281 gaaggttgac cacggtatca tagatacatt aaaaatgttt tccggagcat ttggctttcc
     2341 ttccattcta tgattgtttc cataccgttg cgtatcactt tcataatctg ctaaaaatga
     2401 tttaaagtca gacttacact cagtccaaag gctggaaaat gtttcagtat cattgtgaaa
     2461 tattgtatag cttggtatca tctcatcata tatccccaat tcaccatctt gattgattgc
     2521 cgtcctaaac tctgaatggc ggtttacaat cattgcaata taataaagca ttgcaggata
     2581 tagtttcatt cccttttcct ttatttgtgt gatatccact ttaacggtca tgctgtaggt
     2641 acaaggtaca cttgcaaagt agtggtcaaa atactctttt ctgttccaac tatttttatc
     2701 aattttttca aataccatct aagttccctc tcaaattcaa gtttatcgct ctaatgaaca
     2761 aagatattat accacatttt tgtgaatttt tcaacttgcc cacttcgact gcactcccga
     2821 cttaataact tcttgaacac ttgccgaaaa agaaaaactg ccgggtacgt acccggggat
     2881 cgatccccgg ccgagcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2941 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3001 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3061 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3121 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3181 ccttgaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3241 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3301 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3361 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3421 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3481 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3541 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3601 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3661 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3721 tcacgttaag ggattttggt catgaaatgc aagtttctaa ctaacataca tcatagttac
     3781 taaactatgg taactataat tttattaact atattaagca ctgattagta ctataactca
     3841 atataagcat atcccctgta tcgtaactag agaaccaaac gacggaaaaa gcgatataga
     3901 taagtttaga tagtgcacca attttttact tttctatttt agaatctagc atttccaatg
     3961 cttactaata ctatttttga tttttaactt gttttaaaaa agcacctact aattaggatt
     4021 ctagctctat attctagtta tacattctct atattctagc tctatattct agttatacat
     4081 tctctatatt ctagctctat attctagtta tacattctct atattctagc tctatattct
     4141 agttatacat tctctatatt ctagctctat attctagtta tacattctct atattctagc
     4201 tctatattct agttatacat tctctatatt ctagctctat attctagtta tacattctct
     4261 aaaaaacata tatatatata ctacatatat atgtacaatt attttaccaa aagaaatttt
     4321 taaaacggct taatttcaat actttgatgc catttttttt ttgcaattct cgtacaatta
     4381 ttttaccaaa agaaattttt attaattagt ttttttgcat aaaaatagga ctagaaaatc
     4441 aattctaatc ctatcaaaaa tctacttgtt taatctatat gtatagtaca taaatcctct
     4501 atcatttatt ttagtttccc atacatctaa aattttagta tctaataaag attctatcat
     4561 atttgaaatt ctttgctgtg ttactcctaa atcttttgct aattcctctt gattaattct
     4621 taatatattt ccttttattt tatcatcttt gaccaataaa tcttgcttat atctcatatg
     4681 agtatataat cttaattctt cagttgaaat tattccccaa attactgcca tggtaacaaa
     4741 gtaactaata gtatattgtt tatctatatt acatctaact tcttttacat tgtatgtatc
     4801 ttttattcct agctttgtaa caccttttgt ttttgtaata taatcattat gttgaagttc
     4861 ttttaatgtt tctcttaatg tcttttcact cataacaatg tttttaactt tcttcttacg
     4921 tttataggtt atcagctcca ttatatcgtc tatgtttaat tctctatctt tattgttaag
     4981 taacacatta tagataaaca attgattacc agtcattatt ttaacccctt tcctattctt
     5041 atatttcaaa tcctttgagt gcttatgtgg catattgaac aaagtatctt catcactata
     5101 tataaaatct gattctattt tattccaaca atcactatta aactcacaac ctgtacaacc
     5161 aaaattaacc gttttaagct tctcatacat atacttgact tgatattcta gctcactatt
     5221 tgaaagacta ttttgtgata tatagttcca ttctctagca acggatatta cttgacttaa
     5281 acttttattc ctcaatctta aagtcacaac tattttttgc aaatctaggt ttttatgtcc
     5341 atctttacta ccatttttta aaatatctac tatacaattt ggtagcttac tatctaatat
     5401 atattttgta ttagtattac ctactgtttt tacatctttg caatattttt tagctaactt
     5461 ttctatgtca tatctatata tatttttgtc ttggtgtatt atttttactt gtttagtagt
     5521 atttttaata ttatatgtat aaggtactct taatacttga gtagataaat ttgcttgttt
     5581 atctgcaccc aatttattta ttaatgtatt tgtaacttct gatactaact taatgttatt
     5641 agttttatta attagcacat aaacatggaa tccatttcca ctatcaacta ggatatgata
     5701 atgtaatcct atcttagtaa ataaattaat tatatcttta taattaaaac cttgtcctaa
     5761 ttcttttttg tcaaaatcaa atcctaatgc atatctattt attaagtttt cattggttcc
     5821 tgcttttcca tcagttgtac ataggttaaa aaatgtattt tcataataaa tattttttgc
     5881 gaaggtaaat tctactatat catctatatt tttaaaaaaa gttgttttac tatattcttt
     5941 aaattcttct ccgagttcta attgtttact attactttta ttattttgaa atactctaac
     6001 atattcatta tcaataagat tatttccttg tcctatacca tctctaaata ctgccattag
     6061 aaattttttc aacttatatt ttgaatctaa ttgttccata tatttattat ctccttatct
     6121 agttttttta agagataaca ctacattaaa tatatactta ttgactaatc aacttttaca
     6181 actattgaat aattttagct tagtatgtat aataatattg ttaagatgct aaaattgata
     6241 gcaatattat tatataacta cagatactta atcagttaga ccgatattgg agttggtagc
     6301 tccataaact caattggtag ttgagttgtc attgtctagt agtattatct cattttgttg
     6361 ttatcttaaa atattatttt aaaaatactc tgcttggtcg gcttagggta ttttttttgt
     6421 ttaattccaa tattggttca aactcctgtg taatactttt gacacttcca tctgtattga
     6481 attgacagtt gatatggtat ctcattattg gcttgttctt atgtttttct atgtatttat
     6541 tttttacttt caaataattt gtatttccat ttttagttat gtttaacaca ctatccttat
     6601 tactcaataa aattattttt tgttttaaca tatgatttat agaatctaaa aattgatatt
     6661 gacttataat agtactttct acctctctag ctttttctag tgtttcaaat tcttcaaact
     6721 ctgctatatt gctatagtat agcttatcat taataatatt atctgacaac tttaattcca
     6781 ttttataaaa caattccaag ttatgtataa aatttttaat gttacaacat atagacttat
     6841 tatttgttct tatatggtat ctccatttac tacttctagc tctcatgtta tatgcacaac
     6901 tactcccaac ataaacatat attgcttgtt ctaactcata tttttgtaaa atttcaacat
     6961 catatccagt caaatcgtac tctctcttat atacaaatgg tttttctaat tttttgttca
     7021 ataaatctcc tcctatttct tacttaattt agctaaaaat ctatttatat ttcacctcct
     7081 ttttgacttt aagcctacga ataccatatt tatgattata acatcttcta tttttcccaa
     7141 atccttacat ctcccctaat ttttagactt aagggcgaat tctgcatcaa gctagcataa
     7201 aaataagaag cctgcatttg caggcttctt atttttatgg taccttaaga cccactttca
     7261 catttaagtt gtttttctaa tccgcatatg atcaattcaa ggccgaataa gaaggctggc
     7321 tctgcacctt ggtgatcaaa taattcgata gcttgtcgta ataatggcgg catactatca
     7381 gtagtaggtg tttccctttc ttctttagcg acttgatgct cttgatcttc caatacgcaa
     7441 cctaaagtaa aatgccccac agcgctgagt gcatataatg cattctctag tgaaaaacct
     7501 tgttggcata aaaaggctaa ttgattttcg agagtttcat actgtttttc tgtaggccgt
     7561 gtacctaaat gtacttttgc tccatcgcga tgacttagta aagcacatct aaaactttta
     7621 gcgttattac gtaaaaaatc ttgccagctt tccccttcta aagggcaaaa gtgagtatgg
     7681 tgcctatcta acatctcaat ggctaaggcg tcgagcaaag cccgcttatt ttttacatgc
     7741 caatacaatg taggctgctc tacacctagc ttctgggcga gtttacgggt tgttaaacct
     7801 tcgattccga cctcattaag cagctctaat gcgctgttaa tcactttact tttatctaat
     7861 ctagacatca ttaattcctc ctttttgttg acattatatc attgatagag t
//
