LOCUS Exported 7911 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION E. coli-C. difficile shuttle vector: tetracycline-inducible promoter (Ptet) driving expression of red fluorescent protein (mCherryOpt). ACCESSION . VERSION . KEYWORDS pDSW1728 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7911 /organism="synthetic DNA construct" /mol_type="other DNA" 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..7868) /codon_start=1 /gene="tetR from transposon Tn10" /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="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS" 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 //