LOCUS       V007836                 7913 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V007836
VERSION     V007836
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7913)
  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. 81 (5), 1652-1660 (2015)
   PUBMED   25527559
REFERENCE   2  (bases 1 to 7913)
  AUTHORS   Ransom EM, Ellermeier CD, Weiss DS.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-AUG-2015) Microbiology, The University of Iowa, 51
            Newton Rd, Iowa City, IA 52242, USA
REFERENCE   3  (bases 1 to 7913)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7913)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
            Environ. Microbiol."; date: "2015"; volume: "81"; issue: "5"; pages:
            "1652-1660"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (06-AUG-2015) Microbiology, The University of Iowa, 51 Newton Rd,
            Iowa City, IA 52242, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7913
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(58..681)
                     /label="TetR"
                     /note="tetracycline repressor TetR"
     protein_bind    788..806
                     /label="tet operator"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     CDS             873..1580
                     /label="mCherry"
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     CDS             complement(1932..2300)
                     /label="traJ"
                     /note="oriT-recognizing protein"
     oriT            complement(2333..2442)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             complement(2821..3441)
                     /gene="catP"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from Clostridium
                     perfringens. Accession#: P26826"
     rep_origin      complement(3804..4392)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5185..6822)
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label="repA"
                     /note="plasmid replication; derived from Clostridium
                     difficile"
                     /protein_id="AMA07799.1"
                     /translation="MEQLDSKYKLKKFLMAVFRDGIGQGNNLIDNEYVRVFQNNKSNSK
                     QLELGEEFKEYSKTTFFKNIDDIVEFTFAKNIYYENTFFNLCTTDGKAGTNENLINRYA
                     LGFDFDKKELGQGFNYKDIINLFTKIGLHYHILVDSGNGFHVYVLINKTNNIKLVSEVT
                     NTLINKLGADKQANLSTQVLRVPYTYNIKNTTKQVKIIHQDKNIYRYDIEKLAKKYCKD
                     VKTVGNTNTKYILDSKLPNCIVDILKNGSKDGHKNLDLQKIVVTLRLRNKSLSQVISVA
                     REWNYISQNSLSNSELEYQVKYMYEKLKTVNFGCTGCEFNSDCWNKIESDFIYSDEDTL
                     FNMPHKHSKDLKYKNRKGVKIMTGNQLFIYNVLLNNKDRELNIDDIMELITYKRKKKVK
                     NIVMSEKTLRETLKELQHNDYITKTKGVTKLGIKDTYNVKEVRCNIDKQYTISYFVTMA
                     VIWGIISTEELRLYTHMRYKQDLLVKDDKIKGNILRINQEELAKDLGVTQQRISNMIES
                     LLDTKILDVWETKINDRGFMYYTYRLNK"
     gene            complement(5185..6822)
                     /gene="repA"
                     /label="repA"
     CDS             complement(7097..7609)
                     /codon_start=1
                     /gene="orfB"
                     /product="OrfB"
                     /label="orfB"
                     /protein_id="AMA07800.1"
                     /translation="MRARSSKWRYHIRTNNKSICCNIKNFIHNLELFYKMELKLSDNII
                     NDKLYYSNIAEFEEFETLEKAREVESTIISQYQFLDSINHMLKQKIILLSNKDSVLNIT
                     KNGNTNYLKVKNKYIEKHKNKPIMRYHINCQFNTDGSVKSITQEFEPILELNKKNTLSR
                     PSRVFLK"
     gene            complement(7097..7609)
                     /gene="orfB"
                     /label="orfB"
ORIGIN
        1 gctagcataa aaataagaag cctgcatttg caggcttctt atttttatgg taccttaaga
       61 cccactttca catttaagtt gtttttctaa tccgcatatg atcaattcaa ggccgaataa
      121 gaaggctggc tctgcacctt ggtgatcaaa taattcgata gcttgtcgta ataatggcgg
      181 catactatca gtagtaggtg tttccctttc ttctttagcg acttgatgct cttgatcttc
      241 caatacgcaa cctaaagtaa aatgccccac agcgctgagt gcatataatg cattctctag
      301 tgaaaaacct tgttggcata aaaaggctaa ttgattttcg agagtttcat actgtttttc
      361 tgtaggccgt gtacctaaat gtacttttgc tccatcgcga tgacttagta aagcacatct
      421 aaaactttta gcgttattac gtaaaaaatc ttgccagctt tccccttcta aagggcaaaa
      481 gtgagtatgg tgcctatcta acatctcaat ggctaaggcg tcgagcaaag cccgcttatt
      541 ttttacatgc caatacaatg taggctgctc tacacctagc ttctgggcga gtttacgggt
      601 tgttaaacct tcgattccga cctcattaag cagctctaat gcgctgttaa tcactttact
      661 tttatctaat ctagacatca ttaattcctc ctttttgttg acattatatc attgatagag
      721 ttatttgtca aactagtttt ttatttcgat gccctggact tcatgaaaaa ctaaaaaaaa
      781 tattgacact ctatcattga tagagtataa ttaaaataag cttgatcgta gcgttaacag
      841 atctgagctc ctgcagtaaa ggagaaaatt ttatggtatc taaaggagaa gaagataata
      901 tggctataat taaagaattt atgagattta aagttcatat ggaaggatct gttaatggac
      961 atgaatttga aatagaagga gaaggagaag gaagacctta tgaaggtact caaacagcta
     1021 aattaaaagt aactaaaggt ggtccattac catttgcatg ggatatatta agtcctcaat
     1081 ttatgtatgg atctaaagca tatgttaaac atcctgctga tatacctgat tatttaaaat
     1141 tatcatttcc agaaggtttt aaatgggaaa gagttatgaa ttttgaagat ggtggagtag
     1201 ttactgtaac acaagatagt tcattacaag atggagattt tatatataaa gttaaattaa
     1261 gaggtactaa ttttccatct gatggaccag taatgcaaaa gaaaactatg ggatgggaag
     1321 catcatctga aagaatgtat ccagaagatg gagcattaaa aggtgaaatt aaacaaagat
     1381 taaaattaaa agatggaggt cattatgatg ctgaagttaa gactacatat aaagctaaga
     1441 aaccagttca attaccaggt gcatataatg ttaatattaa attagatata acttctcata
     1501 atgaagatta tactatagtt gaacaatatg aaagagcaga aggaagacat tcaacaggag
     1561 gtatggatga attatataaa taaggatcct ataagtttta ataaaacttt aaatagaaaa
     1621 aggcttctct catgagaagt cttttttatt taaaataaat ataaaataaa atagaggcta
     1681 taaatagcct ctattttatg tgagaaatcc ctaaataaaa agatgccagt gtgtgctgga
     1741 attcgccctt aggctgcgct agggacctct ttagctcctt ggaagctgtc agtagtaggg
     1801 gatcggtctt gccttgctcg tcggtgatgt acttcaccag ctccgcgaag tcgctcttct
     1861 tgatggagcg catggggacg tgcttggcaa tcacgcgcac cccccggccg ttttagcggc
     1921 taaaaaagtc atggctctgc cctcgggcgg accacgccca tcatgacctt gccaagctcg
     1981 tcctgcttct cttcgatctt cgccagcagg gcgaggatcg tggcatcacc gaaccgcgcc
     2041 gtgcgcgggt cgtcggtgag ccagagtttc agcaggccgc ccaggcggcc caggtcgcca
     2101 ttgatgcggg ccagctcgcg gacgtgctca tagtccacga cgcccgtgat tttgtagccc
     2161 tggccgacgg ccagcaggta ggccgacagg ctcatgccgg ccgccgccgc cttttcctca
     2221 atcgctcttc gttcgtctgg aaggcagtac accttgatag gtgggctgcc cttcctggtt
     2281 ggcttggttt catcagccat ccgcttgccc tcatctgtta cgccggcggt agccggccag
     2341 cctcgcagag caggattccc gttgagcacc gccaggtgcg aataagggac agtgaagaag
     2401 gaacacccgc tcgcgggtgg gcctacttca cctatcctgc ccggctgacg ccgttggata
     2461 caccaaggaa agtctacacg aaccctttgg caaaatcctg tatatcgtgc gaaaaaggat
     2521 ggatataccg aaaaaatcgc tataatgacc ccgaagcagg aagggcgaat tctgcagatc
     2581 tgaccggtct ctgaaaatat aaaaaccaca gattgatact aaaaccttgg ttgtgttgct
     2641 tttcggggct taaatcaagg aaaaatcctt gttttaagcc tttcaaaaag aaacacaagg
     2701 tctttgtact aacctgtggt tatgtataaa attgtagatt ttagggtaac aaaaaacacc
     2761 gtatttctac gatgtttttg cttaaatact tgtttttagt tacagacaaa cctgaagtta
     2821 actatttatc aattcctgca attcgtttac aaaacggcaa atgtgaaatc cgtcacatac
     2881 tgcgtgatga acttgaattg ccaaaggaag tataattttg ttatcttctt tataatattt
     2941 ccccatagta aaaataggaa tcaaataatc atatcctttc tgcaaattca gattaaagcc
     3001 atcgaaggtt gaccacggta tcatagatac attaaaaatg ttttccggag catttggctt
     3061 tccttccatt ctatgattgt ttccataccg ttgcgtatca ctttcataat ctgctaaaaa
     3121 tgatttaaag tcagacttac actcagtcca aaggctggaa aatgtttcag tatcattgtg
     3181 aaatattgta tagcttggta tcatctcatc atatatcccc aattcaccat cttgattgat
     3241 tgccgtccta aactctgaat ggcggtttac aatcattgca atataataaa gcattgcagg
     3301 atatagtttc attccctttt cctttatttg tgtgatatcc actttaacgg tcatgctgta
     3361 ggtacaaggt acacttgcaa agtagtggtc aaaatactct tttctgttcc aactattttt
     3421 atcaattttt tcaaatacca tctaagttcc ctctcaaatt caagtttatc gctctaatga
     3481 acaaagatat tataccacat ttttgtgaat ttttcaactt gcccacttcg actgcactcc
     3541 cgacttaata acttcttgaa cacttgccga aaaagaaaaa ctgccgggta cgtacccggg
     3601 gatcgatccc cggccgagcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3661 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3721 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3781 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3841 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3901 cccccttgaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3961 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4021 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4081 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4141 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4201 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4261 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4321 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4381 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4441 aactcacgtt aagggatttt ggtcatgaaa tgcaagtttc taactaacat acatcatagt
     4501 tactaaacta tggtaactat aattttatta actatattaa gcactgatta gtactataac
     4561 tcaatataag catatcccct gtatcgtaac tagagaacca aacgacggaa aaagcgatat
     4621 agataagttt agatagtgca ccaatttttt acttttctat tttagaatct agcatttcca
     4681 atgcttacta atactatttt tgatttttaa cttgttttaa aaaagcacct actaattagg
     4741 attctagctc tatattctag ttatacattc tctatattct agctctatat tctagttata
     4801 cattctctat attctagctc tatattctag ttatacattc tctatattct agctctatat
     4861 tctagttata cattctctat attctagctc tatattctag ttatacattc tctatattct
     4921 agctctatat tctagttata cattctctat attctagctc tatattctag ttatacattc
     4981 tctaaaaaac atatatatat atactacata tatatgtaca attattttac caaaagaaat
     5041 ttttaaaacg gcttaatttc aatactttga tgccattttt tttttgcaat tctcgtacaa
     5101 ttattttacc aaaagaaatt tttattaatt agtttttttg cataaaaata ggactagaaa
     5161 atcaattcta atcctatcaa aaatctactt gtttaatcta tatgtatagt acataaatcc
     5221 tctatcattt attttagttt cccatacatc taaaatttta gtatctaata aagattctat
     5281 catatttgaa attctttgct gtgttactcc taaatctttt gctaattcct cttgattaat
     5341 tcttaatata tttcctttta ttttatcatc tttgaccaat aaatcttgct tatatctcat
     5401 atgagtatat aatcttaatt cttcagttga aattattccc caaattactg ccatggtaac
     5461 aaagtaacta atagtatatt gtttatctat attacatcta acttctttta cattgtatgt
     5521 atcttttatt cctagctttg taacaccttt tgtttttgta atataatcat tatgttgaag
     5581 ttcttttaat gtttctctta atgtcttttc actcataaca atgtttttaa ctttcttctt
     5641 acgtttatag gttatcagct ccattatatc gtctatgttt aattctctat ctttattgtt
     5701 aagtaacaca ttatagataa acaattgatt accagtcatt attttaaccc ctttcctatt
     5761 cttatatttc aaatcctttg agtgcttatg tggcatattg aacaaagtat cttcatcact
     5821 atatataaaa tctgattcta ttttattcca acaatcacta ttaaactcac aacctgtaca
     5881 accaaaatta accgttttaa gcttctcata catatacttg acttgatatt ctagctcact
     5941 atttgaaaga ctattttgtg atatatagtt ccattctcta gcaacggata ttacttgact
     6001 taaactttta ttcctcaatc ttaaagtcac aactattttt tgcaaatcta ggtttttatg
     6061 tccatcttta ctaccatttt ttaaaatatc tactatacaa tttggtagct tactatctaa
     6121 tatatatttt gtattagtat tacctactgt ttttacatct ttgcaatatt ttttagctaa
     6181 cttttctatg tcatatctat atatattttt gtcttggtgt attattttta cttgtttagt
     6241 agtattttta atattatatg tataaggtac tcttaatact tgagtagata aatttgcttg
     6301 tttatctgca cccaatttat ttattaatgt atttgtaact tctgatacta acttaatgtt
     6361 attagtttta ttaattagca cataaacatg gaatccattt ccactatcaa ctaggatatg
     6421 ataatgtaat cctatcttag taaataaatt aattatatct ttataattaa aaccttgtcc
     6481 taattctttt ttgtcaaaat caaatcctaa tgcatatcta tttattaagt tttcattggt
     6541 tcctgctttt ccatcagttg tacataggtt aaaaaatgta ttttcataat aaatattttt
     6601 tgcgaaggta aattctacta tatcatctat atttttaaaa aaagttgttt tactatattc
     6661 tttaaattct tctccgagtt ctaattgttt actattactt ttattatttt gaaatactct
     6721 aacatattca ttatcaataa gattatttcc ttgtcctata ccatctctaa atactgccat
     6781 tagaaatttt ttcaacttat attttgaatc taattgttcc atatatttat tatctcctta
     6841 tctagttttt ttaagagata acactacatt aaatatatac ttattgacta atcaactttt
     6901 acaactattg aataatttta gcttagtatg tataataata ttgttaagat gctaaaattg
     6961 atagcaatat tattatataa ctacagatac ttaatcagtt agaccgatat tggagttggt
     7021 agctccataa actcaattgg tagttgagtt gtcattgtct agtagtatta tctcattttg
     7081 ttgttatctt aaaatattat tttaaaaata ctctgcttgg tcggcttagg gtattttttt
     7141 tgtttaattc caatattggt tcaaactcct gtgtaatact tttgacactt ccatctgtat
     7201 tgaattgaca gttgatatgg tatctcatta ttggcttgtt cttatgtttt tctatgtatt
     7261 tattttttac tttcaaataa tttgtatttc catttttagt tatgtttaac acactatcct
     7321 tattactcaa taaaattatt ttttgtttta acatatgatt tatagaatct aaaaattgat
     7381 attgacttat aatagtactt tctacctctc tagctttttc tagtgtttca aattcttcaa
     7441 actctgctat attgctatag tatagcttat cattaataat attatctgac aactttaatt
     7501 ccattttata aaacaattcc aagttatgta taaaattttt aatgttacaa catatagact
     7561 tattatttgt tcttatatgg tatctccatt tactacttct agctctcatg ttatatgcac
     7621 aactactccc aacataaaca tatattgctt gttctaactc atatttttgt aaaatttcaa
     7681 catcatatcc agtcaaatcg tactctctct tatatacaaa tggtttttct aattttttgt
     7741 tcaataaatc tcctcctatt tcttacttaa tttagctaaa aatctattta tatttcacct
     7801 cctttttgac tttaagccta cgaataccat atttatgatt ataacatctt ctatttttcc
     7861 caaatcctta catctcccct aatttttaga cttaagggcg aattctgcat caa
//
