LOCUS       V003825                 7952 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003825
VERSION     V003825
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7952)
  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 7952)
  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 7952)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7952)
  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..7952
                     /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(1971..2339)
                     /label="traJ"
                     /note="oriT-recognizing protein"
     oriT            complement(2372..2481)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             complement(2860..3480)
                     /gene="catP"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from Clostridium
                     perfringens. Accession#: P26826"
     rep_origin      complement(3843..4431)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5224..6861)
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label="repA"
                     /note="plasmid replication; derived from Clostridium
                     difficile"
                     /protein_id="AMA07802.1"
                     /translation="MEQLDSKYKLKKFLMAVFRDGIGQGNNLIDNEYVRVFQNNKSNSK
                     QLELGEEFKEYSKTTFFKNIDDIVEFTFAKNIYYENTFFNLCTTDGKAGTNENLINRYA
                     LGFDFDKKELGQGFNYKDIINLFTKIGLHYHILVDSGNGFHVYVLINKTNNIKLVSEVT
                     NTLINKLGADKQANLSTQVLRVPYTYNIKNTTKQVKIIHQDKNIYRYDIEKLAKKYCKD
                     VKTVGNTNTKYILDSKLPNCIVDILKNGSKDGHKNLDLQKIVVTLRLRNKSLSQVISVA
                     REWNYISQNSLSNSELEYQVKYMYEKLKTVNFGCTGCEFNSDCWNKIESDFIYSDEDTL
                     FNMPHKHSKDLKYKNRKGVKIMTGNQLFIYNVLLNNKDRELNIDDIMELITYKRKKKVK
                     NIVMSEKTLRETLKELQHNDYITKTKGVTKLGIKDTYNVKEVRCNIDKQYTISYFVTMA
                     VIWGIISTEELRLYTHMRYKQDLLVKDDKIKGNILRINQEELAKDLGVTQQRISNMIES
                     LLDTKILDVWETKINDRGFMYYTYRLNK"
     gene            complement(5224..6861)
                     /gene="repA"
                     /label="repA"
     CDS             complement(7136..7648)
                     /codon_start=1
                     /gene="OrfB"
                     /product="orfB"
                     /label="OrfB"
                     /protein_id="AMA07801.1"
                     /translation="MRARSSKWRYHIRTNNKSICCNIKNFIHNLELFYKMELKLSDNII
                     NDKLYYSNIAEFEEFETLEKAREVESTIISQYQFLDSINHMLKQKIILLSNKDSVLNIT
                     KNGNTNYLKVKNKYIEKHKNKPIMRYHINCQFNTDGSVKSITQEFEPILELNKKNTLSR
                     PSRVFLK"
     gene            complement(7136..7648)
                     /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 atgaacgcat gcgtcgactt aattaaacag ctgaggcgcg
     1621 ccggatccta taagttttaa taaaacttta aatagaaaaa ggcttctctc atgagaagtc
     1681 ttttttattt aaaataaata taaaataaaa tagaggctat aaatagcctc tattttatgt
     1741 gagaaatccc taaataaaaa gatgccagtg tgtgctggaa ttcgccctta ggctgcgcta
     1801 gggacctctt tagctccttg gaagctgtca gtagtagggg atcggtcttg ccttgctcgt
     1861 cggtgatgta cttcaccagc tccgcgaagt cgctcttctt gatggagcgc atggggacgt
     1921 gcttggcaat cacgcgcacc ccccggccgt tttagcggct aaaaaagtca tggctctgcc
     1981 ctcgggcgga ccacgcccat catgaccttg ccaagctcgt cctgcttctc ttcgatcttc
     2041 gccagcaggg cgaggatcgt ggcatcaccg aaccgcgccg tgcgcgggtc gtcggtgagc
     2101 cagagtttca gcaggccgcc caggcggccc aggtcgccat tgatgcgggc cagctcgcgg
     2161 acgtgctcat agtccacgac gcccgtgatt ttgtagccct ggccgacggc cagcaggtag
     2221 gccgacaggc tcatgccggc cgccgccgcc ttttcctcaa tcgctcttcg ttcgtctgga
     2281 aggcagtaca ccttgatagg tgggctgccc ttcctggttg gcttggtttc atcagccatc
     2341 cgcttgccct catctgttac gccggcggta gccggccagc ctcgcagagc aggattcccg
     2401 ttgagcaccg ccaggtgcga ataagggaca gtgaagaagg aacacccgct cgcgggtggg
     2461 cctacttcac ctatcctgcc cggctgacgc cgttggatac accaaggaaa gtctacacga
     2521 accctttggc aaaatcctgt atatcgtgcg aaaaaggatg gatataccga aaaaatcgct
     2581 ataatgaccc cgaagcagga agggcgaatt ctgcagatct gaccggtctc tgaaaatata
     2641 aaaaccacag attgatacta aaaccttggt tgtgttgctt ttcggggctt aaatcaagga
     2701 aaaatccttg ttttaagcct ttcaaaaaga aacacaaggt ctttgtacta acctgtggtt
     2761 atgtataaaa ttgtagattt tagggtaaca aaaaacaccg tatttctacg atgtttttgc
     2821 ttaaatactt gtttttagtt acagacaaac ctgaagttaa ctatttatca attcctgcaa
     2881 ttcgtttaca aaacggcaaa tgtgaaatcc gtcacatact gcgtgatgaa cttgaattgc
     2941 caaaggaagt ataattttgt tatcttcttt ataatatttc cccatagtaa aaataggaat
     3001 caaataatca tatcctttct gcaaattcag attaaagcca tcgaaggttg accacggtat
     3061 catagataca ttaaaaatgt tttccggagc atttggcttt ccttccattc tatgattgtt
     3121 tccataccgt tgcgtatcac tttcataatc tgctaaaaat gatttaaagt cagacttaca
     3181 ctcagtccaa aggctggaaa atgtttcagt atcattgtga aatattgtat agcttggtat
     3241 catctcatca tatatcccca attcaccatc ttgattgatt gccgtcctaa actctgaatg
     3301 gcggtttaca atcattgcaa tataataaag cattgcagga tatagtttca ttcccttttc
     3361 ctttatttgt gtgatatcca ctttaacggt catgctgtag gtacaaggta cacttgcaaa
     3421 gtagtggtca aaatactctt ttctgttcca actattttta tcaatttttt caaataccat
     3481 ctaagttccc tctcaaattc aagtttatcg ctctaatgaa caaagatatt ataccacatt
     3541 tttgtgaatt tttcaacttg cccacttcga ctgcactccc gacttaataa cttcttgaac
     3601 acttgccgaa aaagaaaaac tgccgggtac gtacccgggg atcgatcccc ggccgagcgc
     3661 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3721 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3781 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3841 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3901 tggcgaaacc cgacaggact ataaagatac caggcgtttc ccccttgaag ctccctcgtg
     3961 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4021 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4081 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4141 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4201 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4261 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4321 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4381 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4441 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4501 gtcatgaaat gcaagtttct aactaacata catcatagtt actaaactat ggtaactata
     4561 attttattaa ctatattaag cactgattag tactataact caatataagc atatcccctg
     4621 tatcgtaact agagaaccaa acgacggaaa aagcgatata gataagttta gatagtgcac
     4681 caatttttta cttttctatt ttagaatcta gcatttccaa tgcttactaa tactattttt
     4741 gatttttaac ttgttttaaa aaagcaccta ctaattagga ttctagctct atattctagt
     4801 tatacattct ctatattcta gctctatatt ctagttatac attctctata ttctagctct
     4861 atattctagt tatacattct ctatattcta gctctatatt ctagttatac attctctata
     4921 ttctagctct atattctagt tatacattct ctatattcta gctctatatt ctagttatac
     4981 attctctata ttctagctct atattctagt tatacattct ctaaaaaaca tatatatata
     5041 tactacatat atatgtacaa ttattttacc aaaagaaatt tttaaaacgg cttaatttca
     5101 atactttgat gccatttttt ttttgcaatt ctcgtacaat tattttacca aaagaaattt
     5161 ttattaatta gtttttttgc ataaaaatag gactagaaaa tcaattctaa tcctatcaaa
     5221 aatctacttg tttaatctat atgtatagta cataaatcct ctatcattta ttttagtttc
     5281 ccatacatct aaaattttag tatctaataa agattctatc atatttgaaa ttctttgctg
     5341 tgttactcct aaatcttttg ctaattcctc ttgattaatt cttaatatat ttccttttat
     5401 tttatcatct ttgaccaata aatcttgctt atatctcata tgagtatata atcttaattc
     5461 ttcagttgaa attattcccc aaattactgc catggtaaca aagtaactaa tagtatattg
     5521 tttatctata ttacatctaa cttcttttac attgtatgta tcttttattc ctagctttgt
     5581 aacacctttt gtttttgtaa tataatcatt atgttgaagt tcttttaatg tttctcttaa
     5641 tgtcttttca ctcataacaa tgtttttaac tttcttctta cgtttatagg ttatcagctc
     5701 cattatatcg tctatgttta attctctatc tttattgtta agtaacacat tatagataaa
     5761 caattgatta ccagtcatta ttttaacccc tttcctattc ttatatttca aatcctttga
     5821 gtgcttatgt ggcatattga acaaagtatc ttcatcacta tatataaaat ctgattctat
     5881 tttattccaa caatcactat taaactcaca acctgtacaa ccaaaattaa ccgttttaag
     5941 cttctcatac atatacttga cttgatattc tagctcacta tttgaaagac tattttgtga
     6001 tatatagttc cattctctag caacggatat tacttgactt aaacttttat tcctcaatct
     6061 taaagtcaca actatttttt gcaaatctag gtttttatgt ccatctttac taccattttt
     6121 taaaatatct actatacaat ttggtagctt actatctaat atatattttg tattagtatt
     6181 acctactgtt tttacatctt tgcaatattt tttagctaac ttttctatgt catatctata
     6241 tatatttttg tcttggtgta ttatttttac ttgtttagta gtatttttaa tattatatgt
     6301 ataaggtact cttaatactt gagtagataa atttgcttgt ttatctgcac ccaatttatt
     6361 tattaatgta tttgtaactt ctgatactaa cttaatgtta ttagttttat taattagcac
     6421 ataaacatgg aatccatttc cactatcaac taggatatga taatgtaatc ctatcttagt
     6481 aaataaatta attatatctt tataattaaa accttgtcct aattcttttt tgtcaaaatc
     6541 aaatcctaat gcatatctat ttattaagtt ttcattggtt cctgcttttc catcagttgt
     6601 acataggtta aaaaatgtat tttcataata aatatttttt gcgaaggtaa attctactat
     6661 atcatctata tttttaaaaa aagttgtttt actatattct ttaaattctt ctccgagttc
     6721 taattgttta ctattacttt tattattttg aaatactcta acatattcat tatcaataag
     6781 attatttcct tgtcctatac catctctaaa tactgccatt agaaattttt tcaacttata
     6841 ttttgaatct aattgttcca tatatttatt atctccttat ctagtttttt taagagataa
     6901 cactacatta aatatatact tattgactaa tcaactttta caactattga ataattttag
     6961 cttagtatgt ataataatat tgttaagatg ctaaaattga tagcaatatt attatataac
     7021 tacagatact taatcagtta gaccgatatt ggagttggta gctccataaa ctcaattggt
     7081 agttgagttg tcattgtcta gtagtattat ctcattttgt tgttatctta aaatattatt
     7141 ttaaaaatac tctgcttggt cggcttaggg tatttttttt gtttaattcc aatattggtt
     7201 caaactcctg tgtaatactt ttgacacttc catctgtatt gaattgacag ttgatatggt
     7261 atctcattat tggcttgttc ttatgttttt ctatgtattt attttttact ttcaaataat
     7321 ttgtatttcc atttttagtt atgtttaaca cactatcctt attactcaat aaaattattt
     7381 tttgttttaa catatgattt atagaatcta aaaattgata ttgacttata atagtacttt
     7441 ctacctctct agctttttct agtgtttcaa attcttcaaa ctctgctata ttgctatagt
     7501 atagcttatc attaataata ttatctgaca actttaattc cattttataa aacaattcca
     7561 agttatgtat aaaattttta atgttacaac atatagactt attatttgtt cttatatggt
     7621 atctccattt actacttcta gctctcatgt tatatgcaca actactccca acataaacat
     7681 atattgcttg ttctaactca tatttttgta aaatttcaac atcatatcca gtcaaatcgt
     7741 actctctctt atatacaaat ggtttttcta attttttgtt caataaatct cctcctattt
     7801 cttacttaat ttagctaaaa atctatttat atttcacctc ctttttgact ttaagcctac
     7861 gaataccata tttatgatta taacatcttc tatttttccc aaatccttac atctccccta
     7921 atttttagac ttaagggcga attctgcatc aa
//
