LOCUS       V003828                 7922 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003828
VERSION     V003828
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7922)
  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. 196 (12), 2290-2300 (2014)
   PUBMED   24727226
REFERENCE   2  (bases 1 to 7922)
  AUTHORS   Ransom EM, Weiss DS, Ellermeier CD.
  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 7922)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7922)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "J.
            Bacteriol."; date: "2014"; volume: "196"; issue: "12"; pages:
            "2290-2300"
            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..7922
                     /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..1589
                     /label="ECFP"
                     /note="enhanced CFP"
     CDS             complement(1941..2309)
                     /label="traJ"
                     /note="oriT-recognizing protein"
     oriT            complement(2342..2451)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             complement(2830..3450)
                     /gene="catP"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from Clostridium
                     perfringens. Accession#: P26826"
     rep_origin      complement(3813..4401)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5194..6831)
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label="repA"
                     /note="plasmid replication; derived from Clostridium
                     difficile"
                     /protein_id="AMA07794.1"
                     /translation="MEQLDSKYKLKKFLMAVFRDGIGQGNNLIDNEYVRVFQNNKSNSK
                     QLELGEEFKEYSKTTFFKNIDDIVEFTFAKNIYYENTFFNLCTTDGKAGTNENLINRYA
                     LGFDFDKKELGQGFNYKDIINLFTKIGLHYHILVDSGNGFHVYVLINKTNNIKLVSEVT
                     NTLINKLGADKQANLSTQVLRVPYTYNIKNTTKQVKIIHQDKNIYRYDIEKLAKKYCKD
                     VKTVGNTNTKYILDSKLPNCIVDILKNGSKDGHKNLDLQKIVVTLRLRNKSLSQVISVA
                     REWNYISQNSLSNSELEYQVKYMYEKLKTVNFGCTGCEFNSDCWNKIESDFIYSDEDTL
                     FNMPHKHSKDLKYKNRKGVKIMTGNQLFIYNVLLNNKDRELNIDDIMELITYKRKKKVK
                     NIVMSEKTLRETLKELQHNDYITKTKGVTKLGIKDTYNVKEVRCNIDKQYTISYFVTMA
                     VIWGIISTEELRLYTHMRYKQDLLVKDDKIKGNILRINQEELAKDLGVTQQRISNMIES
                     LLDTKILDVWETKINDRGFMYYTYRLNK"
     gene            complement(5194..6831)
                     /gene="repA"
                     /label="repA"
     CDS             complement(7106..7618)
                     /codon_start=1
                     /gene="orfB"
                     /product="OrfB"
                     /label="orfB"
                     /protein_id="AMA07792.1"
                     /translation="MRARSSKWRYHIRTNNKSICCNIKNFIHNLELFYKMELKLSDNII
                     NDKLYYSNIAEFEEFETLEKAREVESTIISQYQFLDSINHMLKQKIILLSNKDSVLNIT
                     KNGNTNYLKVKNKYIEKHKNKPIMRYHINCQFNTDGSVKSITQEFEPILELNKKNTLSR
                     PSRVFLK"
     gene            complement(7106..7618)
                     /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 ttatggtttc aaaaggagaa gaattattta
      901 caggggtagt accaatttta gtagaacttg acggagatgt aaatggacac aaattttcag
      961 tatcaggaga aggagaagga gatgcaacat acggaaaatt aacacttaaa ttcatttgta
     1021 caacaggtaa actaccggtt ccatggccca cacttgtgac tactttaaca tggggagtac
     1081 aatgtttttc aagatatcca gatcatatga aacaacacga cttttttaaa tcagcaatgc
     1141 ctgaaggata cgtacaagaa agaacaatat tttttaaaga tgatggtaac tataaaacga
     1201 gagcagaagt aaaatttgaa ggagatacat tagtaaacag aattgaatta aaaggaatag
     1261 actttaaaga agatggaaat atcctaggac ataaattaga atacaattac atcagtcata
     1321 acgtatacat aacggcagat aaacagaaaa atggaattaa agctaacttt aaaataagac
     1381 acaatataga agacggatca gtacaattag cagatcacta tcaacaaaat acacctatcg
     1441 gagatggccc agtgcttcta ccagataatc attacttaag cacgcaaagc gctttatcta
     1501 aagacccaaa tgaaaaaaga gaccacatgg tattattaga attcgtaaca gcagcaggaa
     1561 ttacactagg aatggatgaa ttatataagt aaggatccta taagttttaa taaaacttta
     1621 aatagaaaaa ggcttctctc atgagaagtc ttttttattt aaaataaata taaaataaaa
     1681 tagaggctat aaatagcctc tattttatgt gagaaatccc taaataaaaa gatgccagtg
     1741 tgtgctggaa ttcgccctta ggctgcgcta gggacctctt tagctccttg gaagctgtca
     1801 gtagtagggg atcggtcttg ccttgctcgt cggtgatgta cttcaccagc tccgcgaagt
     1861 cgctcttctt gatggagcgc atggggacgt gcttggcaat cacgcgcacc ccccggccgt
     1921 tttagcggct aaaaaagtca tggctctgcc ctcgggcgga ccacgcccat catgaccttg
     1981 ccaagctcgt cctgcttctc ttcgatcttc gccagcaggg cgaggatcgt ggcatcaccg
     2041 aaccgcgccg tgcgcgggtc gtcggtgagc cagagtttca gcaggccgcc caggcggccc
     2101 aggtcgccat tgatgcgggc cagctcgcgg acgtgctcat agtccacgac gcccgtgatt
     2161 ttgtagccct ggccgacggc cagcaggtag gccgacaggc tcatgccggc cgccgccgcc
     2221 ttttcctcaa tcgctcttcg ttcgtctgga aggcagtaca ccttgatagg tgggctgccc
     2281 ttcctggttg gcttggtttc atcagccatc cgcttgccct catctgttac gccggcggta
     2341 gccggccagc ctcgcagagc aggattcccg ttgagcaccg ccaggtgcga ataagggaca
     2401 gtgaagaagg aacacccgct cgcgggtggg cctacttcac ctatcctgcc cggctgacgc
     2461 cgttggatac accaaggaaa gtctacacga accctttggc aaaatcctgt atatcgtgcg
     2521 aaaaaggatg gatataccga aaaaatcgct ataatgaccc cgaagcagga agggcgaatt
     2581 ctgcagatct gaccggtctc tgaaaatata aaaaccacag attgatacta aaaccttggt
     2641 tgtgttgctt ttcggggctt aaatcaagga aaaatccttg ttttaagcct ttcaaaaaga
     2701 aacacaaggt ctttgtacta acctgtggtt atgtataaaa ttgtagattt tagggtaaca
     2761 aaaaacaccg tatttctacg atgtttttgc ttaaatactt gtttttagtt acagacaaac
     2821 ctgaagttaa ctatttatca attcctgcaa ttcgtttaca aaacggcaaa tgtgaaatcc
     2881 gtcacatact gcgtgatgaa cttgaattgc caaaggaagt ataattttgt tatcttcttt
     2941 ataatatttc cccatagtaa aaataggaat caaataatca tatcctttct gcaaattcag
     3001 attaaagcca tcgaaggttg accacggtat catagataca ttaaaaatgt tttccggagc
     3061 atttggcttt ccttccattc tatgattgtt tccataccgt tgcgtatcac tttcataatc
     3121 tgctaaaaat gatttaaagt cagacttaca ctcagtccaa aggctggaaa atgtttcagt
     3181 atcattgtga aatattgtat agcttggtat catctcatca tatatcccca attcaccatc
     3241 ttgattgatt gccgtcctaa actctgaatg gcggtttaca atcattgcaa tataataaag
     3301 cattgcagga tatagtttca ttcccttttc ctttatttgt gtgatatcca ctttaacggt
     3361 catgctgtag gtacaaggta cacttgcaaa gtagtggtca aaatactctt ttctgttcca
     3421 actattttta tcaatttttt caaataccat ctaagttccc tctcaaattc aagtttatcg
     3481 ctctaatgaa caaagatatt ataccacatt tttgtgaatt tttcaacttg cccacttcga
     3541 ctgcactccc gacttaataa cttcttgaac acttgccgaa aaagaaaaac tgccgggtac
     3601 gtacccgggg atcgatcccc ggccgagcgc tcttccgctt cctcgctcac tgactcgctg
     3661 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3721 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3781 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3841 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3901 caggcgtttc ccccttgaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3961 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4021 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4081 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4141 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4201 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4261 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4321 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4381 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4441 tggaacgaaa actcacgtta agggattttg gtcatgaaat gcaagtttct aactaacata
     4501 catcatagtt actaaactat ggtaactata attttattaa ctatattaag cactgattag
     4561 tactataact caatataagc atatcccctg tatcgtaact agagaaccaa acgacggaaa
     4621 aagcgatata gataagttta gatagtgcac caatttttta cttttctatt ttagaatcta
     4681 gcatttccaa tgcttactaa tactattttt gatttttaac ttgttttaaa aaagcaccta
     4741 ctaattagga ttctagctct atattctagt tatacattct ctatattcta gctctatatt
     4801 ctagttatac attctctata ttctagctct atattctagt tatacattct ctatattcta
     4861 gctctatatt ctagttatac attctctata ttctagctct atattctagt tatacattct
     4921 ctatattcta gctctatatt ctagttatac attctctata ttctagctct atattctagt
     4981 tatacattct ctaaaaaaca tatatatata tactacatat atatgtacaa ttattttacc
     5041 aaaagaaatt tttaaaacgg cttaatttca atactttgat gccatttttt ttttgcaatt
     5101 ctcgtacaat tattttacca aaagaaattt ttattaatta gtttttttgc ataaaaatag
     5161 gactagaaaa tcaattctaa tcctatcaaa aatctacttg tttaatctat atgtatagta
     5221 cataaatcct ctatcattta ttttagtttc ccatacatct aaaattttag tatctaataa
     5281 agattctatc atatttgaaa ttctttgctg tgttactcct aaatcttttg ctaattcctc
     5341 ttgattaatt cttaatatat ttccttttat tttatcatct ttgaccaata aatcttgctt
     5401 atatctcata tgagtatata atcttaattc ttcagttgaa attattcccc aaattactgc
     5461 catggtaaca aagtaactaa tagtatattg tttatctata ttacatctaa cttcttttac
     5521 attgtatgta tcttttattc ctagctttgt aacacctttt gtttttgtaa tataatcatt
     5581 atgttgaagt tcttttaatg tttctcttaa tgtcttttca ctcataacaa tgtttttaac
     5641 tttcttctta cgtttatagg ttatcagctc cattatatcg tctatgttta attctctatc
     5701 tttattgtta agtaacacat tatagataaa caattgatta ccagtcatta ttttaacccc
     5761 tttcctattc ttatatttca aatcctttga gtgcttatgt ggcatattga acaaagtatc
     5821 ttcatcacta tatataaaat ctgattctat tttattccaa caatcactat taaactcaca
     5881 acctgtacaa ccaaaattaa ccgttttaag cttctcatac atatacttga cttgatattc
     5941 tagctcacta tttgaaagac tattttgtga tatatagttc cattctctag caacggatat
     6001 tacttgactt aaacttttat tcctcaatct taaagtcaca actatttttt gcaaatctag
     6061 gtttttatgt ccatctttac taccattttt taaaatatct actatacaat ttggtagctt
     6121 actatctaat atatattttg tattagtatt acctactgtt tttacatctt tgcaatattt
     6181 tttagctaac ttttctatgt catatctata tatatttttg tcttggtgta ttatttttac
     6241 ttgtttagta gtatttttaa tattatatgt ataaggtact cttaatactt gagtagataa
     6301 atttgcttgt ttatctgcac ccaatttatt tattaatgta tttgtaactt ctgatactaa
     6361 cttaatgtta ttagttttat taattagcac ataaacatgg aatccatttc cactatcaac
     6421 taggatatga taatgtaatc ctatcttagt aaataaatta attatatctt tataattaaa
     6481 accttgtcct aattcttttt tgtcaaaatc aaatcctaat gcatatctat ttattaagtt
     6541 ttcattggtt cctgcttttc catcagttgt acataggtta aaaaatgtat tttcataata
     6601 aatatttttt gcgaaggtaa attctactat atcatctata tttttaaaaa aagttgtttt
     6661 actatattct ttaaattctt ctccgagttc taattgttta ctattacttt tattattttg
     6721 aaatactcta acatattcat tatcaataag attatttcct tgtcctatac catctctaaa
     6781 tactgccatt agaaattttt tcaacttata ttttgaatct aattgttcca tatatttatt
     6841 atctccttat ctagtttttt taagagataa cactacatta aatatatact tattgactaa
     6901 tcaactttta caactattga ataattttag cttagtatgt ataataatat tgttaagatg
     6961 ctaaaattga tagcaatatt attatataac tacagatact taatcagtta gaccgatatt
     7021 ggagttggta gctccataaa ctcaattggt agttgagttg tcattgtcta gtagtattat
     7081 ctcattttgt tgttatctta aaatattatt ttaaaaatac tctgcttggt cggcttaggg
     7141 tatttttttt gtttaattcc aatattggtt caaactcctg tgtaatactt ttgacacttc
     7201 catctgtatt gaattgacag ttgatatggt atctcattat tggcttgttc ttatgttttt
     7261 ctatgtattt attttttact ttcaaataat ttgtatttcc atttttagtt atgtttaaca
     7321 cactatcctt attactcaat aaaattattt tttgttttaa catatgattt atagaatcta
     7381 aaaattgata ttgacttata atagtacttt ctacctctct agctttttct agtgtttcaa
     7441 attcttcaaa ctctgctata ttgctatagt atagcttatc attaataata ttatctgaca
     7501 actttaattc cattttataa aacaattcca agttatgtat aaaattttta atgttacaac
     7561 atatagactt attatttgtt cttatatggt atctccattt actacttcta gctctcatgt
     7621 tatatgcaca actactccca acataaacat atattgcttg ttctaactca tatttttgta
     7681 aaatttcaac atcatatcca gtcaaatcgt actctctctt atatacaaat ggtttttcta
     7741 attttttgtt caataaatct cctcctattt cttacttaat ttagctaaaa atctatttat
     7801 atttcacctc ctttttgact ttaagcctac gaataccata tttatgatta taacatcttc
     7861 tatttttccc aaatccttac atctccccta atttttagac ttaagggcga attctgcatc
     7921 aa
//
