LOCUS       V004419                 7869 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004419
VERSION     V004419
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7869)
  AUTHORS   Heap JT, Kuehne SA, Ehsaan M, Cartman ST, Cooksley CM, Scott JC,
            Minton NP.
  TITLE     The ClosTron: Mutagenesis in Clostridium refined and streamlined
  JOURNAL   J. Microbiol. Methods 80 (1), 49-55 (2010)
   PUBMED   19891996
REFERENCE   2  (bases 1 to 7869)
  AUTHORS   Heap JT, Kuehne KA, Ehsaan M, Cartman ST, Cooksley CM, Minton NP.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-SEP-2010) School of Molecular Medical Sciences, The
            University of Nottingham, Centre for Biomolecular Sciences,
            Nottingham, Nottinghamshire NG72RD, UK
REFERENCE   3  (bases 1 to 7869)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7869)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J.
            Microbiol. Methods"; date: "2010"; volume: "80"; issue: "1"; pages:
            "49-55"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (28-SEP-2010) School of Molecular Medical Sciences, The University
            of Nottingham, Centre for Biomolecular Sciences, Nottingham,
            Nottinghamshire NG72RD, UK"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7869
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             239..412
                     /label="lacZ-alpha"
                     /note="LacZ-alpha fragment of beta-galactosidase"
     RBS             1071..1093
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1285..3081
                     /gene="ltrA"
                     /label="Group II intron-encoded protein LtrA"
                     /note="Group II intron-encoded protein LtrA from
                     Lactococcus lactis subsp. cremoris (strain MG1363).
                     Accession#: P0A3U1"
     terminator      3269..3355
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3447..3474
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     rep_origin      6319..6907
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     oriT            7228..7337
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             7370..7738
                     /label="traJ"
                     /note="oriT-recognizing protein"
ORIGIN
        1 gggccccctg cagggtgtag tagcctgtga aataagtaag gaaaaaaaag aagtaagtgt
       61 tatatatgat gattattttg tagatgtaga taggataata gaatccatag aaaatatagg
      121 ttatacagtt atataaaaat tactttaaaa attaataaaa acatggtaaa atataaatcg
      181 tataaagttg tgtaattttt aagcttgagc tcataacaat ttcacacagg aaacagctat
      241 gaccatgatt acggattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
      301 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
      361 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcta
      421 ataaagatct tgtacaatct gtaggagaac ctatgggaac gaaacgaaag cgatgccgag
      481 aatctgaatt taccaagact taacactaac tggggatacc ctaaacaaga atgcctaata
      541 gaaaggagga aaaaggctat agcactagag cttgaaaatc ttgcaagggt acggagtact
      601 cgtagtagtc tgagaagggt aacgcccttt acatggcaaa ggggtacagt tattgtgtac
      661 taaaattaaa aattgattag ggaggaaaac ctcaaaatga aaccaacaat ggcaatttta
      721 gaaagaatca gtaaaaattc acaagaaaat atagacgaag tttttacaag actttatcgt
      781 tatcttttac gtccagatat ttattacgtg gcgacgcgtt gggaaatggc aatgatagcg
      841 aaacaacgta aaactcttgt tgtatgcttt cattgtcatc gtcacgtgat tcataaacac
      901 aagtgaatgt cgacagtgaa tttttacgaa cgaacaataa cagagccgta tactccgaga
      961 ggggtacgta cggttcccga agagggtggt gcaaaccagt cacagtaatg tgaacaaggc
     1021 ggtacctccc tacttcacca tatcattttc tgcagccccc tagaaataat tttgtttaac
     1081 tttaagaagg agatatacat atatggctag atcgtccatt ccgacagcat cgccagtcac
     1141 tatggcgtgc tgctagcgct atatgcgttg atgcaatttc tatgcactcg tagtagtctg
     1201 agaagggtaa cgccctttac atggcaaagg ggtacagtta ttgtgtacta aaattaaaaa
     1261 ttgattaggg aggaaaacct caaaatgaaa ccaacaatgg caattttaga aagaatcagt
     1321 aaaaattcac aagaaaatat agacgaagtt tttacaagac tttatcgtta tcttttacgt
     1381 ccagatattt attacgtggc gtatcaaaat ttatattcca ataaaggagc ttccacaaaa
     1441 ggaatattag atgatacagc ggatggcttt agtgaagaaa aaataaaaaa gattattcaa
     1501 tctttaaaag acggaactta ctatcctcaa cctgtacgaa gaatgtatat tgcaaaaaag
     1561 aattctaaaa agatgagacc tttaggaatt ccaactttca cagataaatt gatccaagaa
     1621 gctgtgagaa taattcttga atctatctat gaaccggtat tcgaagatgt gtctcacggt
     1681 tttagacctc aacgaagctg tcacacagct ttgaaaacaa tcaaaagaga gtttggcggc
     1741 gcaagatggt ttgtggaggg agatataaaa ggctgcttcg ataatataga ccacgttaca
     1801 ctcattggac tcatcaatct taaaatcaaa gatatgaaaa tgagccaatt gatttataaa
     1861 tttctaaaag caggttatct ggaaaactgg cagtatcaca aaacttacag cggaacacct
     1921 caaggtggaa ttctatctcc tcttttggcc aacatctatc ttcatgaatt ggataagttt
     1981 gttttacaac tcaaaatgaa gtttgaccga gaaagtccag aaagaataac acctgaatat
     2041 cgggagctcc acaatgagat aaaaagaatt tctcaccgtc tcaagaagtt ggagggtgaa
     2101 gaaaaagcta aagttctttt agaatatcaa gaaaaacgta aaagattacc cacactcccc
     2161 tgtacctcac agacaaataa agtattgaaa tacgtccggt atgcggacga cttcattatc
     2221 tctgttaaag gaagcaaaga ggactgtcaa tggataaaag aacaattaaa actttttatt
     2281 cataacaagc taaaaatgga attgagtgaa gaaaaaacac tcatcacaca tagcagtcaa
     2341 cccgctcgtt ttctgggata tgatatacga gtaaggagat ctggaacgat aaaacgatct
     2401 ggtaaagtca aaaagagaac actcaatggg agtgtagaac tccttattcc tcttcaagac
     2461 aaaattcgtc aatttatttt tgacaagaaa atagctatcc aaaagaaaga tagctcatgg
     2521 tttccagttc acaggaaata tcttattcgt tcaacagact tagaaatcat cacaatttat
     2581 aattctgaac tccgcgggat ttgtaattac tacggtctag caagtaattt taaccagctc
     2641 aattattttg cttatcttat ggaatacagc tgtctaaaaa cgatagcctc caaacataag
     2701 ggaacacttt caaaaaccat ttccatgttt aaagatggaa gtggttcgtg ggggatcccg
     2761 tatgagataa agcaaggtaa gcagcgccgt tattttgcaa attttagtga atgtaaatcc
     2821 ccttatcaat ttacggatga gataagtcaa gctcctgtat tgtatggcta tgcccggaat
     2881 actcttgaaa acaggttaaa agctaaatgt tgtgaattat gtgggacgtc tgatgaaaat
     2941 acttcctatg aaattcacca tgtcaataag gtcaaaaatc ttaaaggcaa agaaaaatgg
     3001 gaaatggcaa tgatagcgaa acaacgtaaa actcttgttg tatgctttca ttgtcatcgt
     3061 cacgtgattc ataaacacaa gtgaatgtcg agcacccgtt ctcggagcac tgtccgaccg
     3121 ctttggccgc cgcccagtcc tgctcgcttc gctacttgga gccactatcg actacgcgat
     3181 catggcgacc acacccgtcc tgtggatcgc caagccgccg atggtagtgt ggggtctccc
     3241 catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg
     3301 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc
     3361 gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc
     3421 ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt
     3481 tctacaaact cttcctgtcg tcatatctac aagccatccc cccacagata cgggcgcgcc
     3541 gccattattt ttttgaacaa ttgacaattc atttcttatt ttttattaag tgatagtcaa
     3601 aaggcataac agtgctgaat agaaagaaat ttacagaaaa gaaaattata gaatttagta
     3661 tgattaatta tactcattta tgaatgttta attgaataca aaaaaaaata cttgttatgt
     3721 attcaattac gggttaaaat atagacaagt tgaaaaattt aataaaaaaa taagtcctca
     3781 gctcttatat attaagctac caacttagta tataagccaa aacttaaatg tgctaccaac
     3841 acatcaagcc gttagagaac tctatctata gcaatatttc aaatgtaccg acatacaaga
     3901 gaaacattaa ctatatatat tcaatttatg agattatctt aacagatata aatgtaaatt
     3961 gcaataagta agatttagaa gtttatagcc tttgtgtatt ggaagcagta cgcaaaggct
     4021 tttttatttg ataaaaatta gaagtatatt tattttttca taattaattt atgaaaatga
     4081 aagggggtga gcaaagtgac agaggaaagc agtatcttat caaataacaa ggtattagca
     4141 atatcattat tgactttagc agtaaacatt atgactttta tagtgcttgt agctaagtag
     4201 tacgaaaggg ggagctttaa aaagctcctt ggaatacata gaattcataa attaatttat
     4261 gaaaagaagg gcgtatatga aaacttgtaa aaattgcaaa gagtttatta aagatactga
     4321 aatatgcaaa atacattcgt tgatgattca tgataaaaca gtagcaacct attgcagtaa
     4381 atacaatgag tcaagatgtt tacataaagg gaaagtccaa tgtattaatt gttcaaagat
     4441 gaaccgatat ggatggtgtg ccataaaaat gagatgtttt acagaggaag aacagaaaaa
     4501 agaacgtaca tgcattaaat attatgcaag gagctttaaa aaagctcatg taaagaagag
     4561 taaaaagaaa aaataattta tttattaatt taatattgag agtgccgaca cagtatgcac
     4621 taaaaaatat atctgtggtg tagtgagccg atacaaaagg atagtcactc gcattttcat
     4681 aatacatctt atgttatgat tatgtgtcgg tgggacttca cgacgaaaac ccacaataaa
     4741 aaaagagttc ggggtagggt taagcatagt tgaggcaact aaacaatcaa gctaggatat
     4801 gcagtagcag accgtaaggt cgttgtttag gtgtgttgta atacatacgc tattaagatg
     4861 taaaaatacg gataccaatg aagggaaaag tataattttt ggatgtagtt tgtttgttca
     4921 tctatgggca aactacgtcc aaagccgttt ccaaatctgc taaaaagtat atcctttcta
     4981 aaatcaaagt caagtatgaa atcataaata aagtttaatt ttgaagttat tatgatatta
     5041 tgtttttcta ttaaaataaa ttaagtatat agaatagttt aataatagta tatacttaat
     5101 gtgataagtg tctgacagtg tcacagaaag gatgattgtt atggattata agcggccggc
     5161 ccaatgaata ggtttacact tactttagtt ttatggaaat gaaagatcat atcatatata
     5221 atctagaata aaattaacta aaataattat tatctagata aaaaatttag aagccaatga
     5281 aatctataaa taaactaaat taagtttatt taattaacaa ctatggatat aaaataggta
     5341 ctaatcaaaa tagtgaggag gatatatttg aatacatacg aacaaattaa taaagtgaaa
     5401 aaaatacttc ggaaacattt aaaaaataac cttattggta cttacatgtt tggatcagga
     5461 gttgagagtg gactaaaacc aaatagtgat cttgactttt tagtcgtcgt atctgaacca
     5521 ttgacagatc aaagtaaaga aatacttata caaaaaatta gacctatttc aaagaaaata
     5581 ggagataaaa gcaacttacg atatattgaa ttaacaatta ttattcagca agaaatggta
     5641 ccgtggaatc atcctcccaa acaagaattt atttatggag aatggttaca agagctttat
     5701 gaacaaggat acattcctca gaaggaatta aattcagatt taaccataat gctttaccaa
     5761 gcaaaacgaa aaaataaaag aatatacgga aattatgact tagaggaatt actacctgat
     5821 attccatttt ctgatgtgag aagagccatt atggattcgt cagaggaatt aatagataat
     5881 tatcaggatg atgaaaccaa ctctatatta actttatgcc gtatgatttt aactatggac
     5941 acgggtaaaa tcataccaaa agatattgcg ggaaatgcag tggctgaatc ttctccatta
     6001 gaacataggg agagaatttt gttagcagtt cgtagttatc ttggagagaa tattgaatgg
     6061 actaatgaaa atgtaaattt aactataaac tatttaaata acagattaaa aaaattataa
     6121 aaaaattgaa aaaatggtgg aaacactttt ttcaattttt ttgttttatt atttaatatt
     6181 tgggaaatat tcattctaat tggtaatcag attttagaag tttaaactcc tttttgataa
     6241 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6301 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6361 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6421 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     6481 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6541 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6601 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6661 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6721 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6781 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6841 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6901 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6961 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     7021 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     7081 agcggaagag cgcccaatac gcagggcccc ctgcttcggg gtcattatag cgattttttc
     7141 ggtatatcca tcctttttcg cacgatatac aggattttgc caaagggttc gtgtagactt
     7201 tccttggtgt atccaacggc gtcagccggg caggataggt gaagtaggcc cacccgcgag
     7261 cgggtgttcc ttcttcactg tcccttattc gcacctggcg gtgctcaacg ggaatcctgc
     7321 tctgcgaggc tggccggcta ccgccggcgt aacagatgag ggcaagcgga tggctgatga
     7381 aaccaagcca accaggaagg gcagcccacc tatcaaggtg tactgccttc cagacgaacg
     7441 aagagcgatt gaggaaaagg cggcggcggc cggcatgagc ctgtcggcct acctgctggc
     7501 cgtcggccag ggctacaaaa tcacgggcgt cgtggactat gagcacgtcc gcgagctggc
     7561 ccgcatcaat ggcgacctgg gccgcctggg cggcctgctg aaactctggc tcaccgacga
     7621 cccgcgcacg gcgcggttcg gtgatgccac gatcctcgcc ctgctggcga agatcgaaga
     7681 gaagcaggac gagcttggca aggtcatgat gggcgtggtc cgcccgaggg cagagccatg
     7741 acttttttag ccgctaaaac ggccgggggg tgcgcgtgat tgccaagcac gtccccatgc
     7801 gctccatcaa gaagagcgac ttcgcggagc tggtgaagta catcaccgac gagcaaggca
     7861 agaccgatc
//
