LOCUS       V004422                 7711 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004422
VERSION     V004422
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7711)
  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 7711)
  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 7711)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7711)
  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..7711
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             234..407
                     /label="lacZ-alpha"
                     /note="LacZ-alpha fragment of beta-galactosidase"
     RBS             1066..1088
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1280..3076
                     /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      3264..3350
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3442..3469
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     CDS             5265..5999
                     /gene="ermBP"
                     /label="rRNA adenine N-6-methyltransferase"
                     /note="rRNA adenine N-6-methyltransferase from Enterococcus
                     faecalis. Accession#: P0A4D5"
     rep_origin      6156..6744
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     oriT            7065..7174
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             7207..7575
                     /label="traJ"
                     /note="oriT-recognizing protein"
ORIGIN
        1 ccctgcaggg tgtagtagcc tgtgaaataa gtaaggaaaa aaaagaagta agtgttatat
       61 atgatgatta ttttgtagat gtagatagga taatagaatc catagaaaat ataggttata
      121 cagttatata aaaattactt taaaaattaa taaaaacatg gtaaaatata aatcgtataa
      181 agttgtgtaa tttttaagct tgagctcata acaatttcac acaggaaaca gctatgacca
      241 tgattacgga ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
      301 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
      361 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgctaataaa
      421 gatcttgtac aatctgtagg agaacctatg ggaacgaaac gaaagcgatg ccgagaatct
      481 gaatttacca agacttaaca ctaactgggg ataccctaaa caagaatgcc taatagaaag
      541 gaggaaaaag gctatagcac tagagcttga aaatcttgca agggtacgga gtactcgtag
      601 tagtctgaga agggtaacgc cctttacatg gcaaaggggt acagttattg tgtactaaaa
      661 ttaaaaattg attagggagg aaaacctcaa aatgaaacca acaatggcaa ttttagaaag
      721 aatcagtaaa aattcacaag aaaatataga cgaagttttt acaagacttt atcgttatct
      781 tttacgtcca gatatttatt acgtggcgac gcgttgggaa atggcaatga tagcgaaaca
      841 acgtaaaact cttgttgtat gctttcattg tcatcgtcac gtgattcata aacacaagtg
      901 aatgtcgaca gtgaattttt acgaacgaac aataacagag ccgtatactc cgagaggggt
      961 acgtacggtt cccgaagagg gtggtgcaaa ccagtcacag taatgtgaac aaggcggtac
     1021 ctccctactt caccatatca ttttctgcag ccccctagaa ataattttgt ttaactttaa
     1081 gaaggagata tacatatatg gctagatcgt ccattccgac agcatcgcca gtcactatgg
     1141 cgtgctgcta gcgctatatg cgttgatgca atttctatgc actcgtagta gtctgagaag
     1201 ggtaacgccc tttacatggc aaaggggtac agttattgtg tactaaaatt aaaaattgat
     1261 tagggaggaa aacctcaaaa tgaaaccaac aatggcaatt ttagaaagaa tcagtaaaaa
     1321 ttcacaagaa aatatagacg aagtttttac aagactttat cgttatcttt tacgtccaga
     1381 tatttattac gtggcgtatc aaaatttata ttccaataaa ggagcttcca caaaaggaat
     1441 attagatgat acagcggatg gctttagtga agaaaaaata aaaaagatta ttcaatcttt
     1501 aaaagacgga acttactatc ctcaacctgt acgaagaatg tatattgcaa aaaagaattc
     1561 taaaaagatg agacctttag gaattccaac tttcacagat aaattgatcc aagaagctgt
     1621 gagaataatt cttgaatcta tctatgaacc ggtattcgaa gatgtgtctc acggttttag
     1681 acctcaacga agctgtcaca cagctttgaa aacaatcaaa agagagtttg gcggcgcaag
     1741 atggtttgtg gagggagata taaaaggctg cttcgataat atagaccacg ttacactcat
     1801 tggactcatc aatcttaaaa tcaaagatat gaaaatgagc caattgattt ataaatttct
     1861 aaaagcaggt tatctggaaa actggcagta tcacaaaact tacagcggaa cacctcaagg
     1921 tggaattcta tctcctcttt tggccaacat ctatcttcat gaattggata agtttgtttt
     1981 acaactcaaa atgaagtttg accgagaaag tccagaaaga ataacacctg aatatcggga
     2041 gctccacaat gagataaaaa gaatttctca ccgtctcaag aagttggagg gtgaagaaaa
     2101 agctaaagtt cttttagaat atcaagaaaa acgtaaaaga ttacccacac tcccctgtac
     2161 ctcacagaca aataaagtat tgaaatacgt ccggtatgcg gacgacttca ttatctctgt
     2221 taaaggaagc aaagaggact gtcaatggat aaaagaacaa ttaaaacttt ttattcataa
     2281 caagctaaaa atggaattga gtgaagaaaa aacactcatc acacatagca gtcaacccgc
     2341 tcgttttctg ggatatgata tacgagtaag gagatctgga acgataaaac gatctggtaa
     2401 agtcaaaaag agaacactca atgggagtgt agaactcctt attcctcttc aagacaaaat
     2461 tcgtcaattt atttttgaca agaaaatagc tatccaaaag aaagatagct catggtttcc
     2521 agttcacagg aaatatctta ttcgttcaac agacttagaa atcatcacaa tttataattc
     2581 tgaactccgc gggatttgta attactacgg tctagcaagt aattttaacc agctcaatta
     2641 ttttgcttat cttatggaat acagctgtct aaaaacgata gcctccaaac ataagggaac
     2701 actttcaaaa accatttcca tgtttaaaga tggaagtggt tcgtggggga tcccgtatga
     2761 gataaagcaa ggtaagcagc gccgttattt tgcaaatttt agtgaatgta aatcccctta
     2821 tcaatttacg gatgagataa gtcaagctcc tgtattgtat ggctatgccc ggaatactct
     2881 tgaaaacagg ttaaaagcta aatgttgtga attatgtggg acgtctgatg aaaatacttc
     2941 ctatgaaatt caccatgtca ataaggtcaa aaatcttaaa ggcaaagaaa aatgggaaat
     3001 ggcaatgata gcgaaacaac gtaaaactct tgttgtatgc tttcattgtc atcgtcacgt
     3061 gattcataaa cacaagtgaa tgtcgagcac ccgttctcgg agcactgtcc gaccgctttg
     3121 gccgccgccc agtcctgctc gcttcgctac ttggagccac tatcgactac gcgatcatgg
     3181 cgaccacacc cgtcctgtgg atcgccaagc cgccgatggt agtgtggggt ctccccatgc
     3241 gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     3301 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag
     3361 cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa
     3421 ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt gcgtttctac
     3481 aaactcttcc tgtcgtcata tctacaagcc atccccccac agatacgggc gcgccgccat
     3541 tatttttttg aacaattgac aattcatttc ttatttttta ttaagtgata gtcaaaaggc
     3601 ataacagtgc tgaatagaaa gaaatttaca gaaaagaaaa ttatagaatt tagtatgatt
     3661 aattatactc atttatgaat gtttaattga atacaaaaaa aaatacttgt tatgtattca
     3721 attacgggtt aaaatataga caagttgaaa aatttaataa aaaaataagt cctcagctct
     3781 tatatattaa gctaccaact tagtatataa gccaaaactt aaatgtgcta ccaacacatc
     3841 aagccgttag agaactctat ctatagcaat atttcaaatg taccgacata caagagaaac
     3901 attaactata tatattcaat ttatgagatt atcttaacag atataaatgt aaattgcaat
     3961 aagtaagatt tagaagttta tagcctttgt gtattggaag cagtacgcaa aggctttttt
     4021 atttgataaa aattagaagt atatttattt tttcataatt aatttatgaa aatgaaaggg
     4081 ggtgagcaaa gtgacagagg aaagcagtat cttatcaaat aacaaggtat tagcaatatc
     4141 attattgact ttagcagtaa acattatgac ttttatagtg cttgtagcta agtagtacga
     4201 aagggggagc tttaaaaagc tccttggaat acatagaatt cataaattaa tttatgaaaa
     4261 gaagggcgta tatgaaaact tgtaaaaatt gcaaagagtt tattaaagat actgaaatat
     4321 gcaaaataca ttcgttgatg attcatgata aaacagtagc aacctattgc agtaaataca
     4381 atgagtcaag atgtttacat aaagggaaag tccaatgtat taattgttca aagatgaacc
     4441 gatatggatg gtgtgccata aaaatgagat gttttacaga ggaagaacag aaaaaagaac
     4501 gtacatgcat taaatattat gcaaggagct ttaaaaaagc tcatgtaaag aagagtaaaa
     4561 agaaaaaata atttatttat taatttaata ttgagagtgc cgacacagta tgcactaaaa
     4621 aatatatctg tggtgtagtg agccgataca aaaggatagt cactcgcatt ttcataatac
     4681 atcttatgtt atgattatgt gtcggtggga cttcacgacg aaaacccaca ataaaaaaag
     4741 agttcggggt agggttaagc atagttgagg caactaaaca atcaagctag gatatgcagt
     4801 agcagaccgt aaggtcgttg tttaggtgtg ttgtaataca tacgctatta agatgtaaaa
     4861 atacggatac caatgaaggg aaaagtataa tttttggatg tagtttgttt gttcatctat
     4921 gggcaaacta cgtccaaagc cgtttccaaa tctgctaaaa agtatatcct ttctaaaatc
     4981 aaagtcaagt atgaaatcat aaataaagtt taattttgaa gttattatga tattatgttt
     5041 ttctattaaa ataaattaag tatatagaat agtttaataa tagtatatac ttaatgtgat
     5101 aagtgtctga cagtgtcaca gaaaggatga ttgttatgga ttataagcgg ccggccgaag
     5161 caaacttaag agtgtgttga tagtgcagta tcttaaaatt ttgtataata ggaattgaag
     5221 ttaaattaga tgctaaaaat ttgtaattaa gaaggagtga ttacatgaac aaaaatataa
     5281 aatattctca aaacttttta acgagtgaaa aagtactcaa ccaaataata aaacaattga
     5341 atttaaaaga aaccgatacc gtttacgaaa ttggaacagg taaagggcat ttaacgacga
     5401 aactggctaa aataagtaaa caggtaacgt ctattgaatt agacagtcat ctattcaact
     5461 tatcgtcaga aaaattaaaa ctgaatactc gtgtcacttt aattcaccaa gatattctac
     5521 agtttcaatt ccctaacaaa cagaggtata aaattgttgg gagtattcct taccatttaa
     5581 gcacacaaat tattaaaaaa gtggtttttg aaagccatgc gtctgacatc tatctgattg
     5641 ttgaagaagg attctacaag cgtaccttgg atattcaccg aacactaggg ttgctcttgc
     5701 acactcaagt ctcgattcag caattgctta agctgccagc ggaatgcttt catcctaaac
     5761 caaaagtaaa cagtgtctta ataaaactta cccgccatac cacagatgtt ccagataaat
     5821 attggaagct atatacgtac tttgtttcaa aatgggtcaa tcgagaatat cgtcaactgt
     5881 ttactaaaaa tcagtttcat caagcaatga aacacgccaa agtaaacaat ttaagtaccg
     5941 ttacttatga gcaagtattg tctattttta atagttatct attatttaac gggaggaaat
     6001 aattctatga gtcgcttttg taaatttgga aagttacacg ttactaaagg gaatgtgttt
     6061 aaactccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6121 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6181 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6241 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6301 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6361 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6421 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6481 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6541 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6601 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6661 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6721 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6781 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     6841 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     6901 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca gggccccctg cttcggggtc
     6961 attatagcga ttttttcggt atatccatcc tttttcgcac gatatacagg attttgccaa
     7021 agggttcgtg tagactttcc ttggtgtatc caacggcgtc agccgggcag gataggtgaa
     7081 gtaggcccac ccgcgagcgg gtgttccttc ttcactgtcc cttattcgca cctggcggtg
     7141 ctcaacggga atcctgctct gcgaggctgg ccggctaccg ccggcgtaac agatgagggc
     7201 aagcggatgg ctgatgaaac caagccaacc aggaagggca gcccacctat caaggtgtac
     7261 tgccttccag acgaacgaag agcgattgag gaaaaggcgg cggcggccgg catgagcctg
     7321 tcggcctacc tgctggccgt cggccagggc tacaaaatca cgggcgtcgt ggactatgag
     7381 cacgtccgcg agctggcccg catcaatggc gacctgggcc gcctgggcgg cctgctgaaa
     7441 ctctggctca ccgacgaccc gcgcacggcg cggttcggtg atgccacgat cctcgccctg
     7501 ctggcgaaga tcgaagagaa gcaggacgag cttggcaagg tcatgatggg cgtggtccgc
     7561 ccgagggcag agccatgact tttttagccg ctaaaacggc cggggggtgc gcgtgattgc
     7621 caagcacgtc cccatgcgct ccatcaagaa gagcgacttc gcggagctgg tgaagtacat
     7681 caccgacgag caaggcaaga ccgatcgggc c
//
