LOCUS       V005452                 8965 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V005452
VERSION     V005452
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8965)
  AUTHORS   Zhang XZ, Yan X, Cui ZL, Hong Q, Li SP.
  TITLE     mazF, a novel counter-selectable marker for unmarked chromosomal
            manipulation in Bacillus subtilis
  JOURNAL   Nucleic Acids Res. 34 (9), E71 (2006)
   PUBMED   16714443
REFERENCE   2  (bases 1 to 8965)
  AUTHORS   Zhang X-Z., Yan X, Li S-P.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-MAR-2006) Department of Microbiology, College of Life
            Sciences, Nanjing Agricultural University, Key Laboratory for
            Microbiological Engineering of Agricultural Environment, Ministry of
            Agriculture, 6 Tongwei Road, Nanjing, Jiangsu 210095, China
REFERENCE   3  (bases 1 to 8965)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8965)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
            Acids Res."; date: "2006"; volume: "34"; issue: "9"; pages: "E71"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (21-MAR-2006) Department of Microbiology, College of Life Sciences,
            Nanjing Agricultural University, Key Laboratory for Microbiological
            Engineering of Agricultural Environment, Ministry of Agriculture, 6
            Tongwei Road, Nanjing, Jiangsu 210095, China"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8965
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      4..459
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        626..644
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    692..1697
                     /note="bpr-front; upstream of Bacillus subtilis bpr gene;
                     homologous arm for recombination"
     repeat_region   1728..1852
                     /label="DR sequence"
                     /note="DR sequence"
     CDS             complement(1903..2982)
                     /label="lacI"
                     /note="lac repressor"
     CDS             complement(3235..3567)
                     /gene="mazF"
                     /label="Endoribonuclease toxin MazF"
                     /note="Endoribonuclease toxin MazF from Escherichia coli
                     (strain K12). Accession#: P0AE70"
     protein_bind    3591..3607
                     /label="lac operator"
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             4179..4931
                     /codon_start=1
                     /gene="spc"
                     /product="Spc"
                     /label="spc"
                     /note="spectinomycin adenyltransferase; selectable either
                     in Escherichia coli or Bacillus subtilis"
                     /protein_id="ABF31010.1"
                     /translation="MNTYEQINKVKKILRKHLKNNLIGTYMFGSGVESGLKPNSDLDFL
                     VVVSEPLTDQSKEILIQKIRPISKKIGDKSNLRYIELTIIIQQEMVPWNHPPKQEFIYG
                     EWLQELYEQGYIPQKELNSDLTIMLYQAKRKNKRIYGNYDLEELLPDIPFSDVRRAIMD
                     SSEELIDNYQDDETNSILTLCRMILTMDTGKIIPKDIAGNAVAESSPLEHRERILLAVR
                     SYLGENIEWTNENVNLTINYLNNRLKKL"
     gene            4179..4931
                     /gene="spc"
                     /label="spc"
     repeat_region   4933..5057
                     /label="DR sequence"
                     /note="DR sequence"
     misc_feature    5197..6235
                     /note="bpr-back; downstream of Bacillus subtilis bpr gene;
                     homologous arm for recombination"
     CDS             6510..6755
                     /gene="mazE"
                     /label="Antitoxin MazE"
                     /note="Antitoxin MazE from Escherichia coli (strain K12).
                     Accession#: P0AE72"
     promoter        complement(6777..6795)
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6816..6832)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(6840..6856)
                     /label="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(6864..6894)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6909..6930)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7218..7806)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7980..8837)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(8838..8942)
                     /label="AmpR promoter"
ORIGIN
        1 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
       61 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      121 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      181 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      241 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      301 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
      361 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
      421 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
      661 gccccccctc gagttaacct gcaggcgcgc ctttgatggg tatcggtatt gctactgggg
      721 aaaatcgcgc ggcagaggca gcaaaaaaag caatttccag cccgcttctt gaagcggcca
      781 ttgacggtgc gcaaggcgtc ctcatgaaca tcactggagg aacaaacctc agcctatatg
      841 aggttcagga agcagcagac attgtcgctt cggcgtctga tcaagacgta aacatgattt
      901 tcggttctgt tattaatgaa aatctaaaag atgagattgt ggtgacagtg attgcaaccg
      961 gctttatcga acaagagaag gacgtgacga agcctcagcg cccaagctta aatcaaagca
     1021 tcaaaacaca caatcaaagt gttccgaagc gtgacgcaaa acgtgaggaa cctcagcagc
     1081 agaacacagt aagccgtcat acttcacagc cggctgatga tacgcttgac atcccgacat
     1141 tcttaagaaa ccgtaataaa cgcggctaat gtaaaggaca aaatcgtttt cgattttgtc
     1201 ttttttgttt ttctcttcac acttccttct tataaagtct ttttccctat tgctttcctt
     1261 cgcttagtaa caaaacagat aattagaccc atttattttt gtgacatttt tatcattttc
     1321 atatatatgg aaattgaatg acatgaaacg acaatatctg taattcagat tgtctacagt
     1381 taatatacag cgatgttctg acaaaccatt cattattaaa aggagggacg acactttttt
     1441 taaaaagcat gttgaaaaag ggggatgaaa atgaggaaaa aaacgaaaaa cagactcatc
     1501 agctctgttt taagtacagt tgtcatcagt tcactgctgt ttccgggagc agccggggca
     1561 agcagtaaag tcacctcacc ttctgttaaa aaggagcttc aatctgcgga atccattcaa
     1621 aacaagattt cgagttcatt aaagaaaagc tttaaaaaga aagaaaaaac gacttttctg
     1681 attaaattta aagatctatc gatgctagcc atggtcgacg cggccgcaaa attgaaaaaa
     1741 tggtggaaac acttttttca atttttttgt tttattattt aatatttggg aaatattcat
     1801 tctaattggt aatcagattt tagaaaacaa taaacccttg cataggggga tctccggagt
     1861 ttaaaccgcg gatcctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     1921 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     1981 cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc
     2041 cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag
     2101 gcgaaaatcc tgtttgatgg tggttgacgg cgggatataa catgagctgt cttcggtatc
     2161 gtcgtatccc actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg
     2221 cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc
     2281 attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc
     2341 cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg
     2401 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac
     2461 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg
     2521 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc
     2581 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag
     2641 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac
     2701 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc
     2761 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg
     2821 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg
     2881 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac
     2941 accggcatac tctgcgacat cgtataacgt tactggtttc atcaaaatcg tctccctccg
     3001 tttgaatatt tgattgatcg taaccagatg aagcactctt tccactatcc ctacagtgtt
     3061 atggcttgaa caatcacgaa acaataattg gtacgtacga tctttcagcc gactcaaaca
     3121 tcaaatctta caaatgtagt ctttgaaagt attacatatg taagatttaa atgcaaccgt
     3181 tttttcggaa ggaaatgatg acctcgtttc caccggaatt agcttgcatg cctacccaat
     3241 cagtacgtta attttggctt taatgagttg taattcctct ggggcaactg ttcctttctt
     3301 cgttgctcct cttgcccgcc aggcgatact ttttacctga tcagctaacg ctacgccatc
     3361 acgttcctga ccggataaaa caacttcgaa cggatatcct tttgattgcg ttgtacaagg
     3421 aacacacaga cacatacctg ttttgttgtt gtacatgaaa ggactcagga caacagctgg
     3481 acgatgtcca gcttgctcgc taccttttgt cgggtcaaaa tcaacccaaa tcagatcgcc
     3541 catatcgggt acgtatcggc ttaccatacc tgcttcctcc ttaagcttaa ttgttatccg
     3601 ctcacaattc cacacattat gccacacctt gtagataaag tcaacaactt tttgcaaaat
     3661 ttttcaggaa ttttagcaga ggttgttctg gatgtagaac aaaacatctt tccgctcttg
     3721 tgctgttagg atatctttct tggaagctag gtaggcaagg gctacctcaa ataaatcttc
     3781 ttcagggtgc gctattttta aggtgcctag tgaggtcttg accacttcac ccataatttc
     3841 agtgccgaat agtctggact gggctgtgta gaattcctgc agccctggcg aatggcgatt
     3901 ttcgttcgtg aatacatgtt ataataacta taactaataa cgtaacgtga ctggcaagag
     3961 atatttttaa aacaatgaat aggtttacac ttactttagt tttatggaaa tgaaagatca
     4021 tatcatatat aatctagaat aaaattaact aaaataatta ttatctagat aaaaaattta
     4081 gaagccaatg aaatctataa ataaactaaa ttaagtttat ttaattaaca actatggata
     4141 taaaataggt actaatcaaa atagtgagga ggatatattt gaatacatac gaacaaatta
     4201 ataaagtgaa aaaaatactt cggaaacatt taaaaaataa ccttattggt acttacatgt
     4261 ttggatcagg agttgagagt ggactaaaac caaatagtga tcttgacttt ttagtcgtcg
     4321 tatctgaacc attgacagat caaagtaaag aaatacttat acaaaaaatt agacctattt
     4381 caaaaaaaat aggagataaa agcaacttac gatatattga attaacaatt attattcagc
     4441 aagaaatggt accgtggaat catcctccca aacaagaatt tatttatgga gaatggttac
     4501 aagagcttta tgaacaagga tacattcctc agaaggaatt aaattcagat ttaaccataa
     4561 tgctttacca agcaaaacga aaaaataaaa gaatatacgg aaattatgac ttagaggaat
     4621 tactacctga tattccattt tctgatgtga gaagagccat tatggattcg tcagaggaat
     4681 taatagataa ttatcaggat gatgaaacca actctatatt aactttatgc cgtatgattt
     4741 taactatgga cacgggtaaa atcataccaa aagatattgc gggaaatgca gtggctgaat
     4801 cttctccatt agaacatagg gagagaattt tgttagcagt tcgtagttat cttggagaga
     4861 atattgaatg gactaatgaa aatgtaaatt taactataaa ctatttaaat aacagattaa
     4921 aaaaattata aaaaaattga aaaaatggtg gaaacacttt tttcaatttt tttgttttat
     4981 tatttaatat ttgggaaata ttcattctaa ttggtaatca gattttagaa aacaataaac
     5041 ccttgcatag ggggatctcg acatggatga gcgatgatga tatccgttta ggctgggcgg
     5101 tgatagcttc tcgttcaggc agtacgcctc ttttcttttc cagacctgag ggaggcggaa
     5161 atggtgtgag gttcccgggc cggccaattg atgcatcagt tcaaaaggga ttatggtcga
     5221 taaagatttt tatctcgtat atatccagtc gaaacctgat ccgtattcac ctggattggc
     5281 aatggatgaa accggccaga attccggccg caactggcag tatatagatg gaaagtggca
     5341 gccaggtgac aaagcggatg gcaactatat gattcgcgca ttagttgatt atgaagctgc
     5401 tgtacctgag attacttcac cgacagacaa atcatacaca aataaggata gcgtcactgt
     5461 aaaaggaaac gcttctcctg gcacaacggt acacatttat aatggagaga aagaagcagg
     5521 agaaacgaaa gctgctgcgg atggcacgtt ccatgcaggc atcatactca acaagggtga
     5581 aaatgagctg acggcaactg catcaactga caacggaaca acagatgcct ccagcccaat
     5641 cacggtcacg cttgatcaag aaaagcctga attaacactg gacaatccaa aggatggcgg
     5701 gaaaacaaat aaagaaacgc tgactgtcaa aggggctgta tccgatgaca atctgaaaga
     5761 cgtcaaggtg aatggcaaaa aagcaacagt agctgatggt tcatactcag cccgtattct
     5821 tttggaaaat ggaagaaatg aaatcaaggt aattgctaca gacttggcag gcaacaaaac
     5881 gacgaaaaag acagtcattg atgtgaactt tgacaagcct gtcatttccg gcttaattcc
     5941 gggagaggat aaaaacttaa aagccggtga atctgtgaaa atcgctttct caagcgctga
     6001 ggatttagat gcaacgttta ccattcgtat gccgctgacc aatgcaagag cgagtgtgca
     6061 aaatgccacc gaactcccgt taagagaaat ctctccgggg agatatgaag gctattggac
     6121 tgccacttct tctattaaag caaaaggagc aaaagtagaa gtgatcgtcc gagatgatta
     6181 tggaaatgaa acaagaaaaa ctgcgaatgg aaaacttaat atgaacacag aaaataccgg
     6241 tcctagggcg atcgcactag tcgataacca gaagcaataa aaaatcaaat cggatttcac
     6301 tatataatct cactttatct aagatgaatc cgatggaagc atcctgtttt ctctcaattt
     6361 ttttatctaa aacccagcgt tcgatgcttc tttgagcgaa cgatcaaaaa taagtgcctt
     6421 cccatcaaaa aaatattctc aacataaaaa actttgtgta atacttgtaa cgctacatgg
     6481 agattaactc aatctagagg gtattaataa tgatccacag tagcgtaaag cgttggggaa
     6541 attcaccggc ggtgcggatt ccggctacgt taatgcaggc gctcaatctg aatattgatg
     6601 atgaagtgaa gattgacctg gtggatggca aattaattat tgagccagtg cgtaaagagc
     6661 ccgtatttac gcttgctgaa ctggtcaacg acatcacgcc ggaaaacctc cacgagaata
     6721 tcgactgggg agagccgaaa gataaggaag tctggtaaga gctccagctt ttgttccctt
     6781 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     6841 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     6901 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     6961 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     7021 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     7081 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     7141 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     7201 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     7261 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     7321 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     7381 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     7441 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     7501 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     7561 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     7621 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     7681 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     7741 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     7801 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7861 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7921 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7981 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     8041 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     8101 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     8161 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     8221 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     8281 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     8341 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     8401 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     8461 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     8521 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     8581 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     8641 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     8701 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     8761 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     8821 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     8881 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     8941 cgcacatttc cccgaaaagt gccac
//
