LOCUS       V008011                 8717 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V008011
VERSION     V008011
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8717)
  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 8717)
  AUTHORS   Zhang X-Z., Li S-P.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-JAN-2006) Department of Microbiology, College of Life
            Sciences, Nanjing Agricultral University, Key Laboratory for
            Microbiological Engineering of Agricultural Environment of Ministry
            of Agriculture, 6 Tongwei Road, Nanjing, Jiangsu 210095, China
REFERENCE   3  (bases 1 to 8717)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8717)
  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
            (10-JAN-2006) Department of Microbiology, College of Life Sciences,
            Nanjing Agricultral University, Key Laboratory for Microbiological
            Engineering of Agricultural Environment of Ministry of Agriculture,
            6 Tongwei Road, Nanjing, Jiangsu 210095, China"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8717
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    115..635
                     /label="unstream sequence of Bacillus subtilis amyE gene"
                     /note="unstream sequence of Bacillus subtilis amyE gene"
     repeat_region   668..792
     CDS             complement(826..1905)
                     /label="lacI"
                     /note="lac repressor"
     CDS             complement(2158..2490)
                     /gene="mazF"
                     /label="Endoribonuclease toxin MazF"
                     /note="Endoribonuclease toxin MazF from Escherichia coli
                     (strain K12). Accession#: P0AE70"
     protein_bind    complement(2514..2530)
                     /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)."
     CDS             3102..3854
                     /codon_start=1
                     /gene="spc"
                     /product="spectinomycin resistance marker"
                     /label="spc"
                     /protein_id="ABC88414.1"
                     /translation="MNTYEQINKVKKILRKHLKNNLIGTYMFGSGVESGLKPNSDLDFL
                     VVVSEPLTDQSKEILIQKIRPISKKIGDKSNLRYIELTIIIQQEMVPWNHPPKQEFIYG
                     EWLQELYEQGYIPQKELNSDLTIMLYQAKRKNKRIYGNYDLEELLPDIPFSDVRRAIMD
                     SSEELIDNYQDDETNSILTLCRMILTMDTGKIIPKDIAGNAVAESSPLEHRERILLAVR
                     SYLGENIEWTNENVNLTINYLNNRLKKL"
     gene            3102..3854
                     /gene="spc"
                     /label="spc"
     repeat_region   3856..3980
     misc_feature    3981..5009
                     /label="downstream sequence of Bacillus subtilis amyE gene"
                     /note="downstream sequence of Bacillus subtilis amyE gene"
     misc_feature    5643..5890
                     /note="promoter functional region of Escherichia coli lpp
                     gene"
     CDS             5891..6136
                     /gene="mazE"
                     /label="Antitoxin MazE"
                     /note="Antitoxin MazE from Escherichia coli (strain K12).
                     Accession#: P0AE72"
     rep_origin      complement(6888..7476)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7650..8507)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(8508..8612)
                     /label="AmpR promoter"
ORIGIN
        1 aacaaaattc tccagtcttc acatcggttt gaaaggagga agcggaagaa tgaagtaaga
       61 gggatttttg actccgaagt aagtcttcaa aaaatcaaat aaggagtgtc aagaatgttt
      121 gcaaaacgat tcaaaacctc tttactgccg ttattcgctg gatttttatt gctgtttcat
      181 ttggttctgg caggaccggc ggctgcgagt gctgaaacgg cgaacaaatc gaatgagctt
      241 acagcaccgt cgatcaaaag cggaaccatt cttcatgcat ggaattggtc gttcaatacg
      301 ttaaaacaca atatgaagga tattcatgat gcaggatata cagccattca gacatctccg
      361 attaaccaag taaaggaagg gaatcaagga gataaaagca tgtcgaactg gtactggctg
      421 tatcagccga catcgtatca aattggcaac cgttacttag gtactgaaca agaatttaaa
      481 gaaatgtgtg cagccgctga agaatatggc ataaaggtca ttgttgacgc ggtcatcaat
      541 cataccacca gtgattatgc cgcgatttcc aatgaggtta agagtattcc aaactggaca
      601 catggaaaca cacaaattaa aaactggtct gatcggatcc gcggctagcc atggtcgacg
      661 cggccgcaaa attgaaaaaa tggtggaaac acttttttca atttttttgt tttattattt
      721 aatatttggg aaatattcat tctaattggt aatcagattt tagaaaacaa taaacccttg
      781 cataggggga tcggatccta actcacatta attgcgttgc gctcactgcc cgctttccag
      841 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
      901 ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt
      961 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
     1021 caggcgaaaa tcctgtttga tggtggttga cggcgggata taacatgagc tgtcttcggt
     1081 atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc
     1141 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc
     1201 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
     1261 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag
     1321 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
     1381 gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat
     1441 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
     1501 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
     1561 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
     1621 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
     1681 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
     1741 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
     1801 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
     1861 gacaccggca tactctgcga catcgtataa cgttactggt ttcatcaaaa tcgtctccct
     1921 ccgtttgaat atttgattga tcgtaaccag atgaagcact ctttccacta tccctacagt
     1981 gttatggctt gaacaatcac gaaacaataa ttggtacgta cgatctttca gccgactcaa
     2041 acatcaaatc ttacaaatgt agtctttgaa agtattacat atgtaagatt taaatgcaac
     2101 cgttttttcg gaaggaaatg atgacctcgt ttccaccgga attagcttgc atgcctaccc
     2161 aatcagtacg ttaattttgg ctttaatgag ttgtaattcc tctggggcaa ctgttccttt
     2221 cttcgttgct cctcttgccc gccaggcgat actttttacc tgatcagcta acgctacgcc
     2281 atcacgttcc tgaccggata aaacaacttc gaacggatat ccttttgatt gcgttgtaca
     2341 aggaacacac agacacatac ctgttttgtt gttgtacatg aaaggactca ggacaacagc
     2401 tggacgatgt ccagcttgct cgctaccttt tgtcgggtca aaatcaaccc aaatcagatc
     2461 gcccatatcg ggtacgtatc ggcttaccat acctgcttcc tccttaagct taattgttat
     2521 ccgctcacaa ttccacacat tatgccacac cttgtagata aagtcaacaa ctttttgcaa
     2581 aatttttcag gaattttagc agaggttgtt ctggatgtag aacaaaacat ctttccgctc
     2641 ttgtgctgtt aggatatctt tcttggaagc taggtaggca agggctacct caaataaatc
     2701 ttcttcaggg tgcgctattt ttaaggtgcc tagtgaggtc ttgaccactt cacccataat
     2761 ttcagtgccg aatagtctgg actgggctgt gtagaattcc tgcagccctg gcgaatggcg
     2821 attttcgttc gtgaatacat gttataataa ctataactaa taacgtaacg tgactggcaa
     2881 gagatatttt taaaacaatg aataggttta cacttacttt agttttatgg aaatgaaaga
     2941 tcatatcata tataatctag aataaaatta actaaaataa ttattatcta gataaaaaat
     3001 ttagaagcca atgaaatcta taaataaact aaattaagtt tatttaatta acaactatgg
     3061 atataaaata ggtactaatc aaaatagtga ggaggatata tttgaataca tacgaacaaa
     3121 ttaataaagt gaaaaaaata cttcggaaac atttaaaaaa taaccttatt ggtacttaca
     3181 tgtttggatc aggagttgag agtggactaa aaccaaatag tgatcttgac tttttagtcg
     3241 tcgtatctga accattgaca gatcaaagta aagaaatact tatacaaaaa attagaccta
     3301 tttcaaaaaa aataggagat aaaagcaact tacgatatat tgaattaaca attattattc
     3361 agcaagaaat ggtaccgtgg aatcatcctc ccaaacaaga atttatttat ggagaatggt
     3421 tacaagagct ttatgaacaa ggatacattc ctcagaagga attaaattca gatttaacca
     3481 taatgcttta ccaagcaaaa cgaaaaaata aaagaatata cggaaattat gacttagagg
     3541 aattactacc tgatattcca ttttctgatg tgagaagagc cattatggat tcgtcagagg
     3601 aattaataga taattatcag gatgatgaaa ccaactctat attaacttta tgccgtatga
     3661 ttttaactat ggacacgggt aaaatcatac caaaagatat tgcgggaaat gcagtggctg
     3721 aatcttctcc attagaacat agggagagaa ttttgttagc agttcgtagt tatcttggag
     3781 agaatattga atggactaat gaaaatgtaa atttaactat aaactattta aataacagat
     3841 taaaaaaatt ataaaaaaat tgaaaaaatg gtggaaacac ttttttcaat ttttttgttt
     3901 tattatttaa tatttgggaa atattcattc taattggtaa tcagatttta gaaaacaata
     3961 aacccttgca tagggggatc tcgacatgga tgagcgatga tgatatccgt ttaggctggg
     4021 cggtgatagc ttctcgttca ggcagtacgc ctcttttctt ttccagacct gagggaggcg
     4081 gaaatggtgt gaggttcccg gggaaaagcc aaataggcga tcgcgggagt gctttatttg
     4141 aagatcaggc tatcactgcg gtcaatagat ttcacaatgt gatggctgga cagcctgagg
     4201 aactctcgaa cccgaatgga aacaaccaga tatttatgaa tcagcgcggc tcacatggcg
     4261 ttgtgctggc aaatgcaggt tcatcctctg tctctatcaa tacggcaaca aaattgcctg
     4321 atggcaggta tgacaataaa gctggagcgg gttcatttca agtgaacgat ggtaaactga
     4381 caggcacgat caatgccagg tctgtagctg tgctttatcc tgatgatatt gcaaaagcgc
     4441 ctcatgtttt ccttgagaat tacaaaacag gtgtaacaca ttctttcaat gatcaactga
     4501 cgattacctt gcgtgcagat gcgaatacaa caaaagccgt ttatcaaatc aataatggac
     4561 cagacgacag gcgtttaagg atggagatca attcacaatc ggaaaaggag atccaatttg
     4621 gcaaaacata caccatcatg ttaaaaggaa cgaacagtga tggtgtaacg aggaccgaga
     4681 aatacagttt tgttaaaaga gatccagcgt cggccaaaac catcggctat caaaatccga
     4741 atcattggag ccaggtaaat gcttatatct ataaacatga tgggagccga gtaattgaat
     4801 tgaccggatc ttggcctgga aaaccaatga ctaaaaatgc agacggaatt tacacgctga
     4861 cgctgcctgc ggacacggat acaaccaacg caaaagtgat ttttaataat ggcagcgccc
     4921 aagtgcccgg tcagaatcag cctggctttg attacgtgct aaatggttta tataatgact
     4981 cgggcttaag cggttctctt ccccattgag ggcaaggcta gacgggactt accgaaagaa
     5041 accatcaatg atggtttctt ttttgttcat aaatcagaca aaacttttct cttgcaaaag
     5101 tttgtgaagt gttgcacaat ataaatgtga aatacttcac aaacaaaaag acatcaaaga
     5161 gaaacatacc ctgcaaggat gctgatattg tctgcatttg cgccggagca aaccaaaaac
     5221 ctggtgagac acgccttgaa ttagtagaaa agaacttgaa gattttcaaa ggcatcgtta
     5281 gtgaagtcat ggcgagcgga tttgacggca ttttcttagt cgcgacgcga ggctggatgg
     5341 ccttccccat tatgattctt ctcgcttccg gcggcatcgg gatgcccgcg ttgcaggcca
     5401 tgctgtccag gcaggtagat gacgaccatc agggacagct tcaaggatcg ctcgcggctc
     5461 ttaccagcct aacttcgatc actggaccgc tgatcgtcac ggcgatttat gccgcctcgg
     5521 cgagcacatg gaacgggttg gcatggattg taggcgccgc cctatacctt gtctgcctcc
     5581 ccgcgttgcg tcgcggtgca tggagccggg ccacctactg aagtggattt ctttaagagc
     5641 tccgataacc agaagcaata aaaaatcaaa tcggatttca ctatataatc tcactttatc
     5701 taagatgaat ccgatggaag catcctgttt tctctcaatt tttttatcta aaacccagcg
     5761 ttcgatgctt ctttgagcga acgatcaaaa ataagtgcct tcccatcaaa aaaatattct
     5821 caacataaaa aactttgtgt aatacttgta acgctacatg gagattaact caatctagag
     5881 ggtattaata atgatccaca gtagcgtaaa gcgttgggga aattcaccgg cggtgcggat
     5941 cccggctacg ttaatgcagg cgctcaatct gaatattgat gatgaagtga agattgacct
     6001 ggtggatggc aaattaatta ttgagccagt gcgtaaagag cccgtattta cgcttgctga
     6061 actggtcaac gacatcacgc cggaaaacct ccacgagaat atcgactggg gagagccgaa
     6121 agataaggaa gtctggtaaa tcgattatgt cttttgcgca gtcggcttaa accagttttc
     6181 gctggtgcga aaaaagagtg tcttgtgaca cctaaattca aaatctatcg gtcagattta
     6241 taccgatttg attttatata ttcttgaata acatacgccg agttatcaca taaaagcggg
     6301 aaccaatcat caaatttaaa cttcattgca taatccatta aactcttaaa ttctacgatt
     6361 ccttgttcat caataaactc aatcatttct ttaattaatt tatatctatc tgttgttgtt
     6421 ttctttaata attcatcaac atctacaccg ccataaacta tcatatcttc tttttgatat
     6481 ttaaatttat taggatctta aggcctaggt ctagagtctt tgttttgacg ccattagcgt
     6541 acgtaacaat cctcgttaaa ggacaaggac ctgagcggaa gtgtatcgta cagtagacgg
     6601 agtatactag tatagtctat agtccgtgga attattatat ttatctccga cgatattctc
     6661 atcagtgaaa tccagctgga gttctttagc aaattttttt attagctgaa cttagtatta
     6721 gtggccatac tcctccaatc caaagctatt tagaaagatt actatatcct caaacaggcg
     6781 gtaaccggcc tcttcatcgg gaatgcgcgc gaccttcagc atcgccggca tgtccccctg
     6841 gcggacggga agtatccagc tcgaggtcgg gccgcgttgc tggcgttttt ccataggctc
     6901 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6961 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     7021 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     7081 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     7141 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     7201 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     7261 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     7321 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7381 gttgatagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7441 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7501 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7561 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7621 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7681 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7741 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7801 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7861 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7921 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca
     7981 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     8041 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     8101 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     8161 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     8221 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg
     8281 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     8341 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     8401 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     8461 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     8521 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     8581 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     8641 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     8701 tttcgtcttc aagaatt
//
