LOCUS       V005453                 8247 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V005453
VERSION     V005453
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8247)
  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 8247)
  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 8247)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8247)
  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..8247
                     /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    complement(674..1067)
                     /note="downstream sequence of Bacillus subtilis 16S rRNA
                     gene"
     repeat_region   1103..1227
     CDS             complement(1261..2340)
                     /label="lacI"
                     /note="lac repressor"
     CDS             complement(2593..2925)
                     /gene="mazF"
                     /label="Endoribonuclease toxin MazF"
                     /note="Endoribonuclease toxin MazF from Escherichia coli
                     (strain K12). Accession#: P0AE70"
     misc_feature    complement(2942..3230)
                     /label="Pspac promoter functional region"
                     /note="Pspac promoter functional region"
     protein_bind    2949..2965
                     /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             3537..4289
                     /codon_start=1
                     /gene="spc"
                     /product="spectinomycin resistance marker"
                     /label="spc"
                     /protein_id="ABC88419.1"
                     /translation="MNTYEQINKVKKILRKHLKNNLIGTYMFGSGVESGLKPNSDLDFL
                     VVVSEPLTDQSKEILIQKIRPISKKIGDKSNLRYIELTIIIQQEMVPWNHPPKQEFIYG
                     EWLQELYEQGYIPQKELNSDLTIMLYQAKRKNKRIYGNYDLEELLPDIPFSDVRRAIMD
                     SSEELIDNYQDDETNSILTLCRMILTMDTGKIIPKDIAGNAVAESSPLEHRERILLAVR
                     SYLGENIEWTNENVNLTINYLNNRLKKL"
     gene            3537..4289
                     /gene="spc"
                     /label="spc"
     repeat_region   4291..4415
     misc_feature    complement(4538..5537)
                     /note="unstream sequence of Bacillus subtilis 16S rRNA
                     gene"
     misc_feature    5544..5791
                     /note="promoter functional region of Escherichia coli lpp
                     gene"
     CDS             5792..6037
                     /gene="mazE"
                     /label="Antitoxin MazE"
                     /note="Antitoxin MazE from Escherichia coli (strain K12).
                     Accession#: P0AE72"
     promoter        complement(6059..6077)
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6098..6114)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(6122..6138)
                     /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(6146..6176)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6191..6212)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6500..7088)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7262..8119)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(8120..8224)
                     /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 gagccgggaa cgtattcacc gcggcatgct gatccgcgat tactagcgat
      721 tccagcttca cgcagtcgag ttgcagactg cgatccgaac tgagaacaga tttgtgggat
      781 tggcttaacc tcgcggttcg ctgccctttg ttctgtccat tgtagcacgt gtgtagccca
      841 ggtcataagg ggcatgatga tttgacgtca tccccacctt cctccggttt gtcaccggca
      901 gtcaccttag agtgcccaac tgaatgctgg caactaagat caagggttgc gctcgttgcg
      961 gcacttaacc caacatctca cgacacgagc tgacgacaac catgcaccac ctgtcactct
     1021 gcccccgaag gggacgtcct atctctagga ttgtcagagg atgtcaaatc gatccgcggc
     1081 tagccatggt cgacgcggcc gcaaaattga aaaaatggtg gaaacacttt tttcaatttt
     1141 tttgttttat tatttaatat ttgggaaata ttcattctaa ttggtaatca gattttagaa
     1201 aacaataaac ccttgcatag ggggatcgga tcctaactca cattaattgc gttgcgctca
     1261 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     1321 gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac
     1381 gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac
     1441 gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttgacggcg ggatataaca
     1501 tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc
     1561 ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc
     1621 agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact
     1681 ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca
     1741 gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg
     1801 ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa
     1861 aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt
     1921 gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc
     1981 actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg
     2041 ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc
     2101 gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga
     2161 ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc
     2221 cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga
     2281 aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcat
     2341 caaaatcgtc tccctccgtt tgaatatttg attgatcgta accagatgaa gcactctttc
     2401 cactatccct acagtgttat ggcttgaaca atcacgaaac aataattggt acgtacgatc
     2461 tttcagccga ctcaaacatc aaatcttaca aatgtagtct ttgaaagtat tacatatgta
     2521 agatttaaat gcaaccgttt tttcggaagg aaatgatgac ctcgtttcca ccggaattag
     2581 cttgcatgcc tacccaatca gtacgttaat tttggcttta atgagttgta attcctctgg
     2641 ggcaactgtt cctttcttcg ttgctcctct tgcccgccag gcgatacttt ttacctgatc
     2701 agctaacgct acgccatcac gttcctgacc ggataaaaca acttcgaacg gatatccttt
     2761 tgattgcgtt gtacaaggaa cacacagaca catacctgtt ttgttgttgt acatgaaagg
     2821 actcaggaca acagctggac gatgtccagc ttgctcgcta ccttttgtcg ggtcaaaatc
     2881 aacccaaatc agatcgccca tatcgggtac gtatcggctt accatacctg cttcctcctt
     2941 aagcttaatt gttatccgct cacaattcca cacattatgc cacaccttgt agataaagtc
     3001 aacaactttt tgcaaaattt ttcaggaatt ttagcagagg ttgttctgga tgtagaacaa
     3061 aacatctttc cgctcttgtg ctgttaggat atctttcttg gaagctaggt aggcaagggc
     3121 tacctcaaat aaatcttctt cagggtgcgc tatttttaag gtgcctagtg aggtcttgac
     3181 cacttcaccc ataatttcag tgccgaatag tctggactgg gctgtgtaga attcctgcag
     3241 ccctggcgaa tggcgatttt cgttcgtgaa tacatgttat aataactata actaataacg
     3301 taacgtgact ggcaagagat atttttaaaa caatgaatag gtttacactt actttagttt
     3361 tatggaaatg aaagatcata tcatatataa tctagaataa aattaactaa aataattatt
     3421 atctagataa aaaatttaga agccaatgaa atctataaat aaactaaatt aagtttattt
     3481 aattaacaac tatggatata aaataggtac taatcaaaat agtgaggagg atatatttga
     3541 atacatacga acaaattaat aaagtgaaaa aaatacttcg gaaacattta aaaaataacc
     3601 ttattggtac ttacatgttt ggatcaggag ttgagagtgg actaaaacca aatagtgatc
     3661 ttgacttttt agtcgtcgta tctgaaccat tgacagatca aagtaaagaa atacttatac
     3721 aaaaaattag acctatttca aaaaaaatag gagataaaag caacttacga tatattgaat
     3781 taacaattat tattcagcaa gaaatggtac cgtggaatca tcctcccaaa caagaattta
     3841 tttatggaga atggttacaa gagctttatg aacaaggata cattcctcag aaggaattaa
     3901 attcagattt aaccataatg ctttaccaag caaaacgaaa aaataaaaga atatacggaa
     3961 attatgactt agaggaatta ctacctgata ttccattttc tgatgtgaga agagccatta
     4021 tggattcgtc agaggaatta atagataatt atcaggatga tgaaaccaac tctatattaa
     4081 ctttatgccg tatgatttta actatggaca cgggtaaaat cataccaaaa gatattgcgg
     4141 gaaatgcagt ggctgaatct tctccattag aacataggga gagaattttg ttagcagttc
     4201 gtagttatct tggagagaat attgaatgga ctaatgaaaa tgtaaattta actataaact
     4261 atttaaataa cagattaaaa aaattataaa aaaattgaaa aaatggtgga aacacttttt
     4321 tcaatttttt tgttttatta tttaatattt gggaaatatt cattctaatt ggtaatcaga
     4381 ttttagaaaa caataaaccc ttgcataggg ggatctcgac atggatgagc gatgatgata
     4441 tccgtttagg ctgggcggtg atagcttctc gttcaggcag tacgcctctt ttcttttcca
     4501 gacctgaggg aggcggaaat ggtgtgaggt tcccggggac ctggtaaggt tcttcgcgtt
     4561 gcttcgaatt aaaccacatg ctccaccgct tgtgcgggcc cccgtcaatt cctttgagtt
     4621 tcagtcttgc gaccgtactc cccaggcgga gtgcttaatg cgttagctgc agcactaagg
     4681 ggcggaaacc ccctaacact tagcactcat cgtttacggc gtggactacc agggtatcta
     4741 atcctgttcg ctccccacgc tttcgctcct cagcgtcagt tacagaccag agagtcgcct
     4801 tcgccactgg tgttcctcca catctctacg catttcaccg ctacacgtgg aattccactc
     4861 tcctcttctg cactcaagtt ccccagtttc caatgaccct ccccggttga gccgggggct
     4921 ttcacatcag acttaagaaa ccgcctgcga gccctttacg cccaataatt ccggacaacg
     4981 cttgccacct acgtattacc gcggctgctg gcacgtagtt agccgtggct ttctggttag
     5041 gtaccgtcaa ggtaccgccc tattcgaacg gtacttgttc ttccctaaca acagagcttt
     5101 acgatccgaa aaccttcatc actcacgcgg cgttgctccg tcagactttc gtccattgcg
     5161 gaagattccc tactgctgcc tcccgtagga gtctgggccg tgtctcagtc ccagtgtggc
     5221 cgatcaccct ctcaggtcgg ctacgcatcg ttgccttggt gagccgttac ctcaccaact
     5281 agctaatgcg ccgcggggcc atctgtaatt ggtaagcgaa gccacctttt atgtttgaac
     5341 catgcggttc aaacaaccat ccggtattag ccccggtttc ccggagttat cccagtctta
     5401 caggcaggtt acccacgtgt tactcacccg tccgccgcta acatcaggga gcaagctccc
     5461 atctgtccgc tcgacttgca tgtattaggc acgccgccag cgttcgtcct gagccaggat
     5521 caaactctcc gataaatact agtcgataac cagaagcaat aaaaaatcaa atcggatttc
     5581 actatataat ctcactttat ctaagatgaa tccgatggaa gcatcctgtt ttctctcaat
     5641 ttttttatct aaaacccagc gttcgatgct tctttgagcg aacgatcaaa aataagtgcc
     5701 ttcccatcaa aaaaatattc tcaacataaa aaactttgtg taatacttgt aacgctacat
     5761 ggagattaac tcaatctaga gggtattaat aatgatccac agtagcgtaa agcgttgggg
     5821 aaattcaccg gcggtgcgga tcccggctac gttaatgcag gcgctcaatc tgaatattga
     5881 tgatgaagtg aagattgacc tggtggatgg caaattaatt attgagccag tgcgtaaaga
     5941 gcccgtattt acgcttgctg aactggtcaa cgacatcacg ccggaaaacc tccacgagaa
     6001 tatcgactgg ggagagccga aagataagga agtctggtaa gagctccagc ttttgttccc
     6061 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     6121 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     6181 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     6241 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     6301 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6361 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6421 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6481 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6541 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6601 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6661 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6721 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6781 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6841 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6901 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     6961 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     7021 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     7081 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     7141 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     7201 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7261 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7321 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7381 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7441 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7501 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7561 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7621 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7681 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7741 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7801 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7861 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7921 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7981 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     8041 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     8101 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     8161 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     8221 cgcgcacatt tccccgaaaa gtgccac
//
