LOCUS       Exported                7683 bp DNA     circular SYN 01-AUG-2025
DEFINITION  Shuttle expression-secretion vector pP43NMK, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7683)
  AUTHORS   Zhang XZ, Cui ZL, Hong Q, Li SP.
  TITLE     High-level expression and secretion of methyl parathion hydrolase in
            Bacillus subtilis WB800
  JOURNAL   Appl. Environ. Microbiol. 71 (7), 4101-4103 (2005)
  PUBMED    16000826
REFERENCE   2  (bases 1 to 7683)
  AUTHORS   Zhang X-Z., Cui Z-L., Li S-P.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-OCT-2005) Department of Microbiology, College of Life 
            Sciences, Nanjing Agricultural University, MOA Key Lab of 
            Microbiological Engineering of Agricultural Environment, #6 Tongwei 
            Road, Weigang, Nanjing, Jiangsu 210095, P. R. China
REFERENCE   3  (bases 1 to 7683)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7683)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 7683)
  TITLE     Direct Submission
REFERENCE   6  (bases 1 to 7683)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
            Environ. Microbiol."; date: "2005"; volume: "71"; issue: "7"; pages:
            "4101-4103"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (25-OCT-2005) Department of Microbiology, College of Life Sciences,
            Nanjing Agricultural University, MOA Key Lab of Microbiological
            Engineering of Agricultural Environment, #6 Tongwei Road, Weigang,
            Nanjing, Jiangsu 210095, P. R. China"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     SGRef: number: 5; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             814..1818
                     /codon_start=1
                     /gene="repB"
                     /product="RepB replication protein"
                     /label=repB
                     /note="from Enterococcus faecalis plasmid pAM-alpha-1"
                     /db_xref="GI:22652809"
                     /protein_id="AAN03827.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     gene            1987..2748
                     /gene="km"
                     /label=km
     CDS             1987..2748
                     /codon_start=1
                     /gene="km"
                     /product="Km"
                     /label=km
                     /protein_id="ABD24446.1"
                     /translation="MNGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGRQTD
                     GPYSDIEMMCVMSTEEAEFSHEWTTGEWKVEVNFDSEEILLDYASQVESDWPLTHGQFF
                     SILPIYDSGGYLEKVYQTAKSVEAQTFHDAICALIVEELFEYAGKWRNIRVQGPTTFLP
                     SLTVQVAMAGAMLIGLHHRICYTTSASVLTEAVKQSDLPSGYDHLCQFVMSGQLSDSEK
                     LLESLENFWNGIQEWTERHGYIVDVSKRIPF"
     CDS             2965..3369
                     /codon_start=1
                     /gene="ble"
                     /product="Ble"
                     /label=ble
                     /protein_id="ABD24447.1"
                     /translation="MRMLQSIPALPVGDIKKSIGFYCDKLGFTLVHHEDGFAVLMCNEV
                     RIHLWEASDEGWRSRSNDSPVCTGAESFIAGTASCRIEVEGIDELYQHIKPLGILHPNT
                     SLKDQWWDERDFAVIDPDNNLISFFQQIKS"
     promoter        3589..3808
                     /label=Hpa II promoter
                     /note="The HpaII promoter was a strong promoter in 
                     Gram-negative bacteria and  B. subtilis."
     promoter        4366..4421
                     /label=P43
     regulatory      4370..4387
                     /note="-35 and -10 signal of promoter controlled by sigma
                     B"
                     /regulatory_class="minus_35_signal"
     regulatory      4389..4394
                     /note="-35 and -10 signal of promoter controlled by sigma
                     A"
                     /regulatory_class="minus_35_signal"
     regulatory      4403..4412
                     /note="-35 and -10 signal of promoter controlled by sigma
                     B"
                     /regulatory_class="minus_10_signal"
     regulatory      4412..4417
                     /note="-35 and -10 signal of promoter controlled by sigma
                     A"
                     /regulatory_class="minus_10_signal"
     regulatory      4444..4453
                     /gene="mpd"
                     /regulatory_class="ribosome_binding_site"
     CDS             4463..5437
                     /codon_start=1
                     /gene="mpd"
                     /product="MPH"
                     /label=mpd
                     /note="methyl parathion hydrolase; includes NprB signal
                     peptide"
                     /protein_id="ABD24448.1"
                     /translation="MRNSTKTSLLLAGLCTAAQMVFVTHASAAAPQVRTSAPGYYRMLL
                     GDFEITALSDGTVALPVDKRLNQPAPKTQSALAKSFQKAPLETSVTGYLVNTGSKLVLV
                     DTGAAGLFGPTLGRLAANLKAAGYQPEQVDEIYITHMHPDHVGGLMVGEQLAFPNAVVR
                     ADQKEADFWLSQTNLDKAPDDESKGFFKGAMASLNPYVKAGKFKPFSGNTDLVPGIKAL
                     ASHGHTPGHTTYVVESQGQKLALLGDLILVAAVQFDDPSVTNQLDIDGKSAAVERKKAF
                     ADAAKGGYLIAASHLPFPGIGHIRAEGKGYRFVPVNYSVVNPK"
     primer_bind     complement(5462..5478)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5486..5502)
                     /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(5510..5540)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5555..5576)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5864..6452)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6623..7483)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7484..7588)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt ccttaaggaa cgtacagacg
      421 gcttaaaagc ctttaaaaac gtttttaagg ggtttgtaga caaggtaaag gataaaacag
      481 cacaattcca agaaaaacac gatttagaac ctaaaaagaa cgaatttgaa ctaactcata
      541 accgagaggt aaaaaaagaa cgaagtcgag atcagggaat gagtttataa aataaaaaaa
      601 gcacctgaaa aggtgtcttt ttttgatggt tttgaacttg ttctttctta tcttgataca
      661 tatagaaata acgtcatttt tattttagtt gctgaaaggt gcgttgaagt gttggtatgt
      721 atgtgtttta aagtattgaa aacccttaaa attggttgca cagaaaaacc ccatctgtta
      781 aagttataag tgactaaaca aataactaaa tagatggggg tttcttttaa tattatgtgt
      841 cctaatagta gcatttattc agatgaaaaa tcaagggttt tagtggacaa gacaaaaagt
      901 ggaaaagtga gaccatggag agaaaagaaa atcgctaatg ttgattactt tgaacttctg
      961 catattcttg aatttaaaaa ggctgaaaga gtaaaagatt gtgctgaaat attagagtat
     1021 aaacaaaatc gtgaaacagg cgaaagaaag ttgtatcgag tgtggttttg taaatccagg
     1081 ctttgtccaa tgtgcaactg gaggagagca atgaaacatg gcattcagtc acaaaaggtt
     1141 gttgctgaag ttattaaaca aaagccaaca gttcgttggt tgtttctcac attaacagtt
     1201 aaaaatgttt atgatggcga agaattaaat aagagtttgt cagatatggc tcaaggattt
     1261 cgccgaatga tgcaatataa aaaaattaat aaaaatcttg ttggttttat gcgtgcaacg
     1321 gaagtgacaa taaataataa agataattct tataatcagc acatgcatgt attggtatgt
     1381 gtggaaccaa cttattttaa gaatacagaa aactacgtga atcaaaaaca atggattcaa
     1441 ttttggaaaa aggcaatgaa attagactat gatccaaatg taaaagttca aatgattcga
     1501 ccgaaaaata aatataaatc ggatatacaa tcggcaattg acgaaactgc aaaatatcct
     1561 gtaaaggata cggattttat gaccgatgat gaagaaaaga atttgaaacg tttgtctgat
     1621 ttggaggaag gtttacaccg taaaaggtta atctcctatg gtggtttgtt aaaagaaata
     1681 cataaaaaat taaaccttga tgacacagaa gaaggcgatt tgattcatac agatgatgac
     1741 gaaaaagccg atgaagatgg attttctatt attgcaatgt ggaattggga acggaaaaat
     1801 tattttatta aagagtagtt caacaaacgg gccagtttgt tgaagattag atgctataat
     1861 tgttattaaa aggattgaag gatgcttagg aagacgagtt attaatagct gaataagaac
     1921 ggtgctctcc aaatattctt atttagaaaa gcaaatctaa aattatctga aaagggaatg
     1981 agaatagtga atggaccaat aataatgact agagaagaaa gaatgaagat tgttcatgaa
     2041 attaaggaac gaatattgga taaatatggg gatgatgtta aggctattgg tgtttatggc
     2101 tctcttggtc gtcagactga tgggccctat tcggatattg agatgatgtg tgtcatgtca
     2161 acagaggaag cagagttcag ccatgaatgg acaaccggtg agtggaaggt ggaagtgaat
     2221 tttgatagcg aagagattct actagattat gcatctcagg tggaatcaga ttggccgctt
     2281 acacatggtc aatttttctc tattttgccg atttatgatt caggtggata cttagagaaa
     2341 gtgtatcaaa ctgctaaatc ggtagaagcc caaacgttcc acgatgcgat ttgtgccctt
     2401 atcgtagaag agctgtttga atatgcaggc aaatggcgta atattcgtgt gcaaggaccg
     2461 acaacatttc taccatcctt gactgtacag gtagcaatgg caggtgccat gttgattggt
     2521 ctgcatcatc gcatctgtta tacgacgagc gcttcggtct taactgaagc agttaagcaa
     2581 tcagatcttc cttcaggtta tgaccatctg tgccagttcg taatgtctgg tcaactttcc
     2641 gactctgaga aacttctgga atcgctagag aatttctgga atgggattca ggagtggaca
     2701 gaacgacacg gatatatagt ggatgtgtca aaacgcatac cattttgaac gatgacctct
     2761 aataattgtt aatcatgttg gttacgtatt tattaacttc tcctagtatt agtaattatc
     2821 atggctgtca tggcgcatta acggaataaa gggtgtgctt aaatcgggcc attttgcgta
     2881 ataagaaaaa ggattaatta tgagcgaatt gaattaataa taaggtaata gatttacatt
     2941 agaaaatgaa aggggatttt atgcgtgaga atgttacagt ctatcccggc attgccagtc
     3001 ggggatatta aaaagagtat aggtttttat tgcgataaac taggtttcac tttggttcac
     3061 catgaagatg gattcgcagt tctaatgtgt aatgaggttc ggattcatct atgggaggca
     3121 agtgatgaag gctggcgctc tcgtagtaat gattcaccgg tttgtacagg tgcggagtcg
     3181 tttattgctg gtactgctag ttgccgcatt gaagtagagg gaattgatga attatatcaa
     3241 catattaagc ctttgggcat tttgcacccc aatacatcat taaaagatca gtggtgggat
     3301 gaacgagact ttgcagtaat tgatcccgac aacaatttga ttagcttttt tcaacaaata
     3361 aaaagctaaa atctattatt aatctgttca gcaatcgggc gcgattgctg aataaaagat
     3421 acgagagacc tctcttgtat cttttttatt ttgagtggtt ttgtccgtta cactagaaaa
     3481 ccgaaagaca ataaaaattt tattcttgct gagtctggct ttcggtaagc tagacaaaac
     3541 ggacaaaata aaaattggca agggtttaaa ggtggagatt ttttgagtga tcttctcaaa
     3601 aaatactacc tgtcccttgc tgatttttaa acgagcacga gagcaaaacc cccctttgct
     3661 gaggtggcag agggcaggtt tttttgtttc ttttttctcg taaaaaaaag aaaggtctta
     3721 aaggttttat ggttttggtc ggcactgccg acagcctcgc agagcacaca ctttatgaat
     3781 ataaagtata gtgtgttata ctttacttgg aagtggttgc cggaaagagc gaaaatgcct
     3841 catatttgtg ccacctaaaa aggagcgatt tacatatgag ttatgcagtt tgtagaatgc
     3901 aaaaagtgaa atcagctgga ctaaaaggca tgcaatttca taatcaaaga gagcgaaaaa
     3961 gtagaacgaa tgatgatatt gaccatgagc gaacacgtga aaattatgat ttgaaaaatg
     4021 ataaaaatat tgattacaac gaacgtgtca aagaaattat tgaatcacaa aaaacaggta
     4081 caagaaaaac gaggaaagat gctgttcttg taaatgagtt gctagtaaca tctgaccgag
     4141 atttttttga gcaactggat cctgataggt ggtatgtttt cgcttgaact tttaaataca
     4201 gccattgaac atacggttga tttaataact gacaaacatc accctcttgc taaagcggcc
     4261 aaggacgctg ccgccggggc tgtttgcgtt tttaccgtga tttcgtgtat cattggttta
     4321 cttatttttt tgccaaagct gtaatggctg aaaattctta catttatttt acatttttag
     4381 aaatgggcgt gaaaaaaagc gcgcgattat gtaaaatata aagtgatagc ggtaccatta
     4441 tgggtaagag aggaatgtac gcatgcgcaa ctcgaccaag acatctctat tactggccgg
     4501 cttatgcaca gcggcccaaa tggtttttgt aacacatgcc tcagctgcag caccgcaggt
     4561 gcgcacctcg gcccccggct actaccggat gctgctgggc gacttcgaaa tcaccgcgct
     4621 gtcggacggc acggtggcgc tgccggtcga caagcggctg aaccagccgg ccccgaagac
     4681 gcagagcgcg ctggccaagt ccttccagaa agcgccgctc gaaacctcgg tcaccggtta
     4741 cctcgtcaac accggctcca agctggtgct ggtggacacc ggcgcggccg gcctgttcgg
     4801 ccccaccctg ggccggctgg cggccaacct caaggccgca ggctatcagc ccgagcaggt
     4861 cgacgagatc tacatcaccc acatgcaccc cgaccacgtg ggcggcttga tggtgggtga
     4921 gcaactggcg ttcccgaacg cggtggtgcg tgcggaccag aaagaagccg atttctggct
     4981 cagccagacc aacctcgaca aggccccgga cgacgagagc aaaggcttct tcaaaggcgc
     5041 catggcctcg ctgaacccct atgtgaaggc cggcaagttc aagcctttct cggggaacac
     5101 cgacctggtg cccggcatca aagcgctggc cagccacggc cacaccccgg gccacaccac
     5161 ctacgtggtc gaaagccagg ggcaaaagct cgccctgctc ggcgacctga tactcgtcgc
     5221 cgcggtgcag ttcgacgacc ccagcgtcac gaaccagctc gacatcgacg gcaagtccgc
     5281 cgcggtggag cgcaagaagg ccttcgcgga tgccgccaag ggcggctacc tgatcgcggc
     5341 gagccacctg ccgttccccg gcatcggcca catccgcgcc gaaggcaagg gctaccgttt
     5401 cgtgccggtg aactactcgg tcgtcaaccc caagtgatga tgaaagcttg gcgtaatcat
     5461 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5521 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5581 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5641 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5701 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5761 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5821 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5881 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5941 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6001 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6061 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6121 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6181 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6241 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6301 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6361 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     6421 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6481 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6541 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6601 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6661 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6721 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6781 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6841 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6901 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6961 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7021 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7081 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7141 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7201 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7261 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7321 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7381 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7441 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7501 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7561 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     7621 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     7681 gtc
//
