LOCUS       40924_34022        7680 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle expression-secretion vector pP43NMK, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7680)
  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 7680)
  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 7680)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7680)
  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"
FEATURES             Location/Qualifiers
     source          1..7680
                     /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..1815
                     /label=repB
                     /note="RepB replication protein"
     CDS             1987..2748
                     /codon_start=1
                     /gene="km"
                     /product="Km"
                     /label=km
                     /protein_id="ABD24446.1"
                     /translation="MNGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGRQTD
                     GPYSDIEMMCVMSTEEAEFSHEWTTGEWKVEVNFDSEEILLDYASQVESDWPLTHGQFF
                     SILPIYDSGGYLEKVYQTAKSVEAQTFHDAICALIVEELFEYAGKWRNIRVQGPTTFLP
                     SLTVQVAMAGAMLIGLHHRICYTTSASVLTEAVKQSDLPSGYDHLCQFVMSGQLSDSEK
                     LLESLENFWNGIQEWTERHGYIVDVSKRIPF"
     gene            1987..2748
                     /gene="km"
                     /label=km
     CDS             2965..3216
                     /codon_start=1
                     /gene="ble"
                     /product="Ble"
                     /label=ble
                     /protein_id="ABD24447.1"
                     /translation="MRMLQSIPALPVGDIKKSIGFYWDKVGFTLVHHEDGFAVLMCNEV
                     RIHLWEASDEGWRLVVMIHRFVQVRSRLLLVLLVAALK"
     gene            2965..3216
                     /gene="ble"
                     /label=ble
     regulatory      4367..4384
                     /note="-35 and -10 signal of promoter controlled by sigma
                     B"
                     /regulatory_class="minus_35_signal"
     regulatory      4386..4391
                     /note="-35 and -10 signal of promoter controlled by sigma
                     A"
                     /regulatory_class="minus_35_signal"
     regulatory      4400..4409
                     /note="-35 and -10 signal of promoter controlled by sigma
                     B"
                     /regulatory_class="minus_10_signal"
     regulatory      4409..4414
                     /note="-35 and -10 signal of promoter controlled by sigma
                     A"
                     /regulatory_class="minus_10_signal"
     mRNA            4419..5434
                     /gene="mpd"
                     /product="MPH"
                     /label=mpd mRNA
     gene            4419..5434
                     /gene="mpd"
                     /label=mpd
     regulatory      4441..4450
                     /gene="mpd"
                     /regulatory_class="ribosome_binding_site"
     CDS             4460..5434
                     /codon_start=1
                     /gene="mpd"
                     /product="MPH"
                     /label=mpd
                     /note="methyl parathion hydrolase; includes NprB signal
                     peptide"
                     /protein_id="ABD24448.1"
                     /translation="MRNLTKTSLLLAGLCTAAQMVFVTHASAAAPQVRTSAPGYYRMLL
                     GDFEITALSDGTVALPVDKRLNQPAPKTQSALAKSFQKAPLETSVTGYLVNTGSKLVLV
                     DTGAAGLFGPTLGRLAANLKAAGYQPEQVDEIYITHMHPDHVGGLMVGEQLAFPNAVVR
                     ADQKEADFWLSQTNLDKAPDDESKGFFKGAMASLNPYVKAGKFKPFSGNTDLVPGIKAL
                     ASHGHTPGHTTYVVESQGQKLALLGDLILVAAVQFDDPSVTNQLDIDGKSAAVERKKAF
                     ADAAKGGYLIAASHLPFPGIGHIRAEGKGYRFVPVNYSVVNPK"
     sig_peptide     4460..4546
                     /gene="mpd"
                     /note="from NprB"
     primer_bind     complement(5459..5475)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5483..5499)
                     /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(5507..5537)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5552..5573)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5861..6449)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6623..7480)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7481..7585)
                     /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 tgggataaag taggtttcac tttggttcac
     3061 catgaagatg gattcgcagt tctaatgtgt aatgaggttc ggattcatct atgggaggca
     3121 agtgatgaag gctggcgcct cgtagtaatg attcaccggt ttgtacaggt gcggagtcgt
     3181 ttattgctgg tactgctagt tgccgcattg aagtagaggg aattgatgaa ttatatcaac
     3241 atattaagcc tttgggcatt ttgcacccca atacatcatt aaaagatcag tggtgggatg
     3301 aacgagactt tgcagtaatt gatcccgaca acaatttgat tagctttttt caacaaataa
     3361 aaagctaaaa tctattatta atctgttcag caatcgggcg cgattgctga ataaaagata
     3421 cgagagacct ctcttgtatc ttttttattt tgagtggttt tgtccgttac actagaaaac
     3481 cgaaagacaa taaaaatttt attcttgctg agtctggctt tcggtaagct agacaaaacg
     3541 gacaaaataa aaattggcaa gggtttaaag gtggagattt tttgagtgat cttctcaaaa
     3601 aatactacct gtcccttgct gatttttaaa cgagcacgag agcaaaaccc ccctttgctg
     3661 aggtggcaga gggcaggttt ttttgtttct tttttctcgt aaaaaaaaga aaggtcttaa
     3721 aggttttatg gttttggtcg gcactgccgc gcctcgcaga gcacacactt tatgaatata
     3781 aagtatagtg tgttatactt tacttggaag tggttgccgg aaagagcgaa aatgcctcac
     3841 atttgtgcca cctaaaaagg agcgatttac atatgagtta tgcagtttgt agaatgcaaa
     3901 aagtgaaatc agctggacta aaaggcatgc aatttcataa tcaaagagag cgaaaaagta
     3961 gaacgaatga tgatattgac catgagcgaa cacgtgaaaa ttatgatttg aaaaatgata
     4021 aaaatattga ttacaacgaa cgtgtcaaag aaattattga atcacaaaaa acaggtacaa
     4081 gaaaaacgag gaaagatgct gttcttgtaa atgagttgct agtaacatct gaccgagatt
     4141 tttttgagca actggatcct gataggtggt atgttttcgc ttgaactttt aaatacagcc
     4201 attgaacata cggttgattt aataactgac aaacatcacc ctcttgctaa agcggccaag
     4261 gacgctgccg ccggggctgt ttgcgttttt gccgtgattt cgtgtatcat tggtttactt
     4321 atttttttgc caaagctgta atggctgaaa attcttacat ttattttaca tttttagaaa
     4381 tgggcgtgaa aaaaagcgcg cgattatgta aaatataaag tgatagcggt accattatag
     4441 gtaagagagg aatgtacaca tgcgcaactt gaccaagaca tctctattac tggccggctt
     4501 atgcacagcg gcccaaatgg tttttgtaac acatgcctca gctgcagcac cgcaggtgcg
     4561 cacctcggcc cccggctact accggatgct gctgggcgac ttcgaaatca ccgcgctgtc
     4621 ggacggcacg gtggcgctgc cggtcgacaa gcggctgaac cagccggccc cgaagacgca
     4681 gagcgcgctg gccaagtcct tccagaaagc gccgctcgaa acctcggtca ccggttacct
     4741 cgtcaacacc ggctccaagc tggtgctggt ggacaccggc gcggccggcc tgttcggccc
     4801 caccctgggc cggctggcgg ccaacctcaa ggccgcaggc tatcagcccg agcaggtcga
     4861 cgagatctac atcacccaca tgcaccccga ccacgtgggc ggcttgatgg tgggtgagca
     4921 actggcgttc ccgaacgcgg tggtgcgtgc ggaccagaaa gaagccgatt tctggctcag
     4981 ccagaccaac ctcgacaagg ccccggacga cgagagcaaa ggcttcttca aaggcgccat
     5041 ggcctcgctg aacccctatg tgaaggccgg caagttcaag cctttctcgg ggaacaccga
     5101 cctggtgccc ggcatcaaag cgctggccag ccacggccac accccgggcc acaccaccta
     5161 cgtggtcgaa agccaggggc aaaagctcgc cctgctcggc gacctgatac tcgtcgccgc
     5221 ggtgcagttc gacgacccca gcgtcacgaa ccagctcgac atcgacggca agtccgccgc
     5281 ggtggagcgc aagaaggcct tcgcggatgc cgccaagggc ggctacctga tcgcggcgag
     5341 ccacctgccg ttccccggca tcggccacat ccgcgccgaa ggcaagggct accgtttcgt
     5401 gccggtgaac tactcggtcg tcaaccccaa gtgatgatga aagcttggcg taatcatggt
     5461 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5521 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5581 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5641 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5701 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5761 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5821 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5881 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5941 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6001 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6061 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6121 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6181 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6241 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6301 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6361 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6421 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6481 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6541 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6601 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6661 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6721 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6781 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6841 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6901 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6961 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7021 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7081 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7141 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7201 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7261 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7321 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7381 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7441 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7501 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7561 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     7621 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
//
