LOCUS       Exported                7680 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle expression-secretion vector pP43NMK, complete sequence.
ACCESSION   DQ264732
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7680
                     /organism="Shuttle expression-secretion vector pP43NMK"
                     /lab_host="Bacillus subtilis; Escherichia coli"
                     /mol_type="other DNA"
                     /db_xref="taxon:370428"
     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"
     CDS             1111..1818
                     /codon_start=1
                     /gene="repB"
                     /product="RepB"
                     /label=repB
                     /protein_id="ABD24445.1"
                     /translation="MKHGIQSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSD
                     MAQGFRRMMQYKKINKNLVGFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVN
                     QKQWIQFWKKAMKLDYDPNVKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEK
                     NLKRLSDLEEGLHRKRLISYGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIA
                     MWNWERKNYFIKE"
     gene            1111..1818
                     /gene="repB"
                     /label=repB
     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
                     /regulatory_class="minus_35_signal"
                     /note="-35 and -10 signal of promoter controlled by sigma 
                     B"
     regulatory      4386..4391
                     /regulatory_class="minus_35_signal"
                     /note="-35 and -10 signal of promoter controlled by sigma 
                     A"
     regulatory      4400..4409
                     /regulatory_class="minus_10_signal"
                     /note="-35 and -10 signal of promoter controlled by sigma 
                     B"
     regulatory      4409..4414
                     /regulatory_class="minus_10_signal"
                     /note="-35 and -10 signal of promoter controlled by sigma 
                     A"
     mRNA            4419..5434
                     /gene="mpd"
                     /product="MPH"
                     /label=mpd mRNA
     gene            4419..5434
                     /gene="mpd"
                     /label=mpd
     regulatory      4441..4450
                     /regulatory_class="ribosome_binding_site"
                     /gene="mpd"
     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    5483..5499
                     /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)."
     promoter        complement(5507..5537)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5552..5573
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5678..6469)
                     /direction=LEFT
                     /label=ColE1 ori
                     /note="ColE1 ori"
     rep_origin      complement(5861..6449)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6620..7480)
                     /codon_start=1
                     /gene="bla"
                     /product="Bla"
                     /label=bla
                     /note="beta-galactosidase"
                     /protein_id="ABD24449.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(6620..7480)
                     /gene="bla"
                     /label=bla
     promoter        complement(7481..7585)
                     /gene="bla"
                     /label=AmpR promoter