LOCUS       Exported                6841 bp DNA     circular SYN 28-OCT-2024
DEFINITION  Shuttle expression-secretion vector pP43NMK, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6841)
  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 6841)
  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 6841)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6841)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 6841)
  TITLE     Direct Submission
REFERENCE   6  (bases 1 to 6841)
  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..6841
                     /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..3369
                     /codon_start=1
                     /gene="ble"
                     /product="Ble"
                     /label=ble
                     /protein_id="ABD24447.1"
                     /translation="MRMLQSIPALPVGDIKKSIGFYCDKLGFTLVHHEDGFAVLMCNEV
                     RIHLWEASDEGWRSRSNDSPVCTGAESFIAGTASCRIEVEGIDELYQHIKPLGILHPNT
                     SLKDQWWDERDFAVIDPDNNLISFFQQIKS"
     gene            2965..3217
                     /gene="ble"
                     /label=ble
     promoter        3589..3808
                     /label=Hpa II promoter
     RBS             3859..3867
     CDS             3876..4589
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(4620..4636)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4644..4660)
                     /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(4668..4698)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4713..4734)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5022..5610)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5784..6641)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6642..6746)
                     /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 taaaggagaa gaacttttca
     3901 ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg
     3961 tcagtggaga gggtgaaggt gatgcaacat acggaaaact tacccttaaa tttatttgca
     4021 ctactggaaa actacctgtt ccatggccaa cacttgtcac tactttcggt tatggtgttc
     4081 aatgctttgc gagataccca gatcatatga aacagcatga ctttttcaag agtgccatgc
     4141 ccgaaggtta tgtacaggaa agaactatat ttttcaaaga tgacgggaac tacaagacac
     4201 gtgctgaagt caagtttgaa ggtgataccc ttgttaatag aatcgagtta aaaggtattg
     4261 attttaaaga agatggaaac attcttggac acaaattgga atacaactat aactcacaca
     4321 atgtatacat catggcagac aaacaaaaga atggaatcaa agttaacttc aaaattagac
     4381 acaacattga agatggaagc gttcaactag cagaccatta tcaacaaaat actccaattg
     4441 gcgatggccc tgtcctttta ccagacaacc attacctgtc cacacaatct gccctttcga
     4501 aagatcccaa cgaaaagaga gaccacatgg tccttcttga gtttgtaaca gctgctggga
     4561 ttacacatgg catggatgaa ctatacaaat aaatgctgca gaagcttggc gtaatcatgg
     4621 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4681 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     4741 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4801 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4861 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4921 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4981 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5041 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5101 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5161 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5221 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5281 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5341 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5401 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5461 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5521 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5581 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5641 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5701 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5761 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5821 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5881 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5941 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6001 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6061 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6121 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6181 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6241 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6301 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6361 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6421 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6481 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6541 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6601 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6661 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6721 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     6781 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     6841 c
//
