pWM91 vector (V002128)

Basic Vector Information

Vector Name:
pWM91
Antibiotic Resistance:
Ampicillin
Length:
8261 bp
Type:
Cloning vector
Replication origin:
R6K γ ori
Source/Author:
Okamoto S, Niki H.
Promoter:
sacB

pWM91 vector Map

pWM918261 bp400800120016002000240028003200360040004400480052005600600064006800720076008000R6K gamma oriM13 fwdT7 promoterMCST3 promoterM13 revlac operatorlac promoterCAP binding siteoriTbacterial terminatorincP origin of transferhis operon terminatororisacB promoterSacBAmpRAmpR promoter

pWM91 vector Sequence

LOCUS       40924_46593        8261 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pWM91 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8261)
  AUTHORS   Okamoto S, Niki H.
  TITLE     cloning vector pWM91 sequence
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8261)
  AUTHORS   Okamoto S, Niki H.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-APR-2016) Contact:Sho Okamoto National Institute of 
            Genetics, Microbial Genetics Laboratory, Genetics Strains Research 
            Center; 1111 Yata, Mishima, Shizuoka 411-8540, Japan URL 
            :https://www.nig.ac.jp/labs/MicroGen/
REFERENCE   3  (bases 1 to 8261)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8261)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (01-APR-2016) Contact:Sho Okamoto National Institute of Genetics, 
            Microbial Genetics Laboratory, Genetics Strains Research Center; 
            1111 Yata, Mishima, Shizuoka 411-8540, Japan URL 
            :https://www.nig.ac.jp/labs/MicroGen/"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8261
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(59..415)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     primer_bind     746..762
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        772..790
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(799..906)
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(919..937)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(958..974)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(982..998)
                     /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(1006..1036)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1051..1072)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     oriT            1778..1887
                     /label=oriT
                     /note="incP origin of transfer"
     terminator      complement(2113..2156)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     oriT            2208..2317
                     /label=incP origin of transfer
                     /note="incP origin of transfer"
     terminator      2565..2624
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E.
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     rep_origin      complement(2819..3407)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        4444..4889
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             4890..6308
                     /codon_start=1
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVSEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     CDS             complement(7199..8056)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8057..8161)
                     /label=AmpR promoter

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