Basic Vector Information
pnVGW3 vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pnVGW3 vector Sequence
LOCUS 40924_33667 6624 bp DNA circular SYN 18-DEC-2018
DEFINITION Gateway vector pnVGW3 DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6624)
AUTHORS Nishimura K, Ishikawa S, Matsunami E, Yamauchi J, Homma K, Faulkner
C, Oparka K, Jisaka M, Nagaya T, Yokota K, Nakagawa T.
TITLE New Gateway-compatible vectors for a high-throughput protein-protein
interaction analysis by a bimolecular fluorescence complementation
(BiFC) assay in plants and their application to a plant clathrin
structure analysis
JOURNAL Biosci. Biotechnol. Biochem. 79 (12), 1995-2006 (2015)
PUBMED 26193449
REFERENCE 2 (bases 1 to 6624)
AUTHORS Nishimura K, Matsunami E.
TITLE Direct Submission
JOURNAL Submitted (24-FEB-2015) Contact:Kohji Nishimura Shimane University,
Organization of Research; 1060 Nishikawatsu, Matsue, Shimane
690-8504, Japan
REFERENCE 3 (bases 1 to 6624)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6624)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Biosci.
Biotechnol. Biochem."; date: "2015"; volume: "79"; issue: "12";
pages: "1995-2006"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(24-FEB-2015) Contact:Kohji Nishimura Shimane University,
Organization of Research; 1060 Nishikawatsu, Matsue, Shimane
690-8504, Japan"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT constructed using pUC119.
FEATURES Location/Qualifiers
source 1..6624
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 107..128
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 143..173
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 181..197
/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)."
primer_bind 205..221
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 745..1090
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
misc_feature 1111..1740
/note="n-terminal fragment of yellow fluorescent protein
(replacement of gcc with aag at 1729-1731)"
CDS 1111..1629
/codon_start=1
/product="N-terminal fragment of mVenus for use in
bimolecular fluorescence complementation (BiFC) (Kodama and
Hu, 2010)"
/label=VN173
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
ANFKIRHNIE"
protein_bind 1750..1874
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 1911..1941
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 1995..2651
/label=CmR
/note="chloramphenicol acetyltransferase"
CDS 2996..3298
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
protein_bind complement(3342..3466)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
terminator 3492..3744
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(3753..3769)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 3982..4437
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 4719..4823
/label=AmpR promoter
CDS 4824..5681
/label=AmpR
/note="beta-lactamase"
rep_origin 5855..6443
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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