Basic Vector Information
pVP13 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 ℃.
pVP13 vector Sequence
LOCUS pVP13. 7675 bp DNA circular SYN 01-JAN-1980
DEFINITION Bacterial Gateway(R) destination vector for appending an N-terminal
S-Tag-6xHis-MBP cassette to an expressed protein. Also known as
pVP13-GW.
ACCESSION .
VERSION .
KEYWORDS pVP13.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7675)
AUTHORS Thao S, Zhao Q, Kimball T, Steffen E, Blommel PG, Riters M, Newman
CS, Fox BG, Wrobel RL.
TITLE Results from high-throughput DNA cloning of Arabidopsis thaliana
target genes using site-specific recombination.
JOURNAL J. Struct. Funct. Genomics 2004;5:267-76.
PUBMED 15750721
REFERENCE 2 (bases 1 to 7675)
AUTHORS Center for Eukaryotic Structural Genomics
TITLE Direct Submission
REFERENCE 3 (bases 1 to 7675)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J. Struct.
Funct. Genomics"; date: "2004"; volume: "5"; pages: "267-76"
COMMENT SGRef: number: 2; type: "Journal Article"
COMMENT A TEV cleavage site can be encoded during PCR amplification.
FEATURES Location/Qualifiers
source 1..7675
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 11..55
/label=T5 promoter
/note="bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator"
protein_bind 63..79
/label=lac repressor encoded by lacI binding site
/bound_moiety="lac repressor encoded by lacI"
/note="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)."
RBS 102..107
/note="ribosome binding site"
CDS 116..118
/codon_start=1
/product="start codon"
/label=start codon
/note="ATG"
/translation="M"
CDS 122..166
/label=S-Tag
/note="affinity and epitope tag derived from pancreatic
ribonuclease A"
CDS 173..190
/label=6xHis
/note="6xHis affinity tag"
CDS 194..1291
/label=MBP
/note="maltose binding protein from E. coli"
protein_bind 1361..1485
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 1510..1540
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 1594..2250
/label=CmR
/note="chloramphenicol acetyltransferase"
CDS 2595..2897
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
protein_bind complement(2941..3065)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
terminator 3133..3227
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS 3271..3927
/label=CmR
/note="chloramphenicol acetyltransferase"
terminator 3995..4081
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
protein_bind complement(4140..4161)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(4177..5256)
/label=lacI
/note="lac repressor"
promoter complement(5257..5334)
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
misc_feature 5526..5666
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(5852..6440)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6614..7471)
/label=AmpR
/note="beta-lactamase"
promoter complement(7472..7576)
/label=AmpR promoter
This page is informational only.