Basic Vector Information
- Vector Name:
- pVP91A
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6534 bp
- Type:
- Structural Genomics Vectors
- Replication origin:
- ori
- Source/Author:
- Blommel PG, Martin PA, Wrobel RL, Steffen E, Fox BG.
- Copy Number:
- High copy number
- Promoter:
- T5
pVP91A 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 ℃.
pVP91A vector Sequence
LOCUS pVP91A. 6534 bp DNA circular SYN 01-JAN-1980
DEFINITION Bacterial Flexi(R) Vector with an ampicillin resistance marker, for
appending an N-terminal 8xHis tag to an expressed protein.
ACCESSION .
VERSION .
KEYWORDS pVP91A.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6534)
AUTHORS Blommel PG, Martin PA, Wrobel RL, Steffen E, Fox BG.
TITLE High efficiency single step production of expression plasmids from
cDNA clones using the Flexi Vector cloning system.
JOURNAL Protein Expr. Purif. 2006;47:562-70.
PUBMED 16377204
REFERENCE 2 (bases 1 to 6534)
AUTHORS Center for Eukaryotic Structural Genomics
TITLE Direct Submission
REFERENCE 3 (bases 1 to 6534)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Protein
Expr. Purif."; date: "2006"; volume: "47"; pages: "562-70"
COMMENT SGRef: number: 2; type: "Journal Article"
COMMENT To remove the sacB gene, insert an ORF between the SgfI site and the
PmeI site or a downstream site.
FEATURES Location/Qualifiers
source 1..6534
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 126..170
/label=T5 promoter
/note="bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator"
protein_bind 178..194
/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 217..222
/note="ribosome binding site"
CDS 231..233
/codon_start=1
/product="start codon"
/label=start codon
/note="ATG"
/translation="M"
CDS 234..257
/label=8xHis
/note="8xHis affinity tag"
CDS 503..1921
/label=SacB
/note="secreted levansucrase that renders bacterial growth
sensitive to sucrose"
promoter 2010..2040
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 2094..2750
/label=CmR
/note="chloramphenicol acetyltransferase"
misc_feature 2763..2818
/label=MCS
/note="pUC18/19 multiple cloning site"
protein_bind complement(2903..2923)
/label=attB2
/note="core recombination site for the Gateway(R) BP
reaction"
terminator 3010..3096
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
protein_bind complement(3155..3176)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(3192..4271)
/label=lacI
/note="lac repressor"
promoter complement(4272..4349)
/label=lacI promoter
misc_feature 4502..4642
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(4828..5416)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5595..6452)
/label=AmpR
/note="beta-lactamase"
promoter complement(6453..6524)
/label=AmpR promoter
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