Basic Vector Information
- Vector Name:
- pMCSG43
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6773 bp
- Type:
- Structural Genomics Vectors
- Replication origin:
- ori
- Source/Author:
- Midwest Center for Structural Genomics
- Copy Number:
- High copy number
- Promoter:
- dsbA
pMCSG43 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 ℃.
pMCSG43 vector Sequence
LOCUS V010655 6773 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V010655
VERSION V010655
KEYWORDS pMCSG43.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 6773)
AUTHORS Midwest Center for Structural Genomics
TITLE Direct Submission
REFERENCE 2 (bases 1 to 6773)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
For ligation-independent cloning (LIC), linearize with SmaI and
treat with T4 DNA polymerase plus dATP.
FEATURES Location/Qualifiers
source 1..6773
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator complement(26..73)
/label="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(140..157)
/label="6xHis"
/note="6xHis affinity tag"
CDS complement(208..225)
/label="6xHis"
/note="6xHis affinity tag"
CDS complement(232..252)
/label="TEV site"
/note="tobacco etch virus (TEV) protease recognition and
cleavage site"
CDS complement(263..283)
/label="TVMV site"
/note="tobacco vein mottling virus (TVMV) NIa protease
recognition and cleavage site"
CDS complement(305..907)
/gene="osmY"
/label="Osmotically-inducible protein Y"
/note="Osmotically-inducible protein Y from Escherichia
coli (strain K12). Accession#: P0AFH8"
RBS complement(915..937)
/label="RBS"
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind complement(952..976)
/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(977..995)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 1213..1444
/label="dsbA promoter"
/note="promoter for the E. coli dsbA gene (Belin and
Boquet, 1994)"
CDS 1445..2068
/label="DsbA"
/note="bacterial periplasmic oxidoreductase"
promoter 2176..2253
/label="lacI promoter"
CDS 2254..3333
/label="lacI"
/note="lac repressor"
protein_bind 3349..3370
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
CDS 4145..4333
/label="rop"
/note="Rop protein, which maintains plasmids at low copy
number"
misc_feature 4438..4580
/label="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(4766..5354)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5528..6385)
/label="AmpR"
/note="beta-lactamase"
promoter complement(6386..6489)
/label="AmpR promoter"
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