Basic Vector Information
- Vector Name:
- pMCSG38C
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6313 bp
- Type:
- Structural Genomics Vectors
- Replication origin:
- ori
- Source/Author:
- Stols L, Gu M, Dieckman L, Raffen R, Collart FR, Donnelly MI.
- Copy Number:
- High copy number
pMCSG38C 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 ℃.
pMCSG38C vector Sequence
LOCUS pMCSG38C. 6313 bp DNA circular SYN 01-JAN-1980
DEFINITION Bacterial expression vector with a 6xHis-TEV leader and a
Tet-inducible TVMV protease cassette. See also pMCSG38.
ACCESSION .
VERSION .
KEYWORDS pMCSG38C.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6313)
AUTHORS Stols L, Gu M, Dieckman L, Raffen R, Collart FR, Donnelly MI.
TITLE A new vector for high-throughput, ligation-independent cloning
encoding a tobacco etch virus protease cleavage site.
JOURNAL Protein Expr. Purif. 2002;25:8-15.
PUBMED 12071693
REFERENCE 2 (bases 1 to 6313)
AUTHORS Midwest Center for Structural Genomics
TITLE Direct Submission
REFERENCE 3 (bases 1 to 6313)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Protein
Expr. Purif."; date: "2002"; volume: "25"; pages: "8-15"
COMMENT SGRef: number: 2; type: "Journal Article"
COMMENT For ligation-independent cloning (LIC), linearize with SspI and
treat with T4 DNA polymerase plus dGTP.
FEATURES Location/Qualifiers
source 1..6313
/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(223..243)
/label=TEV site
/note="tobacco etch virus (TEV) protease recognition and
cleavage site"
CDS complement(268..285)
/label=6xHis
/note="6xHis affinity tag"
CDS complement(286..288)
/codon_start=1
/product="start codon"
/label=start codon
/note="ATG"
/translation="M"
RBS complement(296..318)
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind complement(333..357)
/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(358..376)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 675..748
/label=PLtetO-1 promoter
/note="modified phage lambda PL promoter with tet operator
sites (Lutz and Bujard, 1997)"
CDS 772..1479
/label=TVMV protease
/note="tobacco vein mottling virus NIa protease
(Nallamsetty et al., 2004)"
terminator 1515..1601
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
promoter 1716..1793
/label=lacI promoter
CDS 1794..2873
/label=lacI
/note="lac repressor"
protein_bind 2889..2910
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS 3685..3873
/label=rop
/note="Rop protein, which maintains plasmids at low copy
number"
misc_feature 3978..4120
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(4306..4894)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5068..5925)
/label=AmpR
/note="beta-lactamase"
promoter complement(5926..6029)
/label=AmpR promoter
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