Basic Vector Information
- Vector Name:
- pMCSG18
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6009 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
pMCSG18 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 ℃.
pMCSG18 vector Sequence
LOCUS pMCSG18. 6009 bp DNA circular SYN 01-JAN-1980
DEFINITION Bacterial vector with a 6xHis-TEV leader for high-throughput
purification of C-terminally GFP-tagged proteins.
ACCESSION .
VERSION .
KEYWORDS pMCSG18.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6009)
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 6009)
AUTHORS Midwest Center for Structural Genomics
TITLE Direct Submission
REFERENCE 3 (bases 1 to 6009)
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..6009
/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(214..921)
/label=GFP
/note="Aequorea victoria green fluorescent protein"
CDS complement(946..966)
/label=TEV site
/note="tobacco etch virus (TEV) protease recognition and
cleavage site"
CDS complement(991..1008)
/label=6xHis
/note="6xHis affinity tag"
CDS complement(1009..1011)
/codon_start=1
/product="start codon"
/label=start codon
/note="ATG"
/translation="M"
RBS complement(1019..1041)
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind complement(1056..1080)
/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(1081..1099)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 1412..1489
/label=lacI promoter
CDS 1490..2569
/label=lacI
/note="lac repressor"
protein_bind 2585..2606
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS 3381..3569
/label=rop
/note="Rop protein, which maintains plasmids at low copy
number"
misc_feature 3674..3816
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(4002..4590)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4764..5621)
/label=AmpR
/note="beta-lactamase"
promoter complement(5622..5725)
/label=AmpR promoter
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