Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012519 | pEX-N-His-GST | In stock, 1 week for quality controls |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pEX-N-His-GST
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5227 bp
- Type:
- Cloning Vectors
- Replication origin:
- ori
- Source/Author:
- OriGene
- Copy Number:
- High copy number
pEX-N-His-GST vector Map
Plasmid 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 ℃.
6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it
General Plasmid Transform Protocol
1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.
pEX-N-His-GST vector Sequence
LOCUS pEX-N-His-GST. 5227 bp DNA circular SYN 01-JAN-1980
DEFINITION PrecisionShuttle(TM) bacterial vector for inducible expression of a
protein with a cleavable N-terminal 6xHis-GST tag.
ACCESSION .
VERSION .
KEYWORDS pEX-N-His-GST
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5227)
AUTHORS OriGene
TITLE Direct Submission
REFERENCE 2 (bases 1 to 5227)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT Inserts are typically cloned between the SgfI (AsiSI) and MluI
sites.
FEATURES Location/Qualifiers
source 1..5227
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 155..171
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 207..225
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 226..250
/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)."
RBS 254..276
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 284..286
/codon_start=1
/product="start codon"
/label=start codon
/note="ATG"
/translation="M"
CDS 287..304
/label=6xHis
/note="6xHis affinity tag"
CDS 311..964
/label=GST
/note="glutathione S-transferase from Schistosoma
japonicum"
CDS 986..1006
/label=TEV site
/note="tobacco etch virus (TEV) protease recognition and
cleavage site"
misc_feature 1007..1093
/label=MCS
/note="MCS"
/note="multiple cloning site"
misc_feature 1060..1062
/label=stop codon
/note="stop codon"
terminator 1132..1178
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter 1424..1501
/label=lacI promoter
CDS 1502..2581
/label=lacI
/note="lac repressor"
protein_bind 2597..2618
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2633..2663
/label=lac promoter
/note="promoter for the E. coli lac operon"
primer_bind 2695..2711
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2727..2743)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin complement(3035..3623)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3797..4654)
/label=AmpR
/note="beta-lactamase"
promoter complement(4655..4759)
/label=AmpR promoter
rep_origin 4786..5227
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"