Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V016968 | p2CT-His-MBP-Lbu_C2c2_R1079A_K1080A | 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:
- p2CT-His-MBP-Lbu_C2c2_R1079A_K1080A
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9434 bp
- Type:
- Gene knockout
- Replication origin:
- ori
- Host:
- E. coli
- Promoter:
- tet
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
p2CT-His-MBP-Lbu_C2c2_R1079A_K1080A vector 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.
p2CT-His-MBP-Lbu_C2c2_R1079A_K1080A vector Sequence
LOCUS V016968 9434 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V016968
VERSION V016968
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 9434)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..9434
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature complement(403..542)
/label="bom"
/note="basis of mobility region from pBR322"
CDS complement(647..835)
/label="rop"
/note="Rop protein, which maintains plasmids at low copy
number"
promoter 2396..2414
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
RBS 2503..2525
/label="RBS"
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 2545..2562
/label="6xHis"
/note="6xHis affinity tag"
CDS 2572..3672
/label="MBP"
/note="maltose binding protein from E. coli"
CDS 3727..3747
/label="TEV site"
/note="tobacco etch virus (TEV) protease recognition and
cleavage site"
CDS 3754..7230
/gene="cas13a"
/label="CRISPR-associated endoribonuclease Cas13a"
/note="CRISPR-associated endoribonuclease Cas13a from
Leptotrichia buccalis (strain ATCC 14201 / DSM 1135 / JCM
12969 / NCTC 10249 / C-1013-b). Accession#: C7NBY4"
terminator 7373..7420
/label="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter complement(7786..7814)
/label="tet promoter"
/note="E. coli promoter for tetracycline efflux protein
gene"
promoter 7927..8031
/label="AmpR promoter"
CDS 8032..8889
/label="AmpR"
/note="beta-lactamase"
rep_origin 9063..9434
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"