Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012830 | pTRE2hyg-Stuffer | In stock, instant shipping |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pTRE2hyg-Stuffer
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6966 bp
- Type:
- Mammalian Expression Vectors
- Replication origin:
- ori
- Selection Marker:
- Hygromycin
- Copy Number:
- High copy number
- Promoter:
- SV40
- Cloning Method:
- Enzyme Cut
- Fusion Tag:
- Luc
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
pTRE2hyg-Stuffer 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.
pTRE2hyg-Stuffer vector Sequence
LOCUS V012830 6966 bp DNA circular SYN 13-DEC-2023
DEFINITION Exported.
ACCESSION V012830
VERSION V012830
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 6966)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..6966
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 15..285
/label="tetracycline response element"
/note="contains seven copies of the tetracycline operator
tetO"
promoter 318..356
/label="minimal CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 480..2129
/label="luciferase"
/note="firefly luciferase"
intron 2195..2767
/label="beta-globin intron"
/note="intron from rabbit beta-globin gene"
CDS 2771..2893
/gene="HBB"
/label="Hemoglobin subunit beta"
/note="Hemoglobin subunit beta from Colobus guereza.
Accession#: Q9TT33"
polyA_signal 2964..3019
/label="beta-globin poly(A) signal"
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
rep_origin complement(3581..4169)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4343..5200)
/label="AmpR"
/note="beta-lactamase"
promoter complement(5201..5305)
/label="AmpR promoter"
polyA_signal complement(5406..5487)
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
CDS complement(5607..6629)
/label="HygR"
/note="aminoglycoside phosphotransferase from E. coli"
promoter complement(join(6638..6966,1))
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"