Price Information
Cat No. | Plasmid Name | Availability | Add to cart |
---|---|---|---|
V011045 | pTRE3G | 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:
- pTRE3G
- Antibiotic Resistance:
- Ampicillin
- Length:
- 3431 bp
- Type:
- Mammalian Expression Vectors
- Replication origin:
- ori
- Source/Author:
- Clontech
- Copy Number:
- High copy number
- Promoter:
- TRE3G
pTRE3G 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.
pTRE3G vector Sequence
LOCUS pTRE3G. 3431 bp DNA circular SYN 01-JAN-1980 DEFINITION Vector for doxycycline-inducible expression of a gene in cells containing the Tet-On(R) 3G transactivator protein. ACCESSION . VERSION . KEYWORDS pTRE3G SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3431) AUTHORS Clontech TITLE Direct Submission REFERENCE 2 (bases 1 to 3431) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..3431 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 4..382 /label=TRE3G promoter /note="3rd-generation Tet-responsive promoter that can be activated by binding of Tet-On(R) 3G" misc_feature 383..452 /label=MCS /note="MCS" /note="multiple cloning site" intron 583..648 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS 778..798 /label=SV40 NLS /note="nuclear localization signal of SV40 (simian virus 40) large T antigen" polyA_signal 1223..1344 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin complement(1601..2189) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(2363..3220) /label=AmpR /note="beta-lactamase" promoter complement(3221..3325) /label=AmpR promoter