Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V011070 | pTet-On-Advanced | 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:
- pTet-On-Advanced
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7139 bp
- Type:
- Mammalian Expression Vectors
- Replication origin:
- ori
- Source/Author:
- Clontech
- Selection Marker:
- Neomycin/G418(Geneticin)
- Copy Number:
- High copy number
- Promoter:
- SV40
pTet-On-Advanced 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.
pTet-On-Advanced vector Sequence
LOCUS pTet-On-Advanced 7139 bp DNA circular SYN 01-JAN-1980
DEFINITION Vector for expressing the rtTA-Advanced transactivator in the
Tet-On(R) Advanced system.
ACCESSION .
VERSION .
KEYWORDS pTet-On-Advanced
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7139)
AUTHORS Clontech
TITLE Direct Submission
REFERENCE 2 (bases 1 to 7139)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7139
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 89..468
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 469..672
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 775..1518
/label=rtTA-Advanced
/note="improved tetracycline-controlled transactivator"
polyA_signal 1531..1665
/label=SV40 poly(A) signal
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin complement(2385..2973)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3147..4004)
/label=AmpR
/note="beta-lactamase"
promoter complement(4005..4109)
/label=AmpR promoter
polyA_signal complement(4222..4356)
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
CDS complement(4781..4801)
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
intron complement(4931..4996)
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS complement(5419..6210)
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
promoter complement(6571..6900)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"