Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V006644 | pcDNA6/TR | 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
The pcDNA6⁄TR vector is part of the T-REx System, which yields higher levels of induced expression than any other regulated mammalian expression system. It utilizes the complete CMV promoter and adds control elements from the bacterial tetracycline resistance operon to effectively repress and derepress transcription from one of the strongest mammalian promoter sequences known.The regulatory vector, pcDNA6⁄TR, is provided for high-level expression of the tetracycline repressor (TR) protein. This vector expresses the Blasticidin resistance gene for rapid selection of mammalian cell lines that stably express the TR protein.
- Vector Name:
- pcDNA6/TR
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6662 bp
- Type:
- Tetracycline Regulatory System
- Replication origin:
- ori
- Selection Marker:
- Blasticidin
- Promoter:
- EM7
- Cloning Method:
- Enzyme digestion and ligation
- 5' Primer:
- pCAG-F; Bglob-intron-F
- 3' Primer:
- EBV-rev; BGH-rev
pcDNA6/TR 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.
pcDNA6/TR vector Sequence
LOCUS 40924_10271 6662 bp DNA circular SYN 13-JAN-2022
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6662)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 6662)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6662
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
intron 1028..1600
/label=beta-globin intron
/note="intron from rabbit beta-globin gene"
promoter 1655..1673
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1684..2304
/label=TetR
/note="tetracycline repressor TetR"
polyA_signal 2346..2467
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
CDS 2530..2571
/label=V5 tag
/note="epitope tag from simian virus 5"
CDS 2581..2598
/label=6xHis
/note="6xHis affinity tag"
polyA_signal 2627..2851
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2897..3325
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 3339..3668
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
promoter 3716..3763
/label=EM7 promoter
/note="synthetic bacterial promoter"
CDS 3782..4177
/label=BSD
/note="blasticidin S deaminase"
polyA_signal 4338..4459
/label=SV40 poly(A) signal
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
primer_bind complement(4508..4524)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4532..4548)
/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)."
promoter complement(4556..4586)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4601..4622)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4910..5495)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5669..6526)
/label=AmpR
/note="beta-lactamase"
promoter complement(6527..6631)
/label=AmpR promoter