pTet-On vector (V011069)

Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V011069	pTet-On	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

Antibiotic Resistance:: Ampicillin

Length:: 7400 bp

Type:: Mammalian Expression Vectors

Replication origin:: ori

Source/Author:: Gossen M, Freundlieb S, Bender G, Muller G, Hillen W, Bujard H.

Selection Marker:: Neomycin/G418(Geneticin)

Copy Number:: High copy number

Promoter:: SV40

pTet-On vector Vector Map

View Map in NovoBuilder

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 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       pTet-On.        7400 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Vector for expressing the rtTA transactivator in the Tet-On(R) 
            system.
ACCESSION   .
VERSION     .
KEYWORDS    pTet-On
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7400)
  AUTHORS   Gossen M, Freundlieb S, Bender G, Muller G, Hillen W, Bujard H.
  TITLE     Transcriptional activation by tetracyclines in mammalian cells.
  JOURNAL   Science 1995;268:1766-9.
  PUBMED    7792603
REFERENCE   2  (bases 1 to 7400)
  AUTHORS   Clontech
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7400)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science"; 
            date: "1995"; volume: "268"; pages: "1766-9"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     Also known as pUHD17-1neo.
FEATURES             Location/Qualifiers
     source          1..7400
                     /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             774..1778
                     /label=rtTA
                     /note="tetracycline-controlled transactivator, comprising a
                     fusion of the reverse tetracycline repressor rTetR with the
                     C-terminal activation domain of herpes simplex virus VP16"
     polyA_signal    1792..1926
                     /label=SV40 poly(A) signal
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2646..3234)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3408..4265)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4266..4370)
                     /label=AmpR promoter
     polyA_signal    complement(4483..4617)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(5042..5062)
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     intron          complement(5192..5257)
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(5680..6471)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     promoter        complement(6832..7161)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"