Purpose(Empty Backbone) Lentiviral, doxycycline-inducible system to express transgene of interest and EGFP.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||64238||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
Backbone manufacturerDidier Trono
- Backbone size (bp) 11101
Modifications to backboneEF1a promoter from pWPI is removed and replaced with a TetO promoter sequence from pETE-Bsd.
Vector typeMammalian Expression, Lentiviral, Cre/Lox
- Promoter pTet
Growth in Bacteria
Copy numberHigh Copy
- Cloning method Restriction Enzyme
- 5′ sequencing primer pCEP-F 5’-AGAGCTCGTTTAGTGAACCG-3’
- 3′ sequencing primer pIRES2-EGFP.P3’ 5’-AACGCACACCGGCCTTATTC-3’ (Common Sequencing Primers)
A portion of this plasmid was derived from a plasmid made byTetO promoter is from pETE-Bsd vector and the vector backbone is from pWPI vector. The pETE-BSD was obtained from Prof Eugene Zabarovsky around 1996. The pWPI vector was obtained from Prof Didier Trono in 2009.
Terms and Licenses
The transgene of interest can be inserted into BamHI, PmeI, and PstI sites. However, only the PmeI and PstI sites are recommended to be used for different restriction end ligation.
This plasmid must be used together with an independent tTA-expressing plasmid for Tet-Off system or rtTA-expressing plasmid for Tet-On system.
The TetO promoter sequence was excised from pETE-Bsd plasmid vector, which was originated from pUHD-10-3. The pETE-Bsd and pUHD-10-13 were engineered by Protopopov et al. and Damke et al, respectively.
Protopopov AI, Li J, Winberg G, Gizatullin RZ, Kashuba VI, Klein G, et al. Human cell lines engineered for tetracycline-regulated expression of tumor suppressor candidate genes from a frequently affected chromosomal region, 3p21. J Gene Med. 2002/07/19 ed. 2002;4(4):397–406.
Damke, H., Gossen, M., Freundlieb, S., Bujard, H., and Schmid, S.L. (1995). Tightly regulated and inducible expression of dominant interfering dynamin mutant in stably transformed HeLa cells. Methods Enzymol. 257, 209–220.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:pTet-IRES-EGFP was a gift from Maria Lung (Addgene plasmid # 64238 ; http://n2t.net/addgene:64238 ; RRID:Addgene_64238)
For your References section:Novel lentiviral-inducible transgene expression systems and versatile single-plasmid reporters for in vitro and in vivo cancer biology studies. Shuen WH, Kan R, Yu Z, Lung HL, Lung ML. Cancer Gene Ther. 2015 Feb 27. doi: 10.1038/cgt.2015.9. 10.1038/cgt.2015.9 PubMed 25721206