pTet-IRES-EGFP
(Plasmid #64238)

• Purpose: (Empty Backbone) Lentiviral, doxycycline-inducible system to express transgene of interest and EGFP.
• Depositing Lab: Maria Lung
• Publication: Shuen et al Cancer Gene Ther. 2015 Feb 27. doi: 10.1038/cgt.2015.9.

Backbone

• Vector backbone: pWPI
• Backbone manufacturer: Didier Trono
• Backbone size (bp): 11101
• Modifications to backbone: EF1a promoter from pWPI is removed and replaced with a TetO promoter sequence from pETE-Bsd.
• Vector type: Mammalian Expression, Lentiviral, Cre/Lox
• Promoter: pTet

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Stbl3
• Copy number: High Copy

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: pCEP-F 5’-AGAGCTCGTTTAGTGAACCG-3’
• 3′ sequencing primer: pIRES2-EGFP.P3’ 5’-AACGCACACCGGCCTTATTC-3’

Resource Information

• A portion of this plasmid was derived from a plasmid made by: TetO 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.
• Articles Citing this Plasmid:
  • 5 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

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.

Information for Cloning Grade DNA (Catalog # 64238-DNA.cg) ( Back to top)

Purpose

Cloning grade DNA is suitable for use in PCR, cloning reactions, or transformation into E. coli. The purity and amount is not suitable for direct transfections.

Delivery

• Amount: 2 µg
• Guaranteed Concentration: 100 ng/µl +/- 5 ng/µl
• Pricing: $105 USD
• Storage: DNA can be stored at 4℃ (short term) or -20℃ (long term).

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Quality Control

Addgene has verified this plasmid using Next Generation Sequencing. Results are available here