|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||14901||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
Backbone manufacturerVerma Lab
- Backbone size w/o insert (bp) 8600
Vector typeMammalian Expression, Lentiviral
Growth in Bacteria
Copy numberLow Copy
Alt nameCMV GFP
Insert Size (bp)3000
- Cloning method Restriction Enzyme
- 5′ cloning site NheI (destroyed during cloning)
- 3′ cloning site Eco47 (destroyed during cloning)
- 5′ sequencing primer n/a (Common Sequencing Primers)
Terms and Licenses
This tet-inducible vector was constructed by cloning an XbaI/HpaI fragment containing the tTA and the
inducible promoter from pBStind into NheI/Eco47(AfeI) sites of pCL-CG. The inducible cassette contains the tet dependent
transactivator (tTA) at its 5' end and an
inducible promoter at its 3' end. Expression
of the GFP gene is controlled by the inducible promoter,
which contains a minimal promoter (mp) and seven
copies of Tet operator (tetO). The tTA gene is constitutively
expressed under the control of the CMV promoter.
Following transduction, the constitutively
expressed tTA (tetRVP16) binds to tet(O)- mp and
induces the transcription of a high level of GFP mRNA.
In contrast, binding of doxycycline to the tTA induces
conformational changes that prevent it from binding to
the tet(O)- minimal promoter and consequently leads to
little or no GFP expression.
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:pCL-CTIG was a gift from Inder Verma (Addgene plasmid # 14901 ; http://n2t.net/addgene:14901 ; RRID:Addgene_14901)
For your References section:Lentiviral vectors: regulated gene expression. Kafri T, van Praag H, Gage FH, Verma IM. Mol Ther. 2000 Jun . 1(6):516-21. 10.1006/mthe.2000.0083 PubMed 10933976