|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||12246||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 12290
Vector typeMammalian Expression, Lentiviral, Cre/Lox
Growth in Bacteria
Growth instructionsUse Stbl3 or HB101 to reduce chance of recombination. Grow at 37C
Copy numberHigh Copy
Gene/Insert nameSV40 Large T, HSV1-TK
Alt nameSV40 Large T
Alt nameHSV-1 thymidine kinase
MutationLoxP in 3' LTR.
Entrez GeneSV40gp6 (a.k.a. SV40gp6)
- Cloning method Restriction Enzyme
- 5′ sequencing primer CMV Fwd for LargeT
- 3′ sequencing primer SP6 primer rev for LoxP (Common Sequencing Primers)
CMV promoter is upstream of the bicistronic coding cassette with SV40-Large T and HSV1-TK. During reverse transcription U3 region of the 3'LTR, with LoxP, is duplicated; LoxP sites end up flanking the genome of the integrated provirus. Upon expression of Cre, provirus is excised.
Please note that the full sequence for this plasmid is approximated and not fully verified--there will be discrepancies between Addgene's quality control sequence and the theoretical sequence from the depositor. Please visit the Trono lab http://tronolab.epfl.ch for cloning strategies, protocols, publications, and more. See LentiWeb http://www.lentiweb.com for discussion on cloning strategies and protocols.
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:pLOX-Ttag-iresTK was a gift from Didier Trono (Addgene plasmid # 12246 ; http://n2t.net/addgene:12246 ; RRID:Addgene_12246)
For your References section:Reversible immortalization of human primary cells by lentivector-mediated transfer of specific genes. Salmon P, Oberholzer J, Occhiodoro T, Morel P, Lou J, Trono D. Mol Ther. 2000 Oct . 2(4):404-14. 10.1006/mthe.2000.0141 PubMed 11020357