Purpose(Empty Backbone) 3rd generation Lentiviral vector for bi-cistronic expression of EGFP and the gene of interest
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||24129||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) 7000
Vector typeMammalian Expression, Lentiviral
Growth in Bacteria
Growth instructionsStbl3 or SC110 (See below**)
/ Fusion Protein
- EGFP (N terminal on backbone)
- Cloning method Restriction Enzyme
- 5′ cloning site XbaI/NheI (not destroyed)
- 3′ cloning site BamHI/BcII (not destroyed)
- 5′ sequencing primer hUBCpro-F (Common Sequencing Primers)
Terms and Licenses
Articles Citing this Plasmid
pUltra was cloned by Yildirim Dogan ([email protected]). It is described in the associated article (PMID 22427958) and also in PMID 24630793.
This is a 3rd generation Lentiviral vector with an internal Ubiquitinc Promoter.
By cloning into the compatible cloning sites* (XbaI and BamHI) downstream of EGFP-P2A, you get a bi-cistronic expression of EGFP and the gene of interest. You can clone a second gene of interest into the NheI/bclI**donwstream of EGFP-P2A-gene1-T2A and get a multi-cistronic expression of all three genes.
* compatible cloning site (ccs): you can pcr amplifiy your gene of interest with a forward primer including one of these cutting sites: SpeI or NheI or XbaI and a reverse primer with one these cutting sites: BglII or BamHI or bclI. The PCR product is now compatible with the first site as well the 2nd site. "You can mix and match".
** for bclI the vector has to be amplified by dam-methylation defective E coli strains (e.g. SC110 Stratagene).
Further advantage of the vector is, that you can simultaneously do RNAi by cloning H1-shRNA cassettes into the unique SnaBI site in the 3´-LTR. Plus while integration the RNAi cassette gets doubled, since the 5´LTR is dublicated while reverse transcripton from the 3´-LTR
***P2A: 3984-4046 bp and T2A: 4062-4124 bp
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:pUltra was a gift from Malcolm Moore (Addgene plasmid # 24129 ; http://n2t.net/addgene:24129 ; RRID:Addgene_24129)
For your References section:Tunneling nanotubes provide a unique conduit for intercellular transfer of cellular contents in human malignant pleural mesothelioma. Lou E, Fujisawa S, Morozov A, Barlas A, Romin Y, Dogan Y, Gholami S, Moreira AL, Manova-Todorova K, Moore MA. PLoS One. 2012;7(3):e33093. doi: 10.1371/journal.pone.0033093. Epub 2012 Mar 9. 10.1371/journal.pone.0033093 PubMed 22427958