URL: http://www.addgene.org/24130

Proper Citation: RRID:Addgene_24130

Insert Name: none

Bacterial Resistance: Ampicillin

Defining Citation: PMID:

Vector Backbone Description: Backbone Marker:Didier Trono; Backbone Size:7000; Vector Backbone:pRRL; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin

Comments: pUltra was cloned by Yildirim Dogan (doganym@yahoo.com) This is a 3rd generation Lentiviral vector with an internal Ubiquitinc Promoter. By cloning into the compatible cloning sites* (XbaI and BamHI) downstream of mCherry-P2A, you get a bi-cistronic expression of mCherry and the gene of interest. You can clone a second gene of interest into the NheI/bclI**donwstream of mCherry-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: 3971-4033 and T2A is between bp 4049-4111

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