URL: http://www.addgene.org/14884
Proper Citation: RRID:Addgene_14884
Insert Name: flap-Ub promoter-GFP-WRE
Bacterial Resistance: Ampicillin
Defining Citation: PMID:12518064
Vector Backbone Description: Backbone Marker:I. Verma, Salk; Backbone Size:0; Vector Backbone:HR'CS-G; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Comments: FG12 was derived from FUGW. The extra nucleotides from the HindIII site downstream of the Ubiquitin-C promoter (UbiC) promoter to the NcoI site in front of the initiation codon of EGFP were deleted by a HindIII-NcoI adapter ligation. Further, XbaI, EcoRI, and XhoI sites at the 3' end of EGFP and WRE were eliminated, followed by a polylinker oligonucleotide ligation at the PacI site between the Flap element and the UbiC promoter, to generate a set of new restriction sites, XbaI-HpaI-XhoI-BstXI-PacI, that is optimal for accommodating the siRNA expression cassette. To construct the siRNA-expressing lentiviral vectors, the siRNA expression cassette can be subcloned into FG12 between the XbaI and XhoI sites. Lentiviruses can be produced with third generation packaging system plasmids pMDLg/pRRE, pRSV-Rev, and pMD2.G.
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Source: Addgene