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URL: http://www.addgene.org/217415

Proper Citation: RRID:Addgene_217415

Insert Name: Luciferase, GFP

Organism: Homo sapiens

Bacterial Resistance: Ampicillin

Vector Backbone Description: Backbone Marker:Boris Fehse (Addgene #27358); Vector Backbone:LeGO-iG; Vector Types:Mammalian Expression, Lentiviral, Luciferase; Bacterial Resistance:Ampicillin

Comments: This is an inducible lentiviral luciferase expression vector that has 6 RAS/MAPK-inducible AP-1 sites driving transcription from the minimal CSF2 promoter. LEGO-AP1x6-GM55 was constructed from LEGO-AP1x3-GM55 by duplicating the array of 3 AP-1 sites. LEGO-AP1x3-GM55 was previously constructed from LeGO-iG (Weber et al, 2008) (Addgene #27358) by replacing a NotI/XbaI fragment containing the U6 and SFFV promoters with a fragment from the plasmid pAP1-3 (Addgene #71258) that has 3 copies of the MMP3 gene AP-1 site in front of human CSF2 -55 to +28 minimal promoter driving expression of the Luc+ firefly luciferase gene originating from the plasmid pGL3 (Promega). The luciferase gene is linked to an IRES that in principle supports co-expression of luciferase and EGFP. The insert has 2 copies of the SV40 transcription termination/polyadenylation signal upstream of the multiple cloning site, which blocks background read-through transcription originating in the vector backbone, which itself can be substantial (Bert et al, 2000). The inserted luciferase gene and SV40 polyA site sequences originate from the plasmid pXPG (Bert et al, 2000) (Addgene #71248). References: Bert et al Plasmid. 2000 Sep;44(2):173-82. PMID 10964627. Weber et al Mol Ther. 2008 Apr. 16(4):698-706. PMID 18362927.

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