Replication-deficient Ad5 with SV40-Renilla luciferase expression cassette in place of the E1 region

Ad291-RL is an Ad5-based adenovirus containing a 1.8 kb-long Renilla luciferase expression cassette in place of the E1 region, oriented from left to right relative to the adenovirus genome.  The Renilla luciferase coding sequence is under the control of a SV40 enhancer/early promoter and a SV40 late polyadenylation signal.  Both E1 (3.1 kb) and E3 (2.8 kb) regions are deleted.  The two SfiI sites naturally present in WT Ad5 DNA were mutated by substituting A for G and C at positions 16291 and 16294 in the Ad5 genome, and C and G for respectively G and C at positions 23001 and 23004 in the Ad5 genome, introducing silent mutations in the adenovirus pVII and DNA-binding protein coding sequences.