Replication-deficient Ad5 with SV40-Renilla luciferase expression cassette in place of the E1 region and SV40-Firefly luciferase in place of the E3 region

Ad292-RL-FL is an Ad5-based adenovirus that contains a 1.8 kb-long Renilla luciferase expression cassette in place of the E1 region, and a 2.2 kb-long firefly luciferase expression cassette in place of the E3 region. The luciferase coding sequences are both under the control of a SV40 enhancer/early promoter and a SV40 late polyadenylation signal.  They are oriented from left to right relative to the adenovirus genome.  The sizes of the E1 and E3 deletions are 3.1 kb and 2.8 kb, respectively.  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.