Figure 1

The poxvirus ablation /insertion vector p2KO. Separate but identical poxvirus early/late synthetic promoters allowed for simple and targeted ablation of selected gene(s) with the simultaneous expression of the fluorescent reporter mCherry with or without the expression of an additional expressed transgene. The left and right flanks are bounded by pairs of unique restriction sites (Sac I and Not I for the left flank, EcoR I and Hind III for the right flank). The expressed transgene ORF is bounded by unique a 5′- BamH I and a 3′- Xma I restriction site. These allow for the simple and directional ligation of PCR amplicons bounded by the appropriate restriction sites. Numbers in small type refer to nucleotide positions within the p2KO vector.