(A) Bxz2 Int-mediated recombination in vitro between linear attB (pSS1 linearized with BamH1) and 60 bp attP DNA substrates; Int concentrations are: 0, 16, 33, 66, 125, 250, 500, 1000 nM. (B) Peaches-Int-mediated recombination in vitro. Peaches-Int concentrations: 0, 125, 250, 500, 600, 700, 1000, 2000 nM. (C) In vitro integration varying temperature (°C); Peaches-Int concentration, 700 nM. (D, E) Sizes of synthetic Bxz2 attP and M. smegmatis attB substrates used to determine site requirements, and (F, G) determination of their recombination activities with Bxz2 Int. The sizes of the attP (F) and attB (G) substrates are shown above each lane. Positions of attL and attR products are indicated.