Mutational trajectories and fitness parameters during adaptive evolution. Adaptive trajectory of strain BW25113(Δpts, Δglk) as measured by reconstitution of the (A) glucokinase phenotype with associated mutations and (B) fitness measured as growth rates. (C, D) Strains BW25113(Δpts, Δglk, Δmak) and (E, F) BW25113(Δpts::kanR, Δglk) display a similar adaptive trajectory. Mutations observed from whole-genome sequencing are annotated at the respective generation when they appear at 20% or greater frequency in the population. Points represent averages (n = 3) ± SD. Dashed lines indicate corresponding parameters for strain M9.2.50.