Loss-of-function mutations and evolutionary stability dynamics in re-engineered I7101 circuits (A) I7101 (R0011 + E0240) re-engineering involves swapping out the R0011 promoter. (B) Evolutionary stability dynamics of R0011 + E0240 (open black circles) and re-engineered circuits (various shapes and colors) under constitutive expression. Error bars represent one standard deviation from the mean of nine independently evolved populations. (C) Types of mutations in nine independently evolved populations. For nine independently evolved populations, colored boxes correspond to the mutation legend below the table. The most common mutation for a particular type of mutation is labeled with "1" in the boxes above and less common mutations are labeled with increasing numbers. (D) Most common loss-of-function mutations that inactivated the re-engineered I7101 circuits. See Additional File 1, Supplementary Table S1 for mutation details.