Extended Data Fig. 9: The failure of the 0x61 circuit.

The logic diagram is shown with gate colors corresponding to the assigned repressor. The response of the circuit is shown for different combinations of inducer: 100ng/ml aTc, 10mM xylose, and 20mM IPTG. The experimental data are shown as cytometry distributions (black) and blue/red distributions show the ON/OFF output predicted by Cello. The states behave as predicted, except for the -/+/+ state, which should be OFF but is measured as being ON. To determine where this breakage originates the output promoters of intermediate gates are fused to rfp and inserted at the HO locus. The measured responses are then compared to those predicted. The population variability in the response of the CI gate, which causes errors that propagate to the final BM3R1 gate. The DNA sequence of the circuit is provided in Supplementary Table 11, the strains are provided in Supplementary Table 4, and the reporter constructs in Supplementary Table 4. Three experiments were repeated on different days with similar results.