Fig. 4

Gene copy-number can modulate CAA response profile. a Model simulations predict that gene copy number can skew response profiles. The CAA network topology was stochastically simulated under different circuit copy numbers for A and B. For each copy-number combination, 1000 simulations were performed, and the A and B levels at steady state were compiled to create a pie chart in which green denotes high expression for A and low expression for B, red denotes the reverse, and yellow denotes high expression for both A and B. b Selected strong repression tetO-2x clones were analyzed by flow cytometry to assess circuit expression, and by digital PCR to quantify the number of integrated copies of the two circuit plasmids. The GFP and mCherry genes were used as proxies for their respective plasmids. The native yeast gene ALG9 was used as an endogenous control. Error-bars represent 95% confidence intervals