I find it interesting that before the removal of the circuit, the RQ70 strain was slightly slower than the MG70 control. Do you think that's from the burden of the circuit or the format of the culture that you see making a large difference later in the results, or a mixture of the two? Since the culture format changed the rankings between the strains so much, I wonder how much the format of the culture during the selection would change the results. Excited to see if you look into the specific genetic changes you saw in RQ70 in the future!

Fascinating study! Have you looked at whether the position of the deoxycytidine substitutions matters? For example, do substitutions near the 5′ end have a different effect than those near the 3′ end, or is it mainly the total amount of dCTP incorporation that drives the phenotype? The differences between the luciferase and eGFP reporters in Figure 2 also made me wonder whether the effect depends on the transcript's underlying sequence composition.