Cheater Frequency Test

Single colonies of the Δung::FRT ΔpheA::FRT Δflu::AtPAT strain harboring pHyo182-AtADT2 and pHyo094 (T7RNAP-CD fusion or T7RNAP alone) were inoculated into 3 mL of LB medium (Carb and Cmp) and incubated overnight. Cultures were diluted 100-fold into LB (Carb, Cmp, 0.1 mM IPTG, 0.2% arabinose), grown for 4 h, and subcultured twice more. Cells were pelleted, washed, resuspended, inoculated into MOPS, and grown as above. Cultures were then diluted to OD₆₀₀ 1, and 100 μL aliquots were plated on MOPS medium (Carb, Cmp, 1 mM IPTG, 0.002% arabinose) containing 5 mM 4FPA; colonies were counted after 4 days. Aliquots were also diluted 10⁵-fold and plated on LB (Carb and Cmp) for total viable cell counts.