Fig. 5: Characterization of scaffold for 3-input history-dependent programs.

a Characterization of the final 3-input scaffold and its recombination intermediates DNA states by flow cytometry. We characterized each DNA state by measurement of GFP, RFP, and BFP fluorescence intensities. Each histogram shows fluorescent reporters expressed in the different DNA states, from three experiments with three replicates per experiment. A representative example is depicted here. A detailed design for the final 3-input scaffold and fold change measurements can be found in Fig. S8. b Genetic design of the two plasmids used to implement 3-input programs. The triple controller plasmid regulates the expression of Bxb1, TP901-1, and Int5 integrases. The target plasmid corresponds to the 3-input history-dependent scaffold. c Characterization of a 3-input history-dependent scaffold. We cotransformed the 3-input program with the triple controller plasmid. Bxb1 expression is induced by aTc (input a), Tp901 by arabinose (input b), and Int5 by benzoate (input c). The lineage tree for the program and its corresponding genetic DNA device is represented. For characterizing the system, cells were sequentially induced three times for 16 h each, with different order of occurrences of inputs. Each histogram shows fluorescent reporters expressed in different states. All experiments were performed in triplicate three times on three different days. A representative example is depicted here. Fold change measurements can be found in Fig. S12.