Figure 2.

Construction of a complex logic gate and oscillator. (A) Using ApE, one can take a sequence of parts from a design, assign to one or more vectors, and edit any parts or junctions at the DNA sequence level. (B) Raven is used to determine a hierarchical assembly plan for each construct using a specific assembly method. Assembling an AND gate with Gibson requires two stages, four steps, four synthesis steps, and six polymerase chain reaction (PCR) steps, whereas assembling the oscillator constructs with GoldenGate requires two stages, four steps, and 16 PCR steps. Raven generates oligonucleotides to perform these assemblies, but j5 can be used for more refined primer designs for specific cloning steps. The common sequences are highlighted in red. (C) Once a complete assembly plan is determined, PR-PR can be used to create instruction files for liquid-handling robotics or microfluidics platforms to distribute liquids.