Fig. 3.

Efficiency of Golden Gate assembly of different APC/C constructs containing 10–15 expression cassettes. To test the efficiency of the GoldenBac system, Golden Gate assemblies were performed using different numbers of expression cassettes. Ten clones resulting from each assembly reaction were picked, digested with EcoRV, and analyzed via agarose gel electrophoresis. A schematic of the DNA sizing ladder and the predicted band pattern for each assembly reaction is shown to the left of the respective agarose gel. Clones demonstrating correct assembly based on the pattern of bands are marked with a green asterisk. The reaction maintains 80% efficiency with assembly of up to 14 expression cassettes. One of the clones containing 14 genes was then used for baculovirus generation and recombinant protein production of the APC/C complex