Figure 1.

One-pot GGA of 52 fragments into a destination vector. (A) Schematic of the 52-fragment lac operon cassette assembly. Assembly inserts were generated by PCR amplification and assembled into a destination vector containing an antibiotic resistance marker. (B) Example outgrowth plate used for colorimetric scoring by a reverse blue-white screen. Correctly assembled 52 insert constructs form blue colonies upon cellular transformation, and incorrectly assembled constructs produce white colonies.