CIDAR MoClo: Improved MoClo Assembly Standard and New E. coli Part Library Enable Rapid Combinatorial Design for Synthetic and Traditional Biology

Multipart and modular DNA part libraries
and assembly standards
have become common tools in synthetic biology since the publication
of the Gibson and Golden Gate assembly methods, yet no multipart modular
library exists for use in bacterial systems. Building upon the existing
MoClo assembly framework, we have developed a publicly available collection
of modular DNA parts and enhanced MoClo protocols to enable rapid
one-pot, multipart assembly, combinatorial design, and expression
tuning in Escherichia coli. The Cross-disciplinary
Integration of Design Automation Research lab (CIDAR) MoClo Library
is openly available and contains promoters, ribosomal binding sites,
coding sequence, terminators, vectors, and a set of fluorescent control
plasmids. Optimized protocols reduce reaction time and cost by >80%
from that of previously published protocols.