CIDAR MoClo: Improved MoClo Assembly Standard and New E. coli Part Library Enable Rapid Combinatorial Design for Synthetic and Traditional Biology
journal contributionposted on 15.01.2016, 00:00 authored by Sonya V. Iverson, Traci L. Haddock, Jacob Beal, Douglas M. Densmore
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.
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Traditional BiologyMultipartNew EGolden Gate assembly methodsEscherichia colibinding sitesMoClo protocolsassembly standardsMoClo Assembly StandardDNA part librariesCIDAR MoCloMoClo LibraryOptimized protocolsMoClo assembly frameworkreaction timeDNA partscombinatorial designcontrol plasmidsDesign Automation Research labcoli Part Library Enable Rapid Combinatorial Designcoding sequence