Abstract
Successful synthetic biology efforts rely on conceptual and experimental designs in combination with testing of multi-gene constructs. Despite recent progresses, several limitations still hinder the ability to flexibly assemble and collectively share different types of DNA segments. Here, we describe an advanced system for joining DNA fragments from a universal library that automatically maintains open reading frames (ORFs) and does not require linkers, adaptors, sequence homology, amplification or mutation (domestication) of fragments in order to work properly. This system, which is enhanced by a unique buffer formulation, provides unforeseen capabilities for testing, and sharing, complex multi-gene circuitry assembled from different DNA fragments.
Original language | English |
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Article number | 19278 |
Journal | Scientific Reports |
Volume | 6 |
DOIs | |
State | Published - Jan 13 2016 |
Funding
This material is based upon work supported by the Department of Energy, Office of Science, Genomic Science Program (under award number DESC0008834). The authors would like to thank Jen Sheen for providing the plasmid pcoCas9 and Lee E Gunter for critical review and clarifying comments on the manuscript. H.C.D.P. is indebted to CNPQ/FMRP-USP Brazil, Y.Z., M.H.S.G. and J.E.F.F. for his PhD fellowship and previous mentoring. Oak Ridge National Laboratory is managed by UT-Battelle, LLC for the US Department of Energy (under contract number DE-AC05-00OR22725).