Construction of complex and diverse DNA sequences using DNA three-way junctions

Abstract The ability to construct entirely new synthetic DNA sequences de novo is essential to engineering and studying biology. However, the ability to produce long complex synthetic DNA sequences and libraries currently lags behind the ability to sequence and edit DNA 1,2 . All existing DNA-assembly technologies rely on DNA sequence information found within the final construct to direct assembly between DNA molecules 3–11 . As a result of this paradigm, these sequences cannot be extensively optimized specifically for assembly without affecting the final sequence. To fundamentally address this challenge, here we show the development of a new DNA assembly technique named Sidewinder that separates the…

• NS
  National Science Foundation
• CI
  California Institute of Technology
• NI
  National Institutes of Health
• CF
  Center for Environmental Microbial Interactions, California Institute of Technology
• DO
  Division of Biological Infrastructure