EVOLUTION-GUIDED MULTIPLEXED DNA ASSEMBLY OF DNA PARTS, PATHWAYS AND GENOMES

摘要

The invention relates to a process for assembling DNA parts into multi-kilo base long synthetic DNA constructs. The process generates multiple, synonymous DNA parts in parallel and selects in a combinatorial assembly approach for those sequence variants with the best synthesis and assembly feasibility. DNA parts are sequence optimized and partitioned into synonymous variant designs that serve as redundant building units for higher order DNA assembly. The major stages of the process are: computational partitioning and synonymous recoding of the DNA design, DNA synthesis of sequence variants pools, serial PGR to isolate sets of DNA parts and higher order assembly. As the higher-order assembly does no longer depends on successful synthesis of each DNA part, large-scale DNA designs can be quickly completed allowing for cost-effective and highly parallelised assembly of synthetic bio-designs.