Figure 8.
Strategies for DNA assembly across scales relevant for plant biosystems design. (a) Major platforms and key factors considered for executing large DNA assemblies across scales. (b) Detail of the CCTL method. crRNA sequence is from [123]; overhang is underlined with the last 4 nt being programmable [124]. ¥Produce scarless assemblies (NEBuilder allows ssDNA oligos in substitution of homologous overlap).¤May leave scars. Undesired type IIS restriction sites can be partially masked by oligos [122]. For BRAID systems, see [120]. §Hybrid system using golden-gate followed by in vivo homology-based assembly [129]. CX: complex design and execution; PL20: parts size/length beyond 20 kb; RS: repetitive sequences; SS: secondary structures; IRS: internal restriction sites. Recombination-based approaches (e.g., Gateway) were omitted due to limited use for biosystems design.