Figure 1.

Universality of uLoop. (A) Decoupling of Loop assembly logic. uLoop decouples the propagation elements specific to the organism of choice from the universal assembly logic inbuilt in Loop vectors. (B) Creation of uLoop species-specific vectors from Loop. In uLoop, propagation elements required for transfer and maintenance of plasmids in target organisms are used as modular L0 parts. During the assembly routine, these cassettes are assembled into plasmids alongside with other TUs to generate species-specific plasmids capable of transfer and propagation in the organism of interest. (C) Overhangs and restriction sites used in uLoop/Loop assembly. BsaI overhangs follow the common syntax (8, 13), and SapI overhangs are the same as those described in Loop assembly (10), α(ATG), β(GCA), γ(TAC), ε(CAG) and ω(GGT). (D) Source of uLoop vector kits. Vectors from the synthetic biology community and the pCAMBIA vector were domesticated for BsaI and SapI and ‘minimized’ by removing elements not related to basic plasmid function. Two antibiotic resistance versions were generated and then the Loop schema was incorporated to generate odd and even plasmids for each version of vector kits.