Fig. 2
From: Simplified plasmid cloning with a universal MCS design and bacterial in vivo assembly
Principle of the UMCS and corresponding primer design. a The UMCS sequence used in this study contains two homologous linker regions that are connected by the SalI recognition sequence. The flanking HindIII and XbaI recognition sequences were designed for replacing the original MCS. All three recognition sites can be freely substituted for other sequences. b The UMCS was first synthesized as two ssDNAs (each with a length of 42 bp). After annealing and ligating, the UMCS was incorporated into the vector, thus replacing the original MCS. c The recommended primer design to maximize assembly efficiency. The homologous linker region (12 ~ 15 bp) is included in the 5′-terminus of each primer, and the Tm of the template binding region was designed to be approximately 55 °C