Figure 1
MK system entry cloning strategy. (A) Overview of required primer components for amplifying a new DNA part (A-B example). In addition to part homology, each primer contains a BpiI restriction site for directional LI cloning, and a BsaI restriction site for LII cloning. (B) Performing a BpiI cut-ligation with the amplified part and the universal p641-BpiI backbone generates a LI entry clone for the MK system. The p641-Bpi plasmid contains T1 and T0 terminator sequences flanking the cloned insert, a gentamicin resistance cassette, and ccdB negative selection.
