Figure 3. MCS functionality in the recipient plasmid.

The recipient plasmid modified by insertion of the designed MCS was digested individually with Age I, Spe I, Nhe I, Bsr GI, Mlu I, Asc I, Pac I, Acl I, and Dra III to confirm their usability. All digestions linearized the plasmid, except for Spe I and Mlu I that were also present in the plasmid backbone and thus gave an expected two fragments (this did not impact on their role in the cloning strategy).