Figure 7. Combinations of up to eleven built from sub-cloned donor fragments.

Sub-cloned donor fragments of 2–5 cassette combinations (excised with the core 2 enzymes) were inserted into both 3 and 6 cassette recipient plasmids, successfully creating new combinations of 5, 6, 7, 8 (two combinations thereof), 9, 10 and 11. Insertions were confirmed by PCR screening using primers that flanked the MCS region. Each insertion added ∼270 bp to the size of the previous recipient plasmid.