Figure 5

Strategy to combine different repetitive sequences in a modular fashion. Antiparallel oligonucleotide pairs with different repetitive sequences (orange and blue) can be fused on one plasmid if their Type IIS recognition sites (IIS) are designed in a way that they create compatible overhangs. Moreover, our strategy is not restricted to specific endonucleases. The use of any combination of one Type IIP (IIP) and two different compatible Type IIS enzymes is possible. Upon insertion of the first annealed and digested oligonucleotide pair (orange) via IIS and IIP, the resulting vector can be digested with IIS' and IIP to allow the incorporation of a second double-stranded oligonucleotide pair (blue).