Blue Gate Assembly™ — SapI

<p><em>Type IIS restriction enzymes are unique endonucleases that produce neat and defined overhangs from cleavage sites that occur outside of the enzyme binding site. Type IIS restriction enzymes are powerful tools in biotechnology because they are commonly used for specialized molecular cloning like de novo genetic construct assembly and building DNA variant coding libraries. The power that this class of enzyme holds is the ability to design and implement modular cloning strategies for inserting a variable coding region in an ORF for investigating the function of a specific residue in a protein. The production of specifically defined 3 nt overhangs instead of donor DNA with long homology arms for cloning allows site-directed mutagenesis for building variant coding sequences. This article will provide a brief review of Type IIS endonuclease use in biotechnology and an example of how this technology can be used for developing a variant coding sequence for the engineering of Psp polymerase for improved fidelity.</em></p> <p><a href="https://medium.com/@radegenbio/blue-gate-assembly-sapi-393a71a8b155"><strong>Website</strong></a></p>