MTK Primer Generator

This tool outputs a list of primers that can be used to "domesticate" an input sequence for the Mammalian ToolKit (MTK), a modular and extendable Golden-Gate-based cloning toolkit. For a given input sequence, the output below describes PCRs that can be used to add particular restriction sites to the ends of the sequence and remove internal restriction sites (using a silent mutation if applicable) to make the sequence compatible with the MTK system. The gel-isolated PCR products can be combined with the part-entry plasmid (MTK0-027) via a Golden-Gate reaction to create a new part plasmid of the desired type. 

The suggested primers may be slightly longer than ideal if there is homology between the adapter sequence at the beginning of the primer and the plasmid itself, so I recommend checking primer specificity and melting temperatures using an application like Benchling and adjusting primer length if necessary.

See the publication here for more details about the MTK system.

You can also find a command line version of this program on GitHub here.