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Overview of Yeast Toolkit
Combinatorial yeast expression cassette library
We are creating a library of yeast promoters and 3'UTRs that are fully compatible with the BioBrick standard. The sequences come from the upstream and downstream region of 12 yeast genes, and are associated with varying levels of expression strength: 4 for strong expression, 4 for medium expression, and 4 for weak expression.
In addition to our BioBrick compatible promoters and 3'UTRs, we have also given the same sequences designed overhangs with BsaI sites to facilitate Golden Gate assembly, which we are using in our violacein projectviolacein project. The Golden Gate assembly method has advantages over BioBrick assembly in that multiple pieces can be ligated together at once with no scar.
Veast shuttle vector library
[ DANIEL BIBL SHOULD WRITE A PARAGRAPH OR SO ABOUT WHAT WE DID (not why we did it, that's background) HERE]
Violacein pigment: a yeast reporter
Violacein purple pigment production serves as a simple way to test a multiple gene pathway (such as vitamin production) in an expression vector or neochromosome. If the gene construct is transformed and the organism becomes purple, the system is successful. As our main project is in yeast and since purple bacteria was achieved by Cambridge iGEM 2009, we focused on producing violacein in the eukaryotic S. cerivisiae yeast. We chose violacein specifically as a Boeke Lab rotation student used the Build-A-Genome method to engineer and sequence violacein building blocks for yeast. In essence, we had a head start with the violacein pathway as a significant portion of the gene assembly has been completed.
