Team:Johns Hopkins/Project/PromUTR

Promoters and Terminators

The iGEM registry is lacking in promoter and terminators sequences for yeast, especially when compared with the number available for use in E. coli. In order to address this problem and increase the use of yeast in synthetic biology we have decided to provide a library of promoter and terminator sequences from 12 yeast genes: 4 causing high expression, 4 causing medium expression, and 4 causing low expression. We hope these BioBrick compatible parts will make it easier for future iGEM teams to work with Saccharomyces cerevisiae.

Characterizing the Promoters

In order to provide useful parts with better ease of use, we have decided to not only provide a library of useful yeast promoters to the part registry, but also quantitatively characterize the strength of our promoters by using reference promoters. In a nut shell, we compared the ability of our promoters to express the GFP gene to that of other well characterized promoters in identical laboratory strain. This way, iGEM teams in the future can accurately predict the functionality of our promoters in their strains of interests by referencing the functionality of the reference promoters. A diagram and a brief step-by-step explanation of the process is provided below:

Promoters

Promoter sequences that have been BioBricked:

Terminators

3'UTR sequences that have been BioBricked:

Promoters and 3'UTRs for Golden Gate Assembly

We also have a second promoter/3'UTR library consisting of the same sequences with custom overhangs with BsaI sites rather than the BioBrick sites. These are being used in our two violacein projects, where they will allow Golden Gate assembly of violacein expression cassettes.