Many of our basic parts (promoters, terminators, and fluorescent proteins) are courtesy of the Dueber Lab at UC Berkeley, our host this year. Our iGEM team chose the targeting proteins and sequences from the the Yeast GFP Database created by the O'Shea and Weissman Labs at UCSF, and either PCRed them from genomic DNA or synthesized them via gene synthesis ourselves. We then combined these parts with the basic parts provided by the Dueber Lab to build all the devices we utilized.
All the higher-order (cassette, multigene, full MiCode) device cloning, which mainly involved Golden Gate Assembly, was done by our iGEM team. While some of the Golden Gate backbones were provided by the Dueber Lab, many of the parts specific to MiCode project, specifically the backbones for multigene and full MiCode assembly, were designed and cloned by us. Additionally, we produced all the yeast strains necessary throughout our entire iGEM project and did all the image acquisition and analysis.
The leucine zipper sequences were provided courtesy of the Keating Lab at MIT, in a collaborative effort to identify an orthogonal interaction set. With the sequences provided, we constructed the leucine zippers via gene synthesis, combined them into our MiCode construction scheme, and assayed them in yeast.
On the software side, we wrote the MATLAB code, constructed the Cell Profiler pipeline for automated image analysis, and tested it using images generated by our iGEM team. Matlab object identification functions from the community forum were used as templates for the basis of our own code. We developed CellProfiler pipelines with the aid of the CellProfiler manual and sample pipelines from the online forum.
Website, Presentation, and Poster
For the presentation and poster, any images not of our creation are cited where relevant.