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Universal regulatory library for any cell type

Periodic Reporting for period 1 - UNILIB (Universal regulatory library for any cell type)

Reporting period: 2019-07-01 to 2020-12-31

Problem being addressed:

The lack of a reliable promoter resource has hindered the development of many synthetic biology applications, especially based on Eukaryotic chassis.

Importance for society:

Many application that are based on yeast can be made possible, including but not limited to various foot products, biologics, and other novel engineered living materials.

General objective:

To develop a promoter resource in yeast that can function reliably in any growth condition.
We designed an oligo library encoding 200,000 variants containing up to 3 binding sites from a list of 42 TF binding sites identified in MRG-Grammar. The library was cloned upstream to a minimal yeast synthetic promoter and characterized via SORT-Seq. The library generated a spectrum of expression levels achieving the resource goal.
We now possess a promoter resource in yeast that heretofore did not exist. Prior to publication we expect to characterize the library in another 4-5 growth conditions, thus completing the resource. We expect the library to be widely impactful on yeast based applications.