Systems that allow researchers to precisely control the expression of genes are fundamental to biological research, biotechnology, and synthetic biology. However, few inducible gene expression systems exist that can enable simultaneous, multi-gene control in the important model organism and chassis, Saccharomyces cerevisiae. Here, we repurposed ligand binding domains (LBDs) from mammalian Type I nuclear receptors to establish a family of up to five orthogonal synthetic gene expression systems in yeast. Our systems enable tight, independent, multi-gene control through the addition of inert hormones, and are capable of driving robust gene expression outputs. As a proof-of-principle, we placed expression of four enzymes from the violacein biosynthetic pathway under independent expression control to selectively route pathway flux. Our results establish a modular, versatile, and potentially expandable toolkit for multidimensional control of gene expression in yeast that can be used to construct and control naturally-occurring and synthetic gene networks.

These data were generated to support the development and characterization of a toolkit used to construct orthogonal, small molecule-inducible genetic programs in yeast. The toolkit is based on synthetic transcription factors derived from human type I nuclear receptors fused to zinc finger DNA binding domains and transcription activation domains. In the published work, we extensively characterized synthetic transcription factor performance and orthogonality. As a proof-of-concept, we modulated flux through the violacein biosynthetic pathway by selectively inducing expression of combinations of pathway enzymes. 

Our datasets, comprised of data gathered using flow cytometry and high-performance liquid chromatography, support the four major figures of our manuscript:

1. The mechanism of action, features, and design of synthetic transcription factors for genetic circuit construction in yeast.
2. The characterization of maximum induction, basal expression, and dose curve response for the toolkit elements.
3. The characterization of cross-reactivity and orthogonality among the synthetic transcription factors.
4. The application of the engineered toolkit to the violacein biosynthetic pathway as a proof-of-principle.

The arranged datasets for each figure are included in .xlsx files, named “Fig#_arranged,” and the processed data are presented in .pzfx files, named “Fig#_processed.” They are accompanied by the relevant raw dataset files.

Please refer to the README file for a more detailed description of the data and file structure.

The raw datasets require FlowJo to open, and the processed data files require GraphPad Prism.