Sustainable production of chemicals, materials, and pharmaceuticals is increasingly performed by genetically engineered cell factories. Engineering of complex metabolic routes or cell behavior control systems requires robust and predictable gene expression tools. In this challenging task, orthogonality is a fundamental prerequisite for such tools. In this study, we developed and characterized in depth a comprehensive gene expression toolkit that allows accurate control of gene expression in Saccharomyces cerevisiae without marked interference with native cellular regulation. The toolkit comprises a set of transcription factors, designed to function as synthetic activators or repressors, and transcription-factor-dependent promoters, which together provide a broad expression range surpassing, at high end, the strongest native promoters. Modularity of the developed tools is demonstrated by establishing a novel bistable genetic circuit with robust performance to control a heterologous metabolic pathway and enabling on-demand switching between two alternative metabolic branches.

中文翻译：

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酿酒酵母复杂遗传电路工程的综合工具包

基因工程细胞工厂越来越多地实现化学，材料和药物的可持续生产。复杂的代谢途径或细胞行为控制系统的工程设计需要强大且可预测的基因表达工具。在这项艰巨的任务中，正交性是此类工具的基本前提。在这项研究中，我们开发并深入表征了一个全面的基因表达工具包，该工具包可精确控制酿酒酵母中的基因表达。对天然细胞调节无明显干扰。该工具包包括一组转录因子，设计用作合成激活剂或阻遏物，以及转录因子依赖性启动子，它们共同提供了一个广泛的表达范围，在高端范围内超过了最强的天然启动子。通过建立具有强大性能的新型双稳态遗传电路来控制异源代谢途径并实现两个替代代谢分支之间的按需切换，可以证明所开发工具的模块化。