Plant-derived medicines are prone to shortages because their agriculture-based supply chains are vulnerable to geopolitical and environmental factors. Recreating the biosynthesis of these medicines in a scalable organism like Baker’s yeast would provide an economical alternative to plant cultivation that is resilient to regional and global disruptions and scalable to meet global demands. This application note showcases the engineering of Baker’s yeast to synthesize the nightshade-derived tropane alkaloids hyoscyamine and scopolamine. Whole-cell engineering leveraged Twist Gene Fragments for rapid screening and integration of pathway elements into yeast, removing a critical bottleneck in the engineering workflow.