“…Assignment categories were: Bud, Bud Neck, Cell Periphery, Cytosol, ER, Mitochondria, Nuclear Periphery, Nucleolus, Nucleus, Punctate, Vacuole and Vacuole Membrane. (iii) Determination of the SWAT swapping capability (See “Analysis of swapping-procedure efficiency” section); and finally (iv) Sequencing (Anchor-seq) to ensure correct reading frame: We employed a targeted-sequencing strategy detailed in 7 to verify the junction encompassing the 3’ end of the cassette and the 5’ end of each gene. Briefly, we pooled all strains from the SWAT library together, extracted their genomic DNA, sheared it into fragments of 300-800bp that were gel-purified, ligated to Anchor-seq adaptors and submitted to two rounds of PCR.…”