Engineered Fluorescent Sec61β Constructs

mEmerald-Sec61β and mCherry-Sec61β were acquired from Addgene (plasmids 54249 and 49155, respectively). GFP-KDEL was cloned as previously described (Merta et al., 2021 (link)). In brief, a pDsRed2-ER vector (632409; Clontech) was digested with AgeI and HindIII to remove DsRed2, and a PCR-amplified GFP sequence with AgeI and HindIII sites was ligated into the vector. SNAP-Sec61β and Halo-Sec61β were cloned as previously described (Bottanelli et al., 2016 (link)). Briefly, mEmerald-Sec61β (above) was digested with NheI and BgIII to remove mEmerald, and a PCR-amplified HaloTag or SNAP-tag sequence with NheI and BgIII sites was ligated in mEmerald’s place.