Construction of pET-based YbeY overexpression plasmid
The pET-ybeY overexpression plasmid was constructed using the CPEC (Circular Polymerase Extension Cloning) method (Quan and Tian, 2011). The primers were designed to amplify the pET-Orange plasmid backbone and the coding region of the ybeY gene from the E. coli genome (sequences are detailed in Table S3). Phusion™ HF DNA Polymerase (Thermo Scientific, #F530L) was used following the manufacturer protocol.