lexAop (LexAop2) regulatory sequences are fused upstream of a removable cassette comprised of FLPD5 and Hsp70b and SV40 3' UTRs surrounded by RSRT recombination sites. Downstream of the removable cassette is a coding sequence encoding TEVp plus a transmembrane sequence (Tag:M(rCd2)) that targets TEVp to the cell surface (note that when this fusion protein is expressed in cell culture, the extracellular TEVp protein shows only limited protease activity). The N-terminus of the coding sequence has the C. elegans PAT3 signal peptide sequence for endoplasmic reticulum targeting, while the C-terminus has the Kir2.1 ER export signal to improve trafficking to the plasma membrane, followed by the P10 3' UTR. Tag:MYC has been introduced in between the TEVp and Tag:M(rCd2) sequence. Removal of the cassette allows the TEVp-Tag:M(rCd2) protein to be expressed under control of lexAop. (GenBank MN658787.) Used in the first (release) step of the BacTrace system.