We submitted two parts to the iGEM registry: an N-carbobenzyloxy (CBZ) cleavage enzyme part, and a composite part consisting of a promoter, the CBZ cleavage enzyme, and a terminator.
This basic part contains the coding sequence for the N-carbobenzyloxy (CBZ)
cleavage enzyme necessary to remove the CBZ protecting group from the
N-terminus of amino acids. The coding sequence was derived from Nanduri
Nanduri, V.B., Goldberg, S., Johnston, R., and Patel, R.N. (2004). Cloning and expression of a novel enantioselective N-carbobenzyloxy-cleaving enzyme. Enzyme and Microbial Technology 34, 304–312. , and we included an RBS sequence upstream of the coding sequence.
This composite part contains a promoter (BBa_J23118), RBS, the CBZ cleavage enzyme, and a terminator (BBa_J61048) to make a fully functional BioBrick. In our biocontainment system, this BioBrick cleaves CBZ from N-CBZ-leucyl-tRNA to produce wild-type leucyl-tRNA. This part can be used by other iGEM teams to selectively cleave the CBZ protecting group from amino or hydroxyl groups in organic reactions.