We expect CAN to find use as a tool in CPS for efficiently ligating peptides suffering from sluggish thioesters and/or poor solubility. Interestingly, since lower-concentration CAN is expected to maintain its ligation rate, the relative benefit of CAN over solution NCL will increase at further reduced peptide concentrations (e.g., compared to NCL at 500 nM, STEVE/KENT CAN is predicted to be ~12,000,000-fold faster). The current CAN methodology is expected to be applicable to many CPS strategies, especially since the reaction rate enhancement is relatively insensitive to linker attachment points. For example, 64% of all possible Cys and Ala ligation junctions in the E. coli 50S and 30S ribosomal proteins are CAN-compatible (Lys within 20 AA's of the junction on both the N-and C-terminal peptides). CAN will also be particularly useful for sluggish ligations using unnaturally thiolated amino acids that enable NCL outside of Cys, such as thiolated valine. Furthermore, CAN should be compatible with other peptide linker strategies, due to the simplicity of preparing both azide and DBCO peptides.