Ribosome protein sequences were processed in Aligator with default thioesters (I, L, K, T, V accepted; A, C, F, H, G, M, R, S, W, Y preferred), an expanded set of thiols (C, A, and V), and a 60-aa segment limit to obtain a list of up to 1000 top strategies each. The resulting "<Protein Name> All Strategies.txt" files were processed in BracketMaker via our Aligator Import tool to obtain the best-scoring ligation order for each set of segments, giving a list of up to 1000 top brackets per protein in bracket notation (the strategies for IF2 were too large for Auto-Bracket to process [22 segments] and resulted in memory errors, so we divided these strategies into smaller groups of segments separated by two available C sites [which must be used last in either order] and ran Auto-Bracket separately, then manually combined the top brackets from each to obtain the top full IF2 bracket). The following Auto-Bracket settings were used, representing available hydrazide-NCL chemistry commonly used in our lab: One-Pot Desulfurization = FALSE, One-Pot Cys Deprotection = FALSE, Cys Protecting Group = "Acm", 5