A systematic study of building-block compatibility with DNA-templated macrocycle synthesis. Using macrocycle 8a as a benchmark of “normal” reactivity, we selected a set of six distinct amino acids for substitution, one at a time, into the macrocycle at each of the three building block positions (R1, R2, and R3). Macrocycle series 11 (substitution at R1), 12 (substitution at R2), and 13 (substitution at R3) were synthesized using the corresponding reagent sets and starting materials 1a, 5a, and 6a, respectively. Percent yields of each macrocycle as analyzed by PAGE using internal standards are shown in the table.