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Luciferases screened to identify orthogonal probes. Mutant libraries were prepared using site-saturation mutagenesis. Library sizes varied based on the degree of saturation enforced at each residue. Such determinations were made based on sequence conservation among the insect luciferase family and residue location. Targeted residues are colored, and most lie in close proximity to the bound luciferin (PDB: 4G36).
