The Jamaican Click Beetle (Pyrophorus plagiophthalamus) has dorsal and ventral bioluminescent organs with the unique ability to express multiple colors: green, yellow-green, yellow, and orange. The specific colour emitted is determined by the sequence of the luciferase protein. In their natural population, alleles coded to produce orange are increasing in frequency, suggesting there is a selective pressure on orange colouration. The mechanism of a trait’s evolution favouring one allele over another is at the heart of evolutionary biology. Adaptation occurs through the accumulation of rare mutations in a population, with selection selecting individuals with advantageous survival traits. The objective of this study is to create a dynamic model via the expression of synthetic alleles in bacteria to investigate the relationship between mutations in the luciferase gene and colour shifts. By exploring the realm of genetic variation, the aim is to determine if the increased prevalence of orange alleles in Jamaica signifies an adaptive trait, as previous literature suggests, or simply the most common mutational pathway. Synthetic luciferase plasmids of the natural alleles have been expressed, and a primary experiment created a single mutation to the ventral yellow-green allele which led to a green hue, a shift in the direction away from orange. From these initial expressions of synthetic genes, a single-site variation library (SSVL) was ordered to fully explore the space of change in the family of Jamaican click beetle luciferase. Exploring the space of change in click beetle luciferase unveils a novel approach to studying the functionality of luciferase proteins, an important tool in bioluminescence assays and various research methodologies.