Tetrahymena thermophila is a ciliate single-celled eukaryote that is extensively studied as a model organism, particularly in the area of chromatin remodeling and DNA repair, where key discoveries have elucidated nucleosome structure and telomerase function. Fluorescence tagging with GFP and RFP is a common way to study protein interactions in T. thermophila and typically makes use of recombination of the fusion gene product into the RPL29 locus (exogenous tagging system). GFP/RFP localization along with immunoprecipitation and gene expression studies can elucidate unknown interactions and uncover pathways previously uncharacterized. This project was to develop constructs that are directed to a beta tubulin 1 locus and exogenously express fluorescent-tagged genes (GFP and RFP), while maintaining compatibility with Gateway cloning technology. This will give the ability to visualize protein dynamics while inducing protein expression in live cells, easily moving genes into the plasmid and examining co-localization of tagged genes expressed in different loci. Our plasmids contain a metallothionein (MTT-1) inducible promoter and fluorescence tags of GFP or RFP. Through homologous recombination, genes will be inserted into the beta tubulin 1-1 (btu1-1) locus, and following transformation cells are selected by paclitaxel resistance. Of particular interest to this work are proteins involved in DNA repair, in which studying sirtuin and NER-related homologues and making comparisons with previous data provide a benchmark to judge the effectiveness of our plasmid construction.