Chromatin is the nucleo-protein complex that organizes genomic DNA in the nuclei of eukaryotic cells. Transcriptional regulators and enzymes are key mediators of chromatin states that affect gene expression and consequently cell phenotype. Many natural chromatin mediators contain sub-domains that can be isolated and recombined to build novel regulators and live cell probes. A wide variety of synthetic chromatin proteins can be constructed from the diverse array of known natural chromatin proteins and synthetic variants. The process of engineering chromatin mediators and probes produces new tools for cell engineering and deepens our understanding of the mechanism by which chromatin features, such as modifications of histones and DNA, contribute to the epigenetic states that govern DNA-templated processes. To support efficient exploration of the large combinatorial design space of synthetic chromatin proteins, we have developed a Golden Gate assembly method for one-step construction of recombinant protein-encoding DNA. A set of standard 2-amino acid linkers allow facile assembly of any combination of up to four protein modules, obviating the need to design different compatible overhangs to ligate different modules. Beginning with the identification of protein modules of interest, a synthetic chromatin protein can be built and expressed in vitro or in cells in under two weeks.