Recombinant macromolecular complexes are often produced using the baculovirus system with multigene expression vectors [1]. However, multigene vectors for the baculovirus system are traditionally built using complex processes that are time-consuming and require substantial skill. To streamline the process of baculovirus vector building, we developed the MoClo Baculo toolkit [2]. MoClo Baculo simplifies the construction of multigene expression vectors for the baculovirus system. MoClo Baculo is also compatible with the MoClo Yeast toolkit [3], therefore enabling protein expression using both the baculovirus system and yeast. The MoClo toolkit does not require PCR, primers, or sequencing of intermediate products, significantly reducing the time and effort involved in vector construction. Based on Golden Gate assembly, MoClo Baculo minimises mutations and therefore allows for quick validation of intermediate plasmids through restriction digestion. The system provides end-to-end modular cloning, enabling easy exchange of tags, open reading frames, and even promoters and backbone when switching between baculovirus, yeast, and bacterial expression systems. As a proof of principle, we successfully used MoClo Baculo to construct baculovirus and yeast multigene vectors expressing the four-, five- and six-subunit human Polycomb repressive complex 2. Overall, the MoClo Baculo toolkit streamlines and accelerates the construction of multigene expression vectors for the baculovirus system and provides compatibility with yeast as an alternative expression system.