Here, we demonstrate a new cloning toolkit for the generation of immunological transgenes that is based on Golden Gate cloning. We have termed this toolkit ImPaqT – Immunological Toolkit for PaqCI-based Golden Gate Assembly of Tol2 Transgenes. It includes a range of immunologically focused promoters along with genetic tools and fluorescent proteins that can be assembled using PaqCI-based Golden Gate cloning. Similarly, many groups also use I-SceI insertion approaches to generate transgenic zebrafish (Hoshijima et al., 2016). Although the Golden Gate approach is similarly efficient to other methods to generate transgenesis constructs, the minimal additional sequence and the ability of the Golden Gate method to expand and contract as needed provide flexibility that is beneficial for all of these editing and transgenesis methods.