*2.1. DNA Constructs*

The HIV consensus clade C envelope, Influenza HA1, and Zika prME DNA vaccines used in these studies are as previously described [39–41]. Figures 1A, 4A, and 5A have been adapted from figures from these studies.

The sequences for murine IL-36 alpha, beta, and gamma were obtained from Uniprot (Q9JLA2-1, Q9D6Z6-1, Q8R460-1). These sequences have been modified to be RNA and codon-optimized in order to exploit the host's natural codon preference and enhance protein expression. Furthermore, a highly efficient IgE leader sequence was inserted at the 5' end of the IL-36 gene to promote efficient secretion of the protein. These full-length optimized IL-36 cytokine plasmids are known henceforth as opt-36α, opt-36β, and opt-36γ.

Recent work by Towne et al. has demonstrated the need for truncation of IL-36 cytokines nine amino acids N-terminal to a conserved *A*-*X*-*Asp* motif, for full activity [42]. The second set of IL-36 plasmids have been truncated according to the data presented in the paper and are henceforth known as opt-36αt, opt-36βt, and opt-36γt. All inserts were modified as previously explained above for enhanced expression and cloned into the pGX0001 backbone (Genscript, Piscataway, NJ, USA) [43].
