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Construction of expression vectors pPICZαA_NS1DENV1–4. (A) Plasmid map: alcohol oxygenase 1 (AOX1) promoter with terminator region. Zeocin resistance gene. pUC ori for start replication in E. coli. Cloning cassette: α-factor secretion signal, NS1 gene, c-myc epitope (not used in this work), His-tag, stop codon and restriction enzyme sites; (B–E) Amino acid sequences that were codon-optimized for yeast expression of viral proteins (DENV1, DENV2, DENV3, and DENV4).
