The files provide details of the genetic construct designs, with a focus on obtaining the Drosophila melanogaster morphogens Hedgehog, Decapentaplegic, and Wingless. The experiments were performed at the Center for Medicinal Chemistry (CQMED), which applies SGC protein purification strategies for drug-discovery approaches. These strategies involve generating different constructs of aim proteins and testing a small-scale high-throughput assay to identify the best constructs for generating large-scale aim proteins. The Center for Medicinal Chemistry (CQMED), which is part of the SGC, is where the experiments were performed. This work involved four variants of each morphogen protein, with the following variables: full length; truncated at the N-terminal; truncated at the C-terminal; or both truncated. Each of these four variants was cloned into five different plasmids using Ligation Independent Cloning (LIC): pNic28-Bsa4 (adds a His-tag to the N-terminal region, linked by a TEV cleavage site), pGTVL2 (adds a His-tag and a GST tag to the N-terminal region, linked by a TEV cleavage site), pNIC-CTH0 (adds a His-tag to the C-terminal region, linked by a TEV cleavage site), pNIC-Zb (adds a His-tag and a Z-basic tag (ZB) to the N-terminal region, linked by a TEV cleavage site), and pSUMO-LIC (adds a His-tag and a SUMO protein). N-terminal, linked by a SUMO cleavage site). All of these vectors have a SacB gene for negative selection in media containing 5% sucrose, flanked by a BsaI restriction site, specific LIC sequences and a promoter to induce protein expression with IPTG. The optimising design of the primers with the planning plates for 60 constructs (3 morphogens x 4 variants x 5 vectors) was performed using the “LIC construct and primer design tool” program (https://gbarbosabio.shinyapps.io/CQMED_Primer_Construct_/) developed by Professor Guilherme during an internship at CQMED.