Fig. 1

HTP gene synthesis platform used to produce 4992 synthetic genes encoding venom peptides. This pipeline includes 7 steps that allow the successful synthesis of multiples of 96 genes. The first step corresponds to gene design and codon optimization; from multiple peptide sequences, DNA sequences are designed and optimized for expression in E. coli, using the ATGenium codon optimization algorithm. In steps 2, 3 and 4 oligonucleotides required for gene assembly are designed using the NZYOligo designer, synthesised and assembled by PCR using optimal conditions, respectively. Synthetic genes are cloned using NZYTech LIC protocol into the E. coli expression vector pHTP4. Bacterial transformation and DNA preparations are accomplished using high-throughput protocols. DNA sequences are checked for the presence of sequence errors using a high-throughput sequencing analysis tool. All steps are automated using a Tecan liquid handling system. The plate containing the 96 clones (step 8) is ready to go through the peptide production pipeline