In order to achieve maximum expression in E. coli system, Jcat (Grote et al. 2005) was used for reverse translation and optimization of codons. Jcat is a rapid and easy method that could ensure maximum expression by calculating the GC content and CAI value (Grote et al. 2005). Restriction sites of BamHI and KpnI were added to the reverse translated sequence. The optimized sequence was then cloned into pET30a plasmid using Snapgene software.

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