Schematic representation of the Construction of (A) pET30a/His-EDDIE-GFP and (B) pET30a/His-EDDIE-AMPs vectors. (A) pET30a/His-EDDIE-CAD was used to constructed pET30a/His-EDDIE-GFP. This plasmid was derived from pET30a and uses a T7-inducible promoter with lac operator, contains the low-copy pBR322 origin of replication, and encodes the kanamycin resistance gene (kanR) and the lac repressor gene (lacI). The GFP gene was inserted at the Sal I site and replaced CAD gene of pET30a/His-EDDIE-CAD, which at downstream positions of the EDDIE gene, give rise to the vector pET30a/His-EDDIE-GFP for expression of AMPs in E. coli. (B) The AMPs genes were inserted downstream of the carrier protein using overlap primer (arrows) at 168 site. The separate of AMPs and carrier partner between self-cleavage sites while in vitro refolding is shown in the square box, while the self-cleavage site is indicated by an arrow.