Part:BBa_K1352002

Abstract

We describe the creation of Biobrick constrct Bba_K1352002 which expresses autotransporter Ag43 with an inserted FLAG epitope tag at codon 148, flanked by multiple cloning restriction sites. Two 17 amino acids long beta hairpins were removed from Ag43 in order to eliminate native aggregation properties. We describe the design and construction of the new Biobrick, and its verification using restriction digestion and DNA sequencing. We show that removal of β-hairpins removes auto-aggregation capability of Ag43, creating a useful reagent that can be used for E. coli surface display.