BBF RFC #111: Inducible expression vector with His tag fusion for purification of RFC25 parts

Abstract

This Request For Comments (RFC) describes a new expression vector based on the pET expression system and designed for BioBrick RFC25 parts. This design includes all the features of pET expression vectors with a new origin of replication, resistance marker and Multiple Cloning Site (MCS). This MCS contains new restriction sites and a 5’ poly- histidine tag that allows the purification of expressed RFC25 parts. Previous attempts have been prevented by restriction site incompatibility, but using RFC25 parts and their restriction sites enables 5’ fusion to these parts. This new expression vector enables the cloning of RFC25 parts with a 5’ His tag in a single step. This allows the inducible expression of these parts and the easy purification of protein products.