pFC30A His6HaloTag T7 Flexi Vector

The pFN29 and pFC20 His₆HaloTag® T7 Flexi® Vectors are used for inducible expression of HaloTag® fusion proteins in E. coli and cell-free systems using the T7 RNA polymerase promoter. Expression levels depend highly on the nature of the protein, but in general the N-terminal HaloTag® fusion protein can increase expression level, enhance refolding and boost solubility of the expressed protein.

The pFN29A/K His₆HaloTag® T7 Flexi® Vectors (Cat.# G8261, G8331) express protein with an N-terminal His₆-HaloTag® dual tag in E. coli T7 cell-free expression systems. The pFC30A/K His₆HaloTag® T7 Flexi® Vectors (Cat.# G8321, G8381) express protein with a C-terminal His₆-HaloTag® dual tag in E. coli T7 cell-free expression systems. Proteins expressed from these vectors can be purified using HaloTag® technology or the polyhistidine tag. The His₆-HaloTag® dual tag enables protein purification with the reusable Ni-resin while retaining the HaloTag® protein covalent labeling properties for binding HaloTag® ligands.

About the Flexi® Vector System

The Flexi® Vector System is a simple, yet powerful, directional cloning method for protein-coding sequences. It is based on two rare-cutting restriction enzymes, SgfI and PmeI, and provides a rapid, efficient and high-fidelity way to transfer protein-coding regions between a variety of Flexi® Vectors without the need to resequence. Directly use recombinant clones and minimize time wasted screening background colonies. The versatile cloning of the Flexi® system means you can choose from a variety of expression systems and fusion tag orientations and then transfer to others as required. Direct transfers can only occur between two N-terminal tagged vectors or from an N-terminal to a C-terminal vector.

Note: Flexi® Vectors contain the lethal barnase gene to reduce background colonies without inserts during the subcloning procedure. Using the Flexi® Vector Cloning System replaces the barnase gene with your insert. These vectors, as purchased, cannot be cultured in normal laboratory strains of E. coli without an insert.