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pFC30K His6HaloTag T7 Flexi Vector

by Promega
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  • Overview
  • Protocols
  • Specifications
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  • 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.

    pFN29A His6HaloTag® T7 Flexi® Vector GenBank® Accession Number JN874648 and vector sequence text file.

    pFC30A His6HaloTag® T7 Flexi® Vector GenBank® Accession Number JN874649 and vector sequence text file.

    pFN29K His6HaloTag® T7 Flexi® Vector GenBank® Accession Number JN874650 and vector sequence text file.

    pFC30K His6HaloTag® T7 Flexi® Vector GenBank® Accession Number JN874651 and vector sequence text file.

  • Protocols

    Complete Protocol

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    pFN29A His6HaloTag® T7 Flexi® Vector Product Information

    PDF (167 KB)

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    pFN29K His6HaloTag® T7 Flexi® Vector Product Information

    PDF (166 KB)

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    pFC30A His6HaloTag® T7 Flexi® Vector Product Information

    PDF (198 KB)

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    pFC30K His6HaloTag® T7 Flexi® Vector Product Information

    PDF (166 KB)

  • Certificate of Analysis

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    Storage Conditions

    -30C TO -10C

    Use Restrictions

    For Research Use Only. Not for Use in Diagnostic Procedures.

  • Resources

    Software and Firmware