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HQ and GST Tag Flexi® Vectors for E. coli and Cell-Free Protein Expression

The vectors below are used for inducible expression of HQ- and GST-tagged fusion proteins in E. coli and cell-free systems using the T7 RNA polymerase promoter. The HQ tag and polyhistidine tag (His) are comparable in their affinity for Ni ions and will bind to all His-binding surfaces and resins. In certain cases the HQ-tagged proteins can be eluted from the affinity columns at lower concentrations of imidazole—a property useful for some downstream applications such as enzymatic reactions. As with His tag, proteins can be expressed from bacterial, insect and mammalian systems and purified under either native or...

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The Flexi® Vector System is a simple, 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. Direct transfers can only occur between two N-terminal tagged vectors or from an N-terminal to a C-terminal vector.

pFN2A/K (GST) Flexi® Vectors are designed for protein expression with an N-terminal GST tag in E. coli and T7 cell-free expression systems.
pFN6A/K (HQ) Flexi® Vectors are designed for protein expression with an N-terminal HQ tag in E. coli and T7 cell-free expression systems.
pFC7A/K (HQ) Flexi® Vectors are designed for protein expression with an C-terminal HQ in E. coli and T7 cell-free expression systems.
pF1A/K T7 Flexi® Vectors (Cat.# C8441, C8451) are designed for inducible expression of native untagged protein.

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.

Features - Benefits

Easy to Implement and Reliable: Choose between traditional His-affinity and GST-affinity resins for standard protein purification and prokaryotic expression applications.
Cost-Effective: Technology for reusable and cost-efficient Ni (His-affinity) and gluthathione (GST-affinity) resins.
Versatile Cloning: Choose from a variety of expression systems and fusion tag orientations and then transfer to others as required (for Flexi® system).
Time Savings: Barnase insert (Flexi® system) decreases the number of background colonies, allowing efficient transfer of genetic constructs.

Applications

Bacterial and cell-free protein expression and purification with GST tag technology.
Bacterial and cell-free protein expression and purification with HQ (His) tag technology.

For more information, see the Protocols & Applications Guide.

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Products
Specifications
Figures & Tables
Product Resources
Patents & Disclaimers

Prices valid for customers of Promega AG only

Product	Size	Conc.	Catalog #	*List Price	Add to Cart
pFN2A (GST) Flexi® Vector Protocol MSDS Components Close Window pFN2A (GST) Flexi® Vector Components pFN2A (GST) Flexi® Vector C846A1 x 20μg	20μg	-	C8461	CHF 476.00	Add to cart
pFN2K (GST) Flexi® Vector Protocol MSDS Components Close Window pFN2K (GST) Flexi® Vector Components pFN2K (GST) Flexi® Vector C847A1 x 20μg	20μg	-	C8471	CHF 476.00	Add to cart
pFN6A (HQ) Flexi® Vector Protocol MSDS Components Close Window pFN6A (HQ) Flexi® Vector Components pFN6A (HQ) Flexi® Vector C851A1 x 20μg	20μg	-	C8511	CHF 476.00	Add to cart
pFN6K (HQ) Flexi® Vector Protocol MSDS Components Close Window pFN6K (HQ) Flexi® Vector Components pFN6K (HQ) Flexi® Vector C852A1 x 20μg	20μg	-	C8521	CHF 476.00	Add to cart
pFC7A (HQ) Flexi® Vector Protocol MSDS Components Close Window pFC7A (HQ) Flexi® Vector Components pFC7A (HQ) Flexi® Vector C853A1 x 20μg	20μg	-	C8531	CHF 476.00	Add to cart
pFC7K (HQ) Flexi® Vector Protocol MSDS Components Close Window pFC7K (HQ) Flexi® Vector Components pFC7K (HQ) Flexi® Vector C854A1 x 20μg	20μg	-	C8541	CHF 476.00	Add to cart
pF1A T7 Flexi® Vector Protocol MSDS Components Close Window pF1A T7 Flexi® Vector Components pF1A T7 Flexi® Vector C844A1 x 20μg	20μg	-	C8441	CHF 424.00	Add to cart
pF1K T7 Flexi® Vector Protocol MSDS Components Close Window pF1K T7 Flexi® Vector Components pF1K T7 Flexi® Vector C845A1 x 20μg	20μg	-	C8451	CHF 424.00	Add to cart

You Also May Be Interested In

GenBank^®/EMBL Accession Number

pFN2A (GST) Accession Number AY753578; pFN2A (GST) Vector sequence text file.
pFN2K (GST) Accession Number AY753579; pFN2K (GST) Vector sequence text file.
pFN6A (HQ) Accession Number, DQ132629; pFN6A (HQ) Vector sequence text file.
pFN6K (HQ) Accession Number DQ132630; pFN6K (HQ) Vector sequence text file.
pFC7A (HQ) Accession Number DQ133906; pFC7A (HQ) Vector sequence text file.
pFC7K (HQ) Accession Number DQ133907; pFC7K (HQ) Vector sequence text file.
pF1A T7 Accession Number AY753576; pF1A T7 Vector sequence text file.
pF1K T7 Accession Number AY753577; pF1K T7 Vector sequence text file.

Storage Conditions

Store vectors at –20°C.

For product intended use please see Patents & Disclaimers tab.

Figures

Figure 1. pFN2A (GST) Flexi® Vector.

Figure 2. pFN2K (GST) Flexi® Vector.

Figure 3. pFN6A (HQ) Flexi® Vector.

Figure 4. pFN6K (HQ) Flexi® Vector.

Figure 5. pFC7A (HQ) Flexi® Vector.

Figure 6. pFC7K (HQ) Flexi® Vector.

Figure 7. pF1A T7 Flexi® Vector.

Figure 8. pF1K T7 Flexi® Vector.

Related Citations
MDM2 protein-mediated ubiquitination of NUMB protein: Identification of a second physiological substrate of MDM2 that employs a dual-site docking mechanism. 2012 J. Biol. Chem. 287, 14052–68.
Protein-protein interaction studies on protein arrays: effect of detection strategies on signal-to-background ratios. 2009 Anal. Biochem. 392, 45-53.
ATP modulation of Ca²⁺ release by type-2 and type-3 inositol (1, 4, 5)-triphosphate receptors. Differing ATP sensitivities and molecular determinants of action. 2008 J. Biol. Chem. 283, 21579–87.
Exploration of human ORFeome: High-throughput preparation of ORF clones and efficient characterization of their protein products. 2008 DNA Research 15, 137-149.
More Citations»

FAQs
Cloning Vectors FAQ

Software Tools
Flexi Vector Primer Design Tool

Use Restrictions

C8461, C8471, C8511, C8521, C8531, C8541, C8441, C8451 For Research Use Only. Not for Use in Diagnostic Procedures.

Patents - Disclaimers

All products on this page: Patent Pending.

All products on this page: For research use only. Persons wishing to use this product or its derivatives in other fields of use, including without limitation, commercial sale, diagnostics or therapeutics, should contact Promega Corporation for licensing information.

All products on this page: European Pat. No. 1685247 and other patents pending.

Cat.# C8461, C8471: This product or portions thereof is manufactured under license from Amrad Corporation Limited. For non-commercial research use only. All other uses require a license from Amrad Corporation Limited, Unit 6, 663 Victoria Street, Abbotsford, Victoria 3067, Australia, under U.S. Pat. No. 5,654,176, Australian Pat. No. 607511, Canadian Pat. No. 1338903 and other issued patents.

All products on this page: Usage restrictions apply to Bacterial Strains JM109(DE3), BL21(DE3)pLysS and KRX and to any derivatives thereof.

Usage Restrictions for the T7 Expression System
The T7 expression system is based on technology developed at Brookhaven National Laboratory under contract with the U.S. Department of Energy and is the subject of patents assigned to Brookhaven Science Associates, LLC (BSA). This technology, including bacteria, phage and plasmids that carry the gene for T7 RNA polymerase, is to be used for academic or nonprofit laboratory or licensed commercial research purposes only. By accepting or using the T7 expression technology you agree to be bound by the following conditions set forth by BSA. The initial purchaser may refuse to accept the conditions of this notice by returning this product and the enclosed materials to Promega unused.

Academic and NonProfit Laboratories
No materials that contain the cloned gene for T7 RNA polymerase may be distributed further to third parties outside of your laboratory unless the recipient receives a copy of this assurance notice and agrees to be bound by its terms. This limitation applies to Bacterial Strains JM109(DE3), BL21(DE3)pLysS and KRX and to any derivatives thereof.

Commercial Laboratories
A license is required for any commercial use of the T7 expression system, including use of the T7 system for research purposes or for production purposes by any commercial entity. Information about commercial licenses may be obtained from the Licensing Office, Brookhaven National Laboratory, Upton, NY 11973, Telephone: 631-344-7134, FAX: 631-344-3729.