Description
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The Flexi® Vector System is a 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 without the need to resequence. Once an ORF is cloned, conveniently transfer the inserts into a variety of vectors designed for multiple applications. No licensing fees or complicated transfer restrictions needed.
All Flexi® Vectors carry the lethal barnase gene, which is replaced by the DNA fragment of interest and acts as a positive selection for the successful ligation of the insert.
Unlike site-specific recombination vector systems, the Flexi® Vector Systems do not require appending multiple amino acids to the amino or carboxy termini of the protein of interest. In addition, the systems do not require an archival entry vector, and most applications allow direct entry into the vector suited to the experimental design.
C-terminal Flexi® Vectors allow expression of C-terminal-tagged proteins. While these vectors can act as acceptors of protein-coding regions flanked by SgfI and PmeI, they lack a PmeI site and contain a different blunt-ended site, EcoICRI. This joined sequence cannot be removed from the C-terminal Flexi® Vectors and transferred to other Flexi® Vectors.
Transfer of a protein-coding region between N-terminal or native Flexi® Vectors.
Transfer of a protein-coding region between N-terminal or native Flexi® Vectors.
Cloning a protein-coding region into the Flexi® Vectors.
Cloning a protein-coding region into the Flexi® Vectors.
Note: For access to 20,000 human and 15,000 mouse ORF clones with HaloTag® Technology in multiple mammalian expression vectors and custom cloning options, visit: www.genecopoeia.com and www.kasuza.or.jp.kop/dsearch-e
Protocols
What’s in the box?
Size: 5 entry and 20 transfer reactions
Item / Catlog no: C8640 | Part # | Size | Available Separately |
Flexi® Enzyme Blend (SgfI & PmeI) | R188A | 2 × 50μl | View Product |
T4 DNA Ligase (HC) | M179A | 1 × 500u | |
5X Flexi® Digest Buffer | R186A | 1 × 1ml | |
2X Flexi® Ligase Buffer | R187A | 1 × 500μl | |
Nuclease-Free Water | P119A | 2 × 1,250μl | View Product |
Wizard® SV Minicolumns | A129C | 1 × 10/pk | |
Collection Tubes | A130C | 1 × 10/pk | |
Vacuum Adapter | A133C | 1 × 5/pk | View Product |
Membrane Wash Solution | A929A | 1 × 3ml | |
Membrane Binding Solution | A930A | 1 × 4ml | View Product |
SDS
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.
Storage Conditions
See Protocol for storage recommendations.
What’s in the box?
Size: 50 transfer reactions
Item / Catlog no: C9320 | Part # | Size | Available Separately |
T4 DNA Ligase (HC) | M179A | 2 × 500u | |
Nuclease-Free Water | P119A | 1 × 1,250μl | View Product |
5X Flexi® Digest Buffer | R186A | 1 × 1ml | |
2X Flexi® Ligase Buffer | R187A | 1 × 500μl | |
Flexi® Enzyme Blend (SgfI & PmeI) | R188A | 1 × 50μl | View Product |
Carboxy Flexi® Enzyme Blend (SgfI & EcoICRI) | R190A | 1 × 50μl | View Product |
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.
Storage Conditions
See Protocol for storage recommendations.
Patents and Disclaimers
U.S. Pat. Nos. 8,293,503, 9,018,014 and 8,367,403, European Pat. No. 1685247 and other patents and patents pending.
Patent Pending.
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.
What’s in the box?
Size: 100 transfer reactions
Item / Catlog no: C8820 | Part # | Size | Available Separately |
T4 DNA Ligase (HC) | M179A | 4 × 500u | |
Nuclease-Free Water | P119A | 2 × 1,250μl | View Product |
5X Flexi® Digest Buffer | R186A | 1 × 1ml | |
2X Flexi® Ligase Buffer | R187A | 2 × 500μl | |
Flexi® Enzyme Blend (SgfI & PmeI) | R188A | 4 × 50μl | View Product |
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.
Storage Conditions
See Protocol for storage recommendations.
Patents and Disclaimers
U.S. Pat. Nos. 8,293,503, 9,018,014 and 8,367,403, European Pat. No. 1685247 and other patents and patents pending.
Patent Pending.
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.