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Flexi® Cloning System

Flexi® Cloning System

Directional Cloning of Protein-Coding Sequences

  • Choose from a variety of initial applications (e.g., bacterial protein, mammalian or cell-free protein expression) and then transfer the insert as required
  • Efficient insert transfer means direct use of recombinant clones with minimal time for screening background colonies
  • Adaptable to high-throughput formats for large screening projects

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Description

Choose a kit

Size: 5 entry and 20 transfer reactions
Catalog number selected: C8640

Size: 50 transfer reactions
Catalog number selected: C9320

Size: 100 transfer reactions
Catalog number selected: C8820

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.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.

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

Complete Protocol

Flexi® Vector Systems Technical Manual


Specifications

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

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

BB

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

BB

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.