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
Catalog Number:
Choose a kit
Size
Catalog Number: C8640
Catalog Number: C8820
Catalog Number: C9320
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.
Cloning a protein-coding region into the Flexi® Vectors.
Transfer of a protein-coding region between N-terminal or native Flexi® Vectors.
For more information about the HaloTag® Flexi® Vectors, please visit the HaloTag® Technology page.
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
Specifications
Catalog Number:
What's in the box?
| Item | 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
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
See Protocol for storage recommendations.
What's in the box?
| Item | 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 |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
See Protocol for storage recommendations.
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?
| Item | 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 |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
See Protocol for storage recommendations.
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.
Related Products
Similar Products
Frequently Used With
HaloLinkâ„¢ Resin
HaloTag® ligand bound to Sepharose beads that specifically and rapidly binds HaloTag® fusion proteins.
G1912, G1913, G1914, G1915
HaloTag® Standard Protein
A 61kDa purified HaloTag®-GST fusion protein supplied at a concentration of 3.0mg/ml. Use to estimate the expression level of your HaloTag® fusion protein.
G4491
BL21(DE3)pLysS Competent Cells
Ready-to-transform E. coli cells with IPTG-inducible protein expression from the T7 promoter.
L1195, L1191
TnT® Quick Coupled Transcription/Translation System
Coupled transcription/translation system that produces significant quantities of protein for applications including GST pull-downs and gel shift assays in 60–90 minutes.
L1170, L1171, L2080, L2081
MagneHisâ„¢ Protein Purification System
Purification of His- or HQ-tagged, proteins from a crude cell lysate on paramagnetic particles with either a manual (requires use of a magnetic stand) or automated procedure.
V8500, V8550
HaloTag® Fluorescent Ligands
For protein labeling and detection. Each includes a chloroalkane group for HaloTag® protein attachment.
G8281, G8282, G8591, G8592
Magne® HaloTag® Beads
Magnetic capture of HaloTag® fusion proteins for protein pull-downs and purification. Magnetics enable streamlined protein purification, eliminate multiple centrifugation steps.
G7281, G7282
Anti-HaloTag® pAb
Polyclonal rabbit antibody that specifically detects HaloTag® fusion proteins.
G9281
Anti-HaloTag® Monoclonal Antibody
Monoclonal antibody raised in mice against the HaloTag® protein for detecting HaloTag® fusion proteins in Western blotting.
G9211