Unlike site-specific recombination vector systems, Flexi systems do not require appending multiple amino acids to the amino or carboxy termini of the protein of interest. -Application: Directional Cloning Method for Protein-Coding Sequences -Systems Don't Require Appending Multiple Amino Acids to Protein Termini -Archival Entry Vector Not Required -Licensing Fees Not Required -Direct Entry for Most Applications Most applications allow direct entry into the vector most suited to experimental design. Researchers can choose between various initial applications and transfer to others as needed. Systems can adapt to high throughput formats for larger screening projects. Theyâre based on two rare-cutting restriction enzymes, SgfI and PmeI, to provide an efficient and high-fidelity way to transfer protein-coding regions between a variety of Flexi vectors without needing to resequence. All Flexi vectors carry the lethal barnase gene, which is replaced by the DNA fragment of interest and acts as a positive selection for successful ligation of the insert. C-terminal vectors allow expression of C-terminal-tagged proteins. The vectors can act as acceptors of protein-coding regions flanked by SgfI and PmeI, but they lack a PmeI site and have an EcoICRI blunt-ended site that canât be removed from the C-terminal Flexi vectors and transferred to other Flexi vectors. 0) also contains the carboxy Flexi enzyme blend (SgfI and EcoICRI).
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