Science Catalog 2012 Life Cell Signaling Flexi® Cloning System Product Flexi® System, Entry/ Transfer Flexi® System, Transfer Carboxy Flexi® System, Transfer Available Separately 10X Flexi® Enzyme Blend (SgfI & PmeI) Carboxy Flexi® Enzyme Blend (SgfI & EcoICRI) HaloTag® Cloning Starter System For Research Use Only. Not for Use in Diagnostic Procedures. SgfI Description: 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. 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. Features: • Versatility: You can choose between a variety of initial applications (e.g., bacterial protein, mammalian, or cell-free protein expression) and then transfer to others as required. • Time Savings: Efficient transfer allows for direct use of recombinant clones, minimizing time wasted screening background colonies. • Enhanced Productivity: Adaptable to high-throughput formats for large screening projects. • Easy Access: No licensing fees or complicated transfer restrictions. Storage Conditions: Cat.# C8640 is comprised of Cat.# C8641 and A9280. Store Cat.# C8641 at –20°C; store Cat.# A9280 at room temperature. Store Cat.# C8820 and C9320 at –20°C. Store enzyme blends at –20°C. Protocol Flexi® Vector Systems Technical Manual Part# TM254 Ligation Size Cat.# Price (Fr) 5 entry and 20 transfer reactions C8640 416.00 100 transfer reactions C8820 1351.00 50 transfer reactions C9320 734.00 DNA Purification 25 µl R1851 161.00 100 µl R1852 564.00 50 µl R1901 161.00 1 each G6050 648.00 Digestion with Flexi® Enzyme Blend DNA Purification PmeI lethal gene SgfI lethal gene Acceptor Flexi® Vector PmeI SgfI PCR PmeI SgfI protein-coding region PmeI Digestion with Flexi® Enzyme Blend Transformation and Selection protein-coding region SgfI PmeI 98 For complete and up-to-date product information visit: www.promega.com/catalog 4599MA Pagina 101
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