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FlexClone Collection
The study of gene function is critically dependent on the availability of gene expression vectors suitable for protein production. OriGene has long provided every member of its TrueClone collection of full-length cDNA clones in the pCMV vector suitable for transient expression in mammalian cells. We are now offering our TrueClones in other expression vectors based upon the robust shuttle vector platform from Promega called Flexi® Vector. These FlexClones can be located through the search bar above. The initial OriGene FlexClones are offered in Flexi® Vector GST (engineered for protein purification) and neoCMV constructs (optimized for stable cell line generation). The Flexi® Vector is a open-license restriction based cloning system which offer efficient, high-fidelity transfer between multiple expression vector cloning backbones without the need for additional sequence verification. The Flexi (a,b,c)Systems use two rare cutting restriction enzymes Sgf I (d) and Pme I (c) in a simple yet powerful directional cloning method for protein coding sequences. These systems provide a rapid efficient and high fidelity way to transfer between a variety of Flexi vectors. Flexi vectors carry the lethal barnase gene which is replaced by the DNA fragment of interest and acts as positive selection for ligations of the insert. To transfer your protein coding region from one Flexi Vector to another choose the appropriate acceptor vector with the desired expression and tag options the donor and acceptor vectors are digested with the Flexi enzyme blend simultaneously prior to ligation of the insert transformation and selection of cells.
Figure 1: Transferring protein-coding regions in the Flexi Vector Systems. The Flexi Vector Systems use a flexible directional cloning method for expressing protein coding regions with or without peptide fusion tags The features necessary for expression and any fusion tag are carried on the vector backbone and the protein-coding region can be shuttled between vectors using two rare cutting restriction enzymes Sgf I and Pme I The Flexi Vectors contain a lethal gene, barnase for positive selection of the protein coding sequences and antibiotic resistance marker for selection of colonies containing the Flexi vector a) Patent Pending. For information on Flexi® Vectors and related applications for the functional characteriztion of protein-coding regions, please visit the Promega site.
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