Description_x000D_ General description_x000D_ General description_x000D_ pCMV-FLAG-MAT-Tag®-1 is a 4.7 kb mammalian expression vector used for the transient production of dual tagged fusion proteins in mammalian cells. This vector is a derivative of the pCMV5 transient expression vector for intracellular expression of a properly inserted open reading frame as a Met-N-terminal FLAG®, C-terminal MAT-Tag (Metal Affinity Tag) fusion protein._x000D_ The pCMV-FLAG-MAT-Tag-1 expression vector is a shuttle vector containing both bacterial and SV40 origins of replication for propagation in both E. coli and mammalian cells. Efficiency of replication in mammalian cells is optimal when using host cells that express the SV40 large T antigen. Removal of the FLAG tag is possible using enterokinase which cleaves following the Asp-Asp-Asp- Asp-Lys recognition site at the C-terminal end of FLAG._x000D_ The FLAG epitope of the fusion protein may be detected using Monoclonal ANTI-FLAG M2, Catalog Number F3165. The fusion protein may be purified using the ANTI-FLAG M2 Affinity Gel, Catalog Number A2220. The fusion protein may also be purified utilizing the metal affinity properties of the MAT-Tag by using HIS-Select® Nickel Affinity Gel, Catalog Number P6611._x000D_ Vector Maps and Sequences_x000D_ Application_x000D_ pCMV-FLAG-MAT-Tag-1 Expression Vector is suitable for transient cytoplasmic expression of dual-tagged fusion proteins under the CMV promoter._x000D_ Components_x000D_ pCMV-FLAG-MAT-Tag-1 Expression Vector, 20 μg (E1155) is supplied as 0.5 mg/ml in 10 mM Tris-HCl (pH 8.0) with 1 mM EDTA._x000D_ pCMV-FLAG-MAT-Tag-1-MAPK1 Control Vector, 20 μg (C0489) is supplied as 0.5 mg/ml in 10 mM Tris-HCl (pH 8.0) with 1 mM EDTA._x000D_ Principle_x000D_ The promoter-regulatory region of the human cytomegalovirus drives transcription of the fusion containing the FLAG® epitope (DYKDDDDK) and the transition metal binding MAT-Tag® (HNHRHKH)._x000D_ Legal Information_x000D_ FLAG is a registered trademark of Sigma-Aldrich Co. LLC_x000D_ HIS-Select is a registered trademark of Sigma-Aldrich Co. LLC_x000D_ MAT-Tag is a registered trademark of Sigma-Aldrich Co. LLC