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CMV vectors for high-level expression
- Stable expression with neomycin selection
- FLAG® and 3xFLAG® vectors for highly sensitive detection and purification using ANTI-FLAG® antibodies, resins, and plates
- MAT™ vectors for high quality purification using HIS-Select™ resins and 96-well plates
- N- or C- terminal fusions
- Dual tag configurations
- Cytoplasmic expression or secretion
- Enterokinase removal of N-terminal FLAG tags
- Expression kits available
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View a Vector Selection Guide.
We also provide BICEP bicistronic expression vectors for stable expression.
| Product |
MCS Region |
E8770, pFLAG-CMV™-3
N-terminal FLAG (secreted) |
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E1775, pFLAG-CMV™-4
N-terminal Met-FLAG |
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E4276, p3xFLAG-CMV™-9
N-terminal 3xFLAG (secreted) |
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E4401, p3xFLAG-CMV™-10
N-terminal Met-3xFLAG |
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E4776, p3xFLAG-CMV™-13
C-terminal 3xFLAG (secreted) |
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E4901, p3xFLAG-CMV™-14
C-terminal 3xFLAG |
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E5776, pFLAG-myc-CMV™-21
N-terminal FLAG, C-terminal c-Myc (Dual tagged, secreted) |
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E5901, pFLAG-myc-CMV™-22
N-terminal Met-FLAG, C-terminal c-Myc (Dual tagged) |
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E6276, p3xFLAG-myc-CMV™-25
N-terminal 3xFLAG, C-terminal c-Myc (Dual tagged, secreted) |
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E6401, p3xFLAG-myc-CMV™-26
N-terminal Met-3xFLAG, C-terminal c-Myc (Dual tagged) |
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CMV vectors Features
CMV promoter-based vectors from Sigma-Aldrich provide flexibility in transient or stable expression, cytoplasmic expression or secretion, and N-terminal or C-terminal tagging in various combinations of FLAG, 3xFLAG, c-myc or MAT. These fusion proteins allow easy detection, purification and analysis of recombinant protein for a wide range of applications. Dual-tagged fusions provide flexibility in detection as well as a method to screen for full-length recombinant protein.
Our vectors for stable expression, as those for transient expression, contain the strong CMV promoter for high-level constitutive expression in mammalian cells. In addition, these constructs carry the aminoglycoside phosphotransferase II gene (neomycin resistance gene or neor) that confers resistance to aminoglycosides such as G 418 sulfate, allowing selection of stable transfectants. The strong human cytomegalovirus (CMV) promoter regulatory region drives constitutive protein expression levels as high as 1 mg/L in COS cells. For less potent cell lines, protein levels are typically ~0.1 mg/L. The presence of the SV40 replication origin will result in high levels of DNA replication in SV40 replication permissive COS cells. CMV vectors contain the pMB1 (derivative of pBR322) origin for replication in bacterial cells, the b-lactamase gene for ampicillin resistance selection in bacteria, hGH polyA, and the f1 origin. Vectors containing the preprotrypsin leader (PPT) sequence direct secretion of FLAG fusion proteins into the culture medium for purification using ANTI-FLAG antibodies, resins, and plates.
3xFLAG
The 3xFLAG epitope tag is 20-200 times more sensitive than the original FLAG tag. In cases of low-level expression, 3xFLAG is ideal. 3xFLAG is only 22 amino acids and is therefore unlikely to alter protein function or block other binding sites or epitopes. Like the original FLAG tag, 3xFLAG is very hydrophilic and can be cleaved with enterokinase.
MAT Tag
The MAT tag or Metal Affinity Tag (HNHRHKH) has been created for purification of recombinant MAT fusion proteins using HIS-Select Nickel and Cobalt Affinity Gels. HIS-Select products allow for highly selective purification of histidine-tagged fusion proteins such as MAT fusions. Many of our newest vectors make use of the MAT tag, often in combination with the well-known FLAG tag. MAT tag containing vectors are offered in formats for N-terminal or C-terminal tagging.
Enterokinase Removal of FLAG Tags
The recognition sequence for enterokinase, Asp-Asp-Asp-Asp-Lys, is found at the C-terminal end of the FLAG epitope tag. Removal of FLAG is possible in all fusion proteins containing an N-terminal FLAG sequence. Dual tag fusion proteins may also be cleaved with enterokinase for removal of one or more tags, depending on the position of FLAG in the protein sequence.
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