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T7 Vectors for Highest Expression Levels in Bacteria
- FLAG® 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
- lac operator to reduce "leaky" expression
- Enterokinase removal of N-terminal FLAG tags
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View a Vector Selection Guide.
| Product |
MCS Region |
P1118,
pT7-FLAG™-1
N-terminal Met-FLAG |
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P1243,
pT7-FLAG™-2
C-terminal FLAG |
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E5780,
pT7-MAT™-1
N-terminal MAT |
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E5655,
pT7-MAT™-2
C-terminal MAT |
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E5280,
pT7-FLAG-MAT™-1
N-terminal Met-FLAG, C-terminal MAT (dual tag) |
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E4905,
pT7-MAT™-FLAG-2
N-terminal MAT,C-terminal FLAG (dual tag) |
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E5155,
pT7-MAT-FLAG™-1
N-terminal MAT-FLAG (dual tag) |
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E5030,
pT7-FLAG-MAT™-2
C-terminal FLAG-MAT (dual tag) |
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- Kanamycin gene and ampicillin gene for greater selection flexibility in E. coli
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View a Vector Selection Guide.
| Product |
MCS Region |
P9618,
pT7-FLAG™-3
N-terminal Met-FLAG |
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P9743,
pT7-FLAG™-4
C-terminal FLAG |
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T7 Vector Features
Bacterial T7 promoter-based vectors allow for expression, detection and purification of recombinant FLAG and MAT (Metal Affinity Tag) fusions in E. coli. Several vectors containing the T7 promoter offer dual tag options for FLAG and MAT-tagged fusion proteins. These vectors confer ampicillin resistance for easy selection of positive transformants. Additionally, the vectors contain the T1T2 transcriptional terminator, the pMB1 (derivative of pBR322) origin of replication, the f1 origin and the lacI gene for repression of the T7 promoter.
pT7-FLAG™ and pT7-MAT™ vectors offer the very strong T7/lac promoter. These expression vectors produce even higher yields of recombinant protein than the tac promoter system. However, the T7 promoter is known for background ("leaky") expression, which can be a drawback when recombinant proteins are toxic to the host cell. Therefore, Sigma's vectors contain the lac operator (lacO) sequences immediately downstream from the promoter to reduce leaky expression. Unlike the tac promoter system, pT7 vectors must be expressed in hosts containing a source of the T7 polymerase such as (DE3) lysogenic strains.
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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