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Protein Expression and Purification Core Facility

PEPCF expresses proteins in bacteria, insect and mammalian cells and uses a variety of chromatographic and biophysical techniques for protein purification and characterization.

E. coli expression vectors

Overview of our bacterial expression vectors

Please note that only the vectors indicated in yellow can be shared with external users via the EMBL MTA.

The pETM vectors developed at EMBL were designed in such a way that you can easily clone your gene of interest in a series of expression vectors with a large variety of fusion tags. This allows quick screening of a multitude of constructs to determine the most optimal fusion tags for your protein of interest. Except for the pETM11-Sumo3-eGFP vector, all pETM vectors have compatible multiple cloning sites to allow easy, fast and parallel cloning. Here you can find some more information on how to clone into the popular pETM11-Sumo3-eGFP vector.

A similar collection of E. coli vectors containing a variety of fusion tags was developed by Sabine Suppmann at the Max Planck Institute for Biochemistry in Martinsried, Munich. These vectors are called pCoofy vectors and are specifically adapted for highly efficient ligation-independent cloning.

The detailed cloning protocols and descriptions of these vectors can be found in the following reference:

Scholz J., Besir H., Strasser S. and Suppmann S. (2013) A new method to customize protein expression vectors for fast, efficient and background free parallel cloning. BMC Biotechnology. 13:12

Internal EMBL users can obtain these vectors at the Protein Expression and Purification Core Facility. External users can request them via Addgene.

Bacterial pCOOFY expression vectors

NameN-terminal tagC-terminal tagAntibiotic resistance
pCoofy1His6-3CXKanamycin
pCoofy2Trx- His6-3CXKanamycin
pCoofy4His6– MBP-3CXKanamycin
pCoofy5His6-thrombin-hSumo1XKanamycin
pCoofy6His6-thrombin-hSumo3XKanamycin
pCoofy7S-tag-3CXKanamycin
pCoofy8HaloTag-3CXKanamycin
pCoofy9StrepII-3CThrombin-His6Ampicillin
pCoofy12TwinStrepII-3CXKanamycin
pCoofy16His10-NusA-3CXKanamycin
pCoofy18His10– 3CXKanamycin
pCoofy21His10-TwinStrepII-3CXKanamycin
pCoofy24CBP-3CHis10Kanamycin
pCoofy26NusA-3CHis10Kanamycin
pCoofy31TwinStrepII-3CHis6Kanamycin
pCoofy33His10– 3CTwinStrepIIKanamycin
pCoofy34S-tag-TwinStrepII-3CXKanamycin
pCoofy35MBP-3CXKanamycin
pCoofy36S-tag-TwinStrepII-3CHis10Kanamycin
pCoofy37MBP-3CHis6Kanamycin
pCoofy49His6– MBP-3C*His10
*TwinStrepII
*S-tag
*CBP
Kanamycin
pCoofy52His6– Avi-Avi-Avi-3CXKanamycin
pCoofy61His6– eGFP-3CXKanamycin
* Multiple options for the C-terminal tag depending on the cloning strategy
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