{"id":1814,"date":"2021-06-05T16:47:52","date_gmt":"2021-06-05T16:47:52","guid":{"rendered":"https:\/\/www.embl.org\/groups\/protein-expression-purification\/?page_id=1814"},"modified":"2022-01-25T16:58:10","modified_gmt":"2022-01-25T16:58:10","slug":"e-coli-expression-vectors","status":"publish","type":"page","link":"https:\/\/www.embl.org\/groups\/protein-expression-purification\/services\/strategy-and-construct-design\/e-coli-expression-vectors\/","title":{"rendered":"E. coli expression vectors"},"content":{"rendered":"\n
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Overview of our bacterial expression vectors<\/a><\/p>\n\n\n\n

Please note that only the vectors indicated in yellow can be shared with external users via the EMBL MTA.<\/p>\n\n\n\n

The pETM<\/strong> 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>.<\/p>\n\n\n\n

A similar collection of E. coli<\/em> 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<\/a><\/strong> vectors and are specifically adapted for highly efficient ligation-independent cloning<\/strong>.<\/p>\n\n\n\n

The detailed cloning protocols and descriptions of these vectors can be found in the following reference:<\/p>\n\n\nScholz 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<\/em>. 13<\/strong>:12<\/blockquote>\n\n\n\n

Internal EMBL users can obtain these vectors at the Protein Expression and Purification Core Facility. External users can request them via Addgene<\/a>.<\/p>\n\n\n\n

Bacterial pCOOFY expression vectors<\/h3>\n\n\n\n
Name<\/th>N-terminal tag<\/th>C-terminal tag<\/th>Antibiotic resistance<\/th><\/tr><\/thead>
pCoofy1<\/td>His6<\/sub>-3C<\/td>X<\/td>Kanamycin<\/td><\/tr>
pCoofy2<\/td>Trx- His6<\/sub>-3C<\/td>X<\/td>Kanamycin<\/td><\/tr>
pCoofy4<\/td>His6<\/sub>– MBP-3C<\/td>X<\/td>Kanamycin<\/td><\/tr>
pCoofy5<\/td>His6<\/sub>-thrombin-hSumo1<\/td>X<\/td>Kanamycin<\/td><\/tr>
pCoofy6<\/td>His6<\/sub>-thrombin-hSumo3<\/td>X<\/td>Kanamycin<\/td><\/tr>
pCoofy7<\/td>S-tag-3C<\/td>X<\/td>Kanamycin<\/td><\/tr>
pCoofy8<\/td>HaloTag-3C<\/td>X<\/td>Kanamycin<\/td><\/tr>
pCoofy9<\/td>StrepII-3C<\/td>Thrombin-His6<\/sub><\/td>Ampicillin<\/td><\/tr>
pCoofy12<\/td>TwinStrepII-3C<\/td>X<\/td>Kanamycin<\/td><\/tr>
pCoofy16<\/td>His10<\/sub>-NusA-3C<\/td>X<\/td>Kanamycin<\/td><\/tr>
pCoofy18<\/td>His10<\/sub>– 3C<\/td>X<\/td>Kanamycin<\/td><\/tr>
pCoofy21<\/td>His10<\/sub>-TwinStrepII-3C<\/td>X<\/td>Kanamycin<\/td><\/tr>
pCoofy24<\/td>CBP-3C<\/td>His10<\/sub><\/td>Kanamycin<\/td><\/tr>
pCoofy26<\/td>NusA-3C<\/td>His10<\/sub><\/td>Kanamycin<\/td><\/tr>
pCoofy31<\/td>TwinStrepII-3C<\/td>His6<\/sub><\/td>Kanamycin<\/td><\/tr>
pCoofy33<\/td>His10<\/sub>– 3C<\/td>TwinStrepII<\/td>Kanamycin<\/td><\/tr>
pCoofy34<\/td>S-tag-TwinStrepII-3C<\/td>X<\/td>Kanamycin<\/td><\/tr>
pCoofy35<\/td>MBP-3C<\/td>X<\/td>Kanamycin<\/td><\/tr>
pCoofy36<\/td>S-tag-TwinStrepII-3C<\/td>His10<\/sub><\/td>Kanamycin<\/td><\/tr>
pCoofy37<\/td>MBP-3C<\/td>His6<\/sub><\/td>Kanamycin<\/td><\/tr>
pCoofy49<\/td>His6<\/sub>– MBP-3C<\/td>*His10<\/sub>
*TwinStrepII
*S-tag
*CBP<\/td>		Kanamycin<\/td><\/tr>
pCoofy52<\/td>His6<\/sub>– Avi-Avi-Avi-3C<\/td>X<\/td>Kanamycin<\/td><\/tr>
pCoofy61<\/td>His6<\/sub>– eGFP-3C<\/td>X<\/td>Kanamycin<\/td><\/tr><\/tbody><\/table>
* Multiple options for the C-terminal tag depending on the cloning strategy<\/figcaption><\/figure>\n\n<\/div>\n<\/div>\n\n\n
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