Part:BBa_K4316023

TEV proteinase Nter fragment B5

The TEV (Tobacco Etch Virus) protease Nter fragment was designed to be used with the Chlamydomonas Reinhardtii MoClo kit [1] and is in position B5.

The TEV protease is a highly site-specific cysteine protease. Usually used to cleave tags from fusion proteins [2]. It was shown to be effective when 2 heterodimers, Nter and Cter split TEV proteases, were sterically joined. [3]

This sequence was retrieved from Wehr et al. [3].

Thymidine in position 216 was replaced with cytidine as to remove PstI site and make part compatible with RFC10 standard.

Sources:

[1] Crozet, Pierre & Navarro, Francisco & Willmund, Felix & Mehrshahi, Payam & Bakowski, Kamil & Lauersen, Kyle & Pérez-Pérez, María & Auroy, Pascaline & Gorchs Rovira, Aleix & Sauret-Gueto, Susana & Niemeyer, Justus & Spaniol, Benjamin & Theis, Jasmine & Trösch, Raphael & Westrich, Lisa & Vavitsas, Konstantinos & Baier, Thomas & Hübner, Wolfgang & de Carpentier, Félix & Lemaire, Stéphane. (2018). Birth of a Photosynthetic Chassis: A MoClo Toolkit Enabling Synthetic Biology in the Microalga Chlamydomonas reinhardtii. ACS Synthetic Biology. 7. 10.1021/acssynbio.8b00251.

[2] Raran-Kurussi S, Cherry S, Zhang D, Waugh DS. Removal of Affinity Tags with TEV Protease. In: Burgess-Brown NA, editor. Heterologous Gene Expression in E.coli. New York, NY: Springer New York; 2017. pp. 221–230. doi:10.1007/978-1-4939-6887-9_14

[3] Wehr MC, Laage R, Bolz U, Fischer TM, Grünewald S, Scheek S, Bach A, Nave KA, Rossner MJ. Monitoring regulated protein-protein interactions using split TEV. Nat Methods. 2006 Dec;3(12):985-93. doi: 10.1038/nmeth967. Epub 2006 Oct 29. PMID: 17072307.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]