Part:BBa_K4316029

NucA CDS + Sv40 NLS B5

The NucA nuclease and Sv40 part was designed to be used with the Chlamydomonas Reinhardtii MoClo kit [1] and is in position B5. Its 3' tag is ggTTCG as to maintain a correct reading frame.

The Sv40 NLS addresses proteins inside the cell nucleus. [3]

The nuclease A (NucA), a non-specific endonuclease, provokes cell death when localizing to the nucleus.[2]

The sequence of this part was retrieved from Davis et al. and Crozet et al. [4, 1]

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] Ghosh M, Meiss G, Pingoud A, London RE, Pedersen LC. Structural Insights into the Mechanism of Nuclease A, a ββα Metal Nuclease from Anabaena*. Journal of Biological Chemistry. 2005;280: 27990–27997. doi:10.1074/jbc.M501798200

[3] Lu J, Wu T, Zhang B, Liu S, Song W, Qiao J, et al. Types of nuclear localization signals and mechanisms of protein import into the nucleus. Cell Communication and Signaling. 2021;19: 60. doi:10.1186/s12964-021-00741-y

[4] Davis A, Moore IB, Parker DS, Taniuchi H. Nuclease B. A possible precursor of nuclease A, an extracellular nuclease of Staphylococcus aureus. J Biol Chem. 1977 Sep 25;252(18):6544-53. PMID: 893427.

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]