Part:BBa_K365006

LAA tag

This is the C-terminal end of the SsrA tag from E. coli. The N-terminal part of the SsrA tag is recognized by the adaptator protein SspB, wherease the C-terminal part (of which this biobrick is composed) is recognized by ClpX. This Biobrick is supposed to be at the C-terminal end of the protein to be tagged, thus there is NO fusion suffixe in this biobrick.

Background:

LAA tag is a C-terminal region of the natural ssrA-recognition sequence of E. Coli that interacts with the ClpXP protease. A protein fused with this tag will be preferentially degraded by the ClpX protease without need of an adaptor protein Baker and Sauer (2006). This tag serves as positive control for the functionality of the composed ClpXP and the PhyB-ClpXP fusion protein.


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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]

Conception:

We chose to use the same tag, which was already successfully used by used by Baker and Sauer (2006). It is a eleven amino acid tag (AANDENYALAA). At the end of the coding sequence a double stop codon was added (ACTAGT). The codon usage of E. coli (http://www.geneinfinity.org/sp_codonusage.html) has been used to decide the DNA sequence and NgoMIV and Pst1 restriction sites have been added to the sequence.