Part:BBa_M0050:Design

AANDENYALAA. (Very fast) SsrA degradation tag.

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]

Design Notes

C-terminal tag. Degradation rate is very fast. Deviations from this sequence in key amino acids will lower degradation rates (see Parts BBa_M0051, BBa_M0052). Please realize that many variations of the WT SsrA sequence have been studied which may yield more desirable degradation rates; I suggest you consult the literature before deciding on the tag to use for your protein(see main page for refs). Sequence derived from reverse translation of AANDENYALAA sequence using codon usage optimized for E. coli. Three C-terminal aa's (LAA in this case) are necessary and sufficient for ClpX binding and degradation. Upstream aa sequence serves as a binding site for SspB, which guides rapid binding to ClpX. See sources on the main part page for further information about the mechanism of the system.

NOTE: this sequence has only the amino acid sequence for the tag and bears NO STOP CODONS, so be sure to inlcude them if you use this sequence.

Source

C-terminal degradation tags are commonly found in high turnover proteins in Escherichia coli.

References

See the following references for further information on degradation rates and mechanisms of this tag: (1)McGinness, Baker, Sauer. 2006. Mol. Cell. 22:701. (2)Flynn et al 2003. Mol. Cell. 11: 671. Flynn et al. 2001. PNAS 98(19): 10584. Anderson et al 1998. App. Env. Microbiol. 64(6):2240.