Tag

Part:BBa_M0051:Design

Designed by: Felix Moser   Group: Endy Lab   (2007-12-05)
Revision as of 18:21, 5 December 2007 by Fmoser76 (Talk | contribs) (→‎References)

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AANDENYNYADAS. (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 fast. Variations of this sequence yield different degradation rates(see Parts BBa_M0050, 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 AANDENYNYADAS sequence using codon usage optimized for E. coli. Three C-terminal aa's (DAS 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 inlclude them if you use this sequence.

Source

This variation of the SsrA tag was studied by McGinness, Baker, Sauer. 2006. Mol. Cell. 22:701.

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.