Part:BBa_K2333004
mf-Lon Protein Degradation Tag D (medium)
This protein degradation tag (pdt) is degraded by the Lon protease from Mycoplasma florum (mf-Lon) (BBa_K2333011). The Lon protease efficiently recognize and degrade C-terminal ssrA-tagged proteins, functioning in a similar way as ClpXP protease in most gram-negative bacteria. This Lon protease system is orthogonal to E. coli’s endogenous protein degradation machinery.
This tag is part of a series of 6 tags (labeled A thru F). pdt A is used as the parental tag to create tags B-F by changing the pdt residues 13–15 for mutagenesis because this region is essential for Lon-mediated degradation. Of this series of tags, this tag is relatively weak, and has a slow degradation rate. BBa_K2333001-Bba_K2333006 comprise this tag series. See Collins et al. 2014 "Tunable Protein Degradation in Bacteria" for the original design via mutagenesis, and for their characterization of degradation rates.
These parts are available in the William and Mary UNS format of: Tag-B0015(Double Terminator)-DoubleStop Codon which enables easy cloning using Gibson, Golden Gate or 3A assembly. See part series K2333401-K2333406 or BBa_K2333404 for this degradation tag.
Note: pdt D varies from pdt A in that residues 13-15 change from “PTF” to “AQP”.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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