Tag

Part:BBa_K2333001

Designed by: Ethan M Jones   Group: iGEM17_William_and_Mary   (2017-06-14)

mf-Lon Protein Degradation Tag A (strong)


This protein degradation tag (pdt) is degraded by the AAA+ Lon protease from Mesoplasma florum (mf-Lon) (BBa_K2333011). While mf-Lon is evolutionarily related, and mechanistically similar to the E. coli Lon protease, mf-Lon is unable to recognize E. coli tags (commonly known as ssRA tags), and vice versa.

This tag is part of a series of 6 tags (labeled A thru F). Of this series of tags, this tag is the strongest (highest degradation rate). BBa_K2333001-Bba_K2333006 comprise this tag series. 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 K2333401 for this degradation tag.

See Collins et al. 2014 "Tunable Protein Degradation in Bacteria" for the original design of these tags via mutagenesis, and for their characterization of degradation rates.

Note: Each pdt in the series BBa_K2333001-Bba_K2333006 (pdt A-pdt F) is identical except for mutations on residues 13-15, which is the region of mf-Lon recognition.

Describe various degradation rates with parts here.


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]


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Parameters
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