Difference between revisions of "Part:BBa K2333001"
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| − | This protein degradation tag (pdt) is degraded by the AAA+ Lon protease from Mesoplasma florum (mf-Lon) < | + | This protein degradation tag (pdt) is degraded by the AAA+ Lon protease from Mesoplasma florum (mf-Lon) (<partinfo>Bba_K2333011</partinfo>). 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 | + | 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 <bbpart>K2333401</bbpart> 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. | 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 | + | 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. |
<b> Describe various degradation rates with parts here. </b> | <b> Describe various degradation rates with parts here. </b> | ||
| − | + | <!-- | |
| − | <!-- | + | |
===Usage and Biology=== | ===Usage and Biology=== | ||
| − | <!-- --> | + | While any mf-Lon generating part can be used alongside this tag to increase degradation rate/speed of a given protein of interest, the majority of William and Mary 2017's characterization was done using BBa_K2333434, which is a LacI regulated (IPTG inducible) mf-Lon. In cases where LacI cannot be used, the leakier Arabinose inducible mf-Lon <partinfo>BBa_K2333435</partinfo> can be used instead. (Note, it is recommended that these parts be used on a low copy backbone such as pSB3K3) <!-- --> |
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
<partinfo>BBa_K2333001 SequenceAndFeatures</partinfo> | <partinfo>BBa_K2333001 SequenceAndFeatures</partinfo> | ||
Latest revision as of 17:54, 29 October 2017
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]