Difference between revisions of "Part:BBa K2323003"
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===Usage and Biology===
 
===Usage and Biology===
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Tuning the degradation rate of parts is essential to construct dynamic circuits, such as oscillators or bistable switches. Our collection gathers five different tags with degradation kinetics ranging over one order of magnitude, so that they can be used for prototyping circuits’ phase space. The tags are taken from Andersen et al., New Unstable Variants of Green Fluorescent Protein for Studies of Transient Gene Expression in Bacteria, Applied and Environmental Microbiology, 64(6), June 1998 and Cameron & Collins, Tunable protein degradation in bacteria, Nature biotechnology, 32(12), December 2014.
  
 
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<partinfo>BBa_K2323003 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2323003 SequenceAndFeatures</partinfo>
  
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===Plasmid composition===
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[[File:BBa_K2323003.png| frame | 400px | center | The collection is cloned from [https://parts.igem.org/Part:BBa_I13522 BBa_I13522] with overhang PCR using 5'-phosphated primers that contain the tag as an overhang. Primers are indicated with the map ([https://benchling.com/s/y2WontDc read-only benchling map]). Cloning was confirmed with sequencing. ]]
  
 
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Revision as of 13:17, 30 October 2017


GFP-pdt2E

This GFP (E0040) is tagged with a protein degradation tag (pdt) called pdt2E. This causes the protein to degrade in E.coli with a first-order rate of 0.0046 per min, from the ClpP machinery of E.coli. It is under the control of a pTet promoter (aTc inducible) and the RBS is B0034. The terminator is B0015.

This biobrick is part of a collection of degradation tags, characterized on this page. See also BBa_K2323005, BBa_K2323006, BBa_K2323007, BBa_K2323008.

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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 724

Plasmid composition

The collection is cloned from BBa_I13522 with overhang PCR using 5'-phosphated primers that contain the tag as an overhang. Primers are indicated with the map (read-only benchling map). Cloning was confirmed with sequencing.