Coding

Part:BBa_K4194008

Designed by: Jiong Huang   Group: iGEM22_BUCT-China   (2022-09-24)


RFP from Discosoma striata (coral) with LVA-ssrA degradation tag

Mutant RFP from Discosoma striata (coral) with added LVA-ssrA degradation tag. The tag increases RFP turn-over rate, thus providing better temporal resolution of red fluorescence. In the same time, maximal fluorescence amplitudes will be lower as newly formed protein is degraded as soon as it is formed.


Usage and Biology

ssrA-LVA tag

In E. coli [wild-type (sspB+ clpX+)], the adaptor sspB binds ssrAtagged substrates to ClpXP proteases, resulting in a modest increase in their degradation rate.
LVA tag is commonly found attached to repressors in various gene networks (e.g. oscillators). The tag encodes peptide sequence AANDENYALVA and is recognized by ClpA and ClpX unfoldases and ClpX mediator SspB. ClpA and ClpX then form a proteosome-like complex with ClpP protease and the protein is degraded.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 673
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 555
  • 1000
    COMPATIBLE WITH RFC[1000]


Functional Parameters

colorred
emission607
excitation584
tagssrA-LVA tag
The bacteria solution is red because of the expression of mRFP1.
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Categories
Parameters
colorred
emission607
excitation584
tagssrA-LVA tag