Difference between revisions of "Part:BBa K1399013"

 
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<partinfo>BBa_K1399013 short</partinfo>
 
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Lactose/IPTG inducible promoter with RFP reporter tagged with DAS-ssrA degradation tag followed by terminator.
 
Lactose/IPTG inducible promoter with RFP reporter tagged with DAS-ssrA degradation tag followed by terminator.
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The tagged RFP is degraded within cell, thus provides better temporal resolution of red fluorescence and promoter activity. Presence of SspB is crucial for effective protein degradation.
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The tagged RFP is degraded within cell (Figure 1), thus provides better temporal resolution of red fluorescence and promoter activity. Presence of SspB is crucial for effective protein degradation.
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[[File:EDiGEM14-SsrA degron mediated degradation.png|400px|thumb|center|'''Figure 1 SsrA degron mediated protein degradation.''' ('''A''') Any version of SsrA tags can be attached to any protein of interest (e.g. RFP or GFP). ('''B''') The tag is recognized by ClpX unfoldase forming a complex with ClpP protease and the tagged protein gets degraded.]]
  
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===Usage and Biology===
  
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[http://2017.igem.org/Team:Kingsborough_NY Team Kingsborough NY 2017] contributed to the characterization of this part by showing decreased fluorescence when expressed either in a higher salt media - such as LB with 3% sodium chloride - or E. coli that lacks tmRNA, the principal component of the cell's ribosome rescue system. Cells without tmRNA will have less native ssrA substrates, and this may lead to increased degradation of ssrA tagged RFP. For more information and data, see the experience page or [http://2017.igem.org/Team:Kingsborough_NY/RFP visit our Wiki]<br>
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===Usage and Biology===
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(<small>--[[User:djcamenares|djcamenares]] 17:56, 27 October 2017 (UTC) </small>)
  
 
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Latest revision as of 19:55, 19 November 2017

Plac-RFP(DAS)

Lactose/IPTG inducible promoter with RFP reporter tagged with DAS-ssrA degradation tag followed by terminator. The tagged RFP is degraded within cell (Figure 1), thus provides better temporal resolution of red fluorescence and promoter activity. Presence of SspB is crucial for effective protein degradation.

Figure 1 SsrA degron mediated protein degradation. (A) Any version of SsrA tags can be attached to any protein of interest (e.g. RFP or GFP). (B) The tag is recognized by ClpX unfoldase forming a complex with ClpP protease and the tagged protein gets degraded.

Usage and Biology

[http://2017.igem.org/Team:Kingsborough_NY Team Kingsborough NY 2017] contributed to the characterization of this part by showing decreased fluorescence when expressed either in a higher salt media - such as LB with 3% sodium chloride - or E. coli that lacks tmRNA, the principal component of the cell's ribosome rescue system. Cells without tmRNA will have less native ssrA substrates, and this may lead to increased degradation of ssrA tagged RFP. For more information and data, see the experience page or [http://2017.igem.org/Team:Kingsborough_NY/RFP visit our Wiki]
(--djcamenares 17:56, 27 October 2017 (UTC) )

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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 781
    Illegal AgeI site found at 893
  • 1000
    COMPATIBLE WITH RFC[1000]