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Reporter

Part:BBa_K1399017

Designed by: Anna Stikane   Group: iGEM14_Edinburgh   (2014-09-19)

Plac-GFP(DAS+2)

Lactose/IPTG inducible promoter with GFP reporter tagged with DAS+2-ssrA degradation tag followed by terminator. The tagged GFP is actively degraded within cell (Figure 1), thus provides better temporal resolution of green 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.

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 870


[edit]
Categories
//cds/reporter/gfp
//classic/generator/prct
//classic/reporter/pret
//function/reporter/fluorescence
Parameters
emissionGFP
excitation
tagSsrA-DAS+2 degradation tag