Composite

Part:BBa_K3365054

Designed by: Zou Aixuan   Group: iGEM20_SJTU-BioX-Shanghai   (2020-10-22)


enhanced GFP with double terminator

This part is used as a reporter for detecting whether dCas9-ωis off-target. We inserted this part downstream of transcription activating unit of each lure sequence. If dCas9-ω is binding to the lure sequence, the expression of this part will be activated and the cell then emit green fluorescence, allowing us to know that dCas9-ωis off-target. As we plan to put the potential off-target site one by one, we added a terminator to eGFP so that once binding, the dCas9-ωcan only activating the closest reporter’s expression downstream.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 721
    Illegal XhoI site found at 730
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None