Composite

Part:BBa_K3365061

Designed by: Zou Aixuan   Group: iGEM20_SJTU-BioX-Shanghai   (2020-10-23)
Revision as of 11:07, 25 October 2020 by Gyaxxxxxx (Talk | contribs)


Target transcription activating unit upstream of RFP

This part is a on-target detecting platform for dCas9-ω. We put this part in a plasmid and transfer it into E.coli, which was knocked out rpoZ. For convenience, we called it △rpoZ-MG1655. If dCas9-ω is binding to target sequence, the submit Omega fused to dCas9 C-terminal can activate the expression of reporter by recruiting the RNAP holoenzyme and the cell would emit red fluorescence,which means dCas9-ωis on-target, else the weak promoter and the lack of rpoZ makes the reporter cannot express or just have a quite low expression level.

Target binding site for dCas9 and subnit Omega.png

Fig.1: Schematic diagram of this part

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 66
    Illegal NheI site found at 89
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 812
    Illegal XhoI site found at 803
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 667
    Illegal AgeI site found at 779
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None