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Part:BBa_K3365001

Designed by: Zou Aixuan   Group: iGEM20_SJTU-BioX-Shanghai   (2020-10-22)
Revision as of 12:30, 27 October 2020 by Gyaxxxxxx (Talk | contribs)


Target binding site for dCas9 and subnit Omega

This part is used to detect whether dCas9 protein is on-target. The sequence is made up of PDCD1, which is the binding site of dCas9, and J23117 BBa_J23117 (visit:[1]) weak promoter spaced by 35 base pairs. The dCas9 in this part could bind to the PDCD1, under guidance of specific designed sgRNA, while subunit Omega could bind to J23117 and activating the expression of downstream fluorescent protein. According to reference, we knew that 35 base pairs is the best length of spacer for activating, so we randomly choose 35bp sequence from the plasmid pUC57 as spacer.

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
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None