Plasmid

Part:BBa_K1160002

Designed by: Yang Rongwu   Group: iGEM13_NJU_NJUT_China   (2013-09-14)


cas9 cleavage detector

This plasmid is used to detect the site-specific cleavage activity conducted by Cas9 (BBa_K1160000) together with chimeric RNA (BBa_K1160001). This part contains a eGFP coding sequence that followed by the targeted sequence. (The targeted sequence is from the spacer sequence from E.coli str.K12, substr.W3110) Also, the targeted sequence is replaceable with the right restriction enzyme. If the cas9 protein authentically conduct the site-specific cleavage activity, this plasmid will be broken with a few nucleotides cut off from it. Thus the eGFP can not be expressed in the organism, which can be a detector of the cleavage.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 143
    Illegal EcoRI site found at 476
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 143
    Illegal EcoRI site found at 476
    Illegal NotI site found at 5080
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 143
    Illegal EcoRI site found at 476
    Illegal BglII site found at 5088
    Illegal BamHI site found at 1771
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 143
    Illegal EcoRI site found at 476
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 143
    Illegal EcoRI site found at 476
    Illegal NgoMIV site found at 4654
    Illegal NgoMIV site found at 4715
    Illegal AgeI site found at 1490
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2944
    Illegal BsaI.rc site found at 1754
    Illegal BsaI.rc site found at 4077


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