Composite

Part:BBa_K2477014

Designed by: Jacob Premo   Group: iGEM17_UMaryland   (2017-11-01)


Cas9/sgRNA GFP Mutant Selection System

This part contains the machinery to express Cas9 and an sgRNA targeting the chromophore region of GFP (E0040). The expression of both components of this system are under control of tetracycline promoters. The part also encodes for constitutive expression of the TetR repressor. This thereby allows for tight regulation of Cas9 and sgRNA expression for controlled mutant selection. This part is meant for use alongside a GFP variant containing an artificially induced PAM site (BBa_K2477003).

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2049
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1140
    Illegal SapI.rc site found at 2130


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Parameters
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