Protein_Domain

Part:BBa_K2356001:Design

Designed by: Ralf Philipsen   Group: iGEM17_TU-Eindhoven   (2017-10-26)


14-3-3 tetramer with GFP


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1306
    Illegal EcoRI site found at 1461
    Illegal PstI site found at 521
    Illegal PstI site found at 3967
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1306
    Illegal EcoRI site found at 1461
    Illegal NheI site found at 1642
    Illegal PstI site found at 521
    Illegal PstI site found at 3967
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1306
    Illegal EcoRI site found at 1461
    Illegal BamHI site found at 3229
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1306
    Illegal EcoRI site found at 1461
    Illegal PstI site found at 521
    Illegal PstI site found at 3967
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1306
    Illegal EcoRI site found at 1461
    Illegal PstI site found at 521
    Illegal PstI site found at 3967
    Illegal AgeI site found at 3958
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1009
    Illegal SapI.rc site found at 2602


Design Notes

Flexible linkers were added between the the domains, so that the protein domains have enough moving space.
The DNA sequence of the third and fourth monomers are scrambled, so that the DNA sequence is different, but the protein sequence remains similar. This allows for specific mutations on one of the monomers, without altering the other one.


Source

The 14-3-3 protein sequence originates from tobacco plants and the DNA sequence has been optimized for expression in E. coli. GFP originates from the jellyfish "Aequorea victoria".

References