Composite

Part:BBa_K5317012:Design

Designed by: Anna Seidler   Group: iGEM24_Hannover   (2024-09-16)


CMV-MTF1-mRuby2


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1785
    Illegal PstI site found at 1030
    Illegal PstI site found at 1362
    Illegal PstI site found at 1819
    Illegal PstI site found at 1886
    Illegal PstI site found at 1924
    Illegal PstI site found at 2341
    Illegal PstI site found at 2671
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1785
    Illegal PstI site found at 1030
    Illegal PstI site found at 1362
    Illegal PstI site found at 1819
    Illegal PstI site found at 1886
    Illegal PstI site found at 1924
    Illegal PstI site found at 2341
    Illegal PstI site found at 2671
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1785
    Illegal BamHI site found at 1220
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1785
    Illegal PstI site found at 1030
    Illegal PstI site found at 1362
    Illegal PstI site found at 1819
    Illegal PstI site found at 1886
    Illegal PstI site found at 1924
    Illegal PstI site found at 2341
    Illegal PstI site found at 2671
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1785
    Illegal PstI site found at 1030
    Illegal PstI site found at 1362
    Illegal PstI site found at 1819
    Illegal PstI site found at 1886
    Illegal PstI site found at 1924
    Illegal PstI site found at 2341
    Illegal PstI site found at 2671
    Illegal NgoMIV site found at 2599
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 2710


Design Notes

The CMV promoter and EGFP reporter sequence were already part of the pEGFP-C2 backbone. The MTF-1 originated from murine NIH3T3 cDNA so it would exclude introns.


Source

The proteins contained in the CMV-MTF1-mRuby2 cassette have to be in frame. To fuse the mRuby2 reporter to the MTF-1 transcription factor it was important to cut the stop codon at the end of the MTF-1.

References