Part:BBa_K5317012:Design

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.