Part:BBa_K511301

Notch-Gal4-VP16 Juxtacrine Signaling Receptor MammoBlock

This part encodes a major mammalian cell-signaling protein, the Notch-1 receptor, engineered to replace the original 12xCSL-binding transactvation domain with the Gal4-VP16 transactivator (Gal4-ESN). In the presence of the Delta ligand protein from another cell, this transactivation domain is cleaved and released to the Notch producing cell's nucleus. In contrast, if Delta is present on the Notch producing cell, the Delta protein will cis-inhibit the Notch receptor, desensitizing the receptor to activation from external Delta molecules. This receptor is singularly useful for cell patterning, lineage decision-making, and other forms of signaling-intensive processes.

This version of the Notch signaling protein is expected to induce much stronger levels of activation than those documented in recent work by Sprinzak, et al.

For more information, consult the MIT iGEM 2011 wiki here.

Sprinzak, D. et al. Cis interactions between Notch and Delta generate mutually exclusive signaling states. Nature 25 April 2010 http://www.nature.com/nature/journal/v465/n7294/full/nature08959.html

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 258
  Illegal PstI site found at 663
  Illegal PstI site found at 993
  Illegal PstI site found at 1224
  Illegal PstI site found at 1321
  Illegal PstI site found at 2511
  Illegal PstI site found at 4389
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 258
  Illegal PstI site found at 663
  Illegal PstI site found at 993
  Illegal PstI site found at 1224
  Illegal PstI site found at 1321
  Illegal PstI site found at 2511
  Illegal PstI site found at 4389
  Illegal NotI site found at 5224
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 2933
  Illegal XhoI site found at 5750
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 258
  Illegal PstI site found at 663
  Illegal PstI site found at 993
  Illegal PstI site found at 1224
  Illegal PstI site found at 1321
  Illegal PstI site found at 2511
  Illegal PstI site found at 4389
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 258
  Illegal PstI site found at 663
  Illegal PstI site found at 993
  Illegal PstI site found at 1224
  Illegal PstI site found at 1321
  Illegal PstI site found at 2511
  Illegal PstI site found at 4389
  Illegal NgoMIV site found at 524
  Illegal NgoMIV site found at 2636
  Illegal NgoMIV site found at 2692
  Illegal NgoMIV site found at 2735
  Illegal NgoMIV site found at 3520
  Illegal NgoMIV site found at 4537
  Illegal NgoMIV site found at 4714
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 5669
  Illegal BsaI.rc site found at 85
  Illegal BsaI.rc site found at 1801
  Illegal BsaI.rc site found at 2716
  Illegal BsaI.rc site found at 5173