Part:BBa_K2549021

aCD19-mN1c-tTAA

This part is one of our SynNotch receptors, similar to the original published version. It is our favorite. αCD19 (Part:BBa_K2549005) is used as the extracellular sensor module to receive the signal input from surface-expressed CD19. mN1c (Part:BBa_K2549006) is served as the transmembrane core domain of SynNotch, which is evident to have a low basal expression and a high activation efficiency. tTAA (Part:BBa_K2446057) is an improved tetracycline-controlled transactivator, which is cleaved after SynNotch activation and drives the expression of the amplifier. Besides, a CD8α signal peptide (Part:BBa_K2549044) and a Myc-tag (Part:BBa_K823036) are added to the N terminal of αCD19 (Part:BBa_K2549005) for membrane targeting and easy determination of surface expression.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 2493
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 534
  Illegal SapI.rc site found at 1408
  

Biology

It works as expected

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SynNotch receptors function well in Morsut L et al 2016

Morsut L et al stated:SynNotch receptors provide extraordinary flexibility in engineering cells with customized sensing/response behaviors to user-specified extracellular cues.

Morsut L et al have shown that modularity of the synNotch platform. They stated: the input and output domains from Notch can be swapped with diverse domains. On the extracellular side, diverse recognition domains can be used (antibody based, or peptide tags are shown) and on the intracellular side, diverse effector can be used (transcriptional activators with different DNA-binding domains are shown, as well as a transcriptional repressor).

Please refer to the original article for more details.