Part:BBa_K133101

CMV-SS-HA-multiHP-CD4sh-(TIR)TLR3-Histop

A synthetic Helicobacter pylori multiepitope antigen fused with the extracellular and transmembrane domain of human CD4 and the cytosolic (TIR) domain of human Toll-like receptor 3. Construct is under control of constitutive promotor CMV and contains His-tag on C-terminus and HA-tag on N-terminus (after the CD4 signaling sequence).

Construct codes for a transmembrane protein expressed on the endosomal membrane. The construct can be visualized by labeling the protein with fluroscently labeled antibodies against the HA-tag inside the endosomes or against the His-tag in the cytosol.

The CD4 ecto domain enables constitutive homodimerization of the protein and thus activation of the TIR domain. This activation triggers transcription of genes under the IFNb promoter and stimulates the innate immune response. After the activation the antigen is processed in the endosome and short peptides are presented on MHC II molecules.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 1840
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 2939
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 2453
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 2287
  Illegal SapI.rc site found at 1375