Part:BBa_K133095

CMV-SS-HA-multiHP-CD4ecto-(TMTIR)TLR3-GFP-Histop

A synthetic Helicobacter pylori multiepitope antigen fused with the extracellular domain of human CD4 and the transmembrane (TM) and 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 protein can be visualized by fluorescent microscopy, because it contains GFP on the C-terminal side, before the His-tag. It is also possible to lable 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 3013
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

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