Part:BBa_K133023

CMV-ss-CF213-multiHP-CF215-RGD-Histop (term)

We have used CMV promotor, because it is promotor for constitutive expression in mammalian cells.

SS is a CD4 signal peptide for trafficking to the cellular membrane.

This part is also composed of:

CF213 – the N-terminal portion of chimeric fusion protein; it contains 176 AA of the N-terminal portion of E. coli flagellin + N-terminal segment of the hypervariable region of H. pylori flagellin FlaA from 213 AA; useful for inserting protein segments into the chimeric flagellin (example: CF213-multiepitope-CF215)

CF215 – the C-terminal portion of chimeric fusion protein; it contains the C-terminal segment of the hypervariable region of H. pylori flagellin FlaA from 215 AA + 99 AA of C-terminal portion from E. coli flagellin; useful for inserting protein segments into the chimeric flagellin (example: CF213-multiepitope-CF215)

We also designed synthetic multiepitope, comprising the H. pylori urease B epitope, a vacuolating cytotoxin A (Vac) epitope and an adhesin A (HpaA) epitope. B-cell epitopes were designed using available 3D models or structures, which enabled more accurate prediction. Multiepitope is a synthetic gene orderd from GeneArt.

Since the protein was designed to be produced, we additionally added RGD-Hisstop tag

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 2487
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 2044
  Illegal BamHI site found at 2555
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1042
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 2838