Part:BBa_M50063:Design

Ara h2 Production Device

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 212
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 212
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 376
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 212
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 212
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

This DNA devices had six parts: An IPTG inducible inducible T5 promoter sourced from the iGEM registry of parts; a strong ribosomal site sourced from the iGEM registry of parts; the Ara h2 gene taken from the Ara h2 protein sequence in the paper Production of peanut antigen in L.lactis, Glenting, et al., reverse translated from protein sequence to a genetic sequence, and codon optimized for the E. coli organism; a 6x histidine tag attached to the end of the Ara h2 sequence for identification with a western blot sourced from DNA 2.0; and a T5 terminator sourced from DNA 2.0; a giii secretion tag sourced from DNA 2.0 and added directly before the Ara h2 sequence to hopefully aid in Ara h2 secretion. The two devices were synthesized into plasmids containing a kanamycin resistance cassette by DNA 2.0.

Source

Glenting, J. et al. Production of Recombinant Peanut Allergen Ara h 2 using Lactococcus lactis. Microb. Cell Factories 6, 28 (2007).

References

Glenting, J. et al. Production of Recombinant Peanut Allergen Ara h 2 using Lactococcus lactis. Microb. Cell Factories 6, 28 (2007).