Device

Part:BBa_M50063:Design

Designed by: Isaac Justice   Group: Stanford BIOE44 - S11   (2017-04-26)
Revision as of 06:10, 12 June 2017 by Ijustice (Talk | contribs) (→‎Source)


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

Plasmid name: pMW_IJ_export DNA2.0 Gene #: PD441-gIII Organism: E. coli Device type: Actuator Glycerol stock barcode: 0133027184 Box label: BioE44 S17

References

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