Difference between revisions of "Part:BBa M50063:Design"

 
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===Design Notes===
 
===Design Notes===
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We had to create a tagging mechanism and secretion system for this sequence.  
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This DNA devices has 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. This device was synthesized into a plasmid containing a kanamycin resistance cassette by DNA 2.0.
  
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===Source===
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Plasmid name: pMW_IJ_export
  
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DNA2.0 Gene #: PD441-gIII
  
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===Source===
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Organism: E. coli
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Device type: Actuator
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Glycerol stock barcode: 0133027184
  
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This part comes from the Ara h2 sequence in the peanut.
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Box label: BioE44 S17
  
 
===References===
 
===References===
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Glenting, J. et al. Production of Recombinant Peanut Allergen Ara h 2 using Lactococcus lactis. Microb. Cell Factories 6, 28 (2007).

Latest revision as of 06:19, 12 June 2017


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 has 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. This device was synthesized into a plasmid 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).