Part:BBa_M50063:Design
Ara h2 Production Device
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 212
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 212
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 376
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 212
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 212
- 1000COMPATIBLE 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).