Composite

Part:BBa_M50442:Design

Designed by: Gabriella Schreiner   Group: Stanford BIOE44 - S11   (2018-04-25)


beta pinene synthase construct


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 806
    Illegal EcoRI site found at 980
    Illegal EcoRI site found at 1258
    Illegal SpeI site found at 1961
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 806
    Illegal EcoRI site found at 980
    Illegal EcoRI site found at 1258
    Illegal SpeI site found at 1961
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 806
    Illegal EcoRI site found at 980
    Illegal EcoRI site found at 1258
    Illegal BamHI site found at 1796
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 806
    Illegal EcoRI site found at 980
    Illegal EcoRI site found at 1258
    Illegal SpeI site found at 1961
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 806
    Illegal EcoRI site found at 980
    Illegal EcoRI site found at 1258
    Illegal SpeI site found at 1961
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1971
    Illegal SapI.rc site found at 1678


Design Notes

We inserted beta pinene synthase sequence following the T5 promotor and Strong RBS. DNA 2.0 added an IPTG inducible promotor to the start of the beta pinene synthase gene. We added a His tag + stop codon and T7 phage terminator.

Source

basic part BBa_M50438 sequence for beta pinene synthase, codon optimized for E. coli from Brenda sequence T5 Promoter Strong RBS His-tag + stop codon T7 Phage terminator

References

T5 Promoter: https://parts.igem.org/Part:BBa_M50075

Strong RBS: https://parts.igem.org/Part:BBa_M50080

His-tag + stop codon: https://parts.igem.org/Part:BBa_K133133

T7 Phage terminator: https://parts.igem.org/Part:BBa_M50060

"Sequence of QH6_ARTAN." Brenda The Comprehensive Enzyme Searcher. January 2018.

Maia, Marta Ferreira, and Sarah J Moore. “Plant-Based Insect Repellents: A Review of Their Efficacy, Development and Testing.” Malaria Journal 10.Suppl 1 (2011): S11. PMC. Web. 10 June 2018.

Lu, Shan et al. “Cloning and Functional Characterization of a Β-Pinene Synthase from Artemisia Annua That Shows a Circadian Pattern of Expression.” Plant Physiology 130.1 (2002): 477–486. PMC. Web. 10 May 2018.