Composite

Part:BBa_K3757007:Design

Designed by: Erik Zimmer   Group: iGEM21_Tuebingen   (2021-09-28)


35S Oak1


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1468
    Illegal BamHI site found at 1858
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 66


Design Notes

Codon-usage optimized for expression in Nicotiana benthamiana; composite part was cloned by Golden Gate assembly from lower-level vectors; point mutations introduced in 35S promoter to remove BpiI restriction sites.


Source

Oak1 (BBa_K3757001) from Oldenlandia affinis, sequence from GenBank: AF393825.1 MCoTI-II_loop5_His6 (BBa_K3757002) from Momordica cochinchinensis. Amino acid sequence from Cybase (http://www.cybase.org.au/index.php?page=card&table=protein&id=3) reverse transcibed into DNA sequence with codon usage optimized for Nicotiana bethaniana. c-myc tag originates from the c-myc gene, provided in Golden Gate vector. 35S promoter (BBa_K788000) and terminator (BBa_K1159307) originate from cauliflower mosaic virus, provided in Golden Gate vectors.

References