Coding

Part:BBa_K4188020

Designed by: Tobias Bröker   Group: iGEM22_WWU_Muenster   (2022-09-30)


modified ApL3H for the cytosolic expression

The genes listed in the table below were used by the 2022_iGEM Team Muenster to produce alpha-pinene and verbenone/verbenol. The alpha-pinene synthase (PptAPS) listed in the table stems from Pinus ponderosa and the chloroplast tag was removed. The enzyme catalyzes the reaction of geranyl diphosphate (GPP) or neryl diphosphate (NPP) to alpha-pinene via a ring closure. The enzyme limonene-3-hydroxylase (L3H) from Aureobasidium pullulans can then produce verbenone/verbenol from alpha-pinene by oxidation. All genes are MoClo (RFC1000) compatible.


Genes for the verbenone production

part number Gene length
BBa_K4188020 ApL3H 1584 bp
BBa_K4188031 PptAPS 1746 bp
BBa_K4188032 PptAPS_PTS1 1755 bp

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 352
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 126
    Illegal PstI site found at 352
    Illegal NotI site found at 64
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1217
    Illegal BglII site found at 1433
    Illegal XhoI site found at 730
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 352
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 352
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None