Coding

Part:BBa_K4188032

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


PptAPS fused to PTS1 tag for the peroxisomal 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. The gene is 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 EcoRI site found at 787
    Illegal PstI site found at 1040
    Illegal PstI site found at 1360
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 787
    Illegal PstI site found at 1040
    Illegal PstI site found at 1360
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 787
    Illegal BamHI site found at 1059
    Illegal XhoI site found at 834
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 787
    Illegal PstI site found at 1040
    Illegal PstI site found at 1360
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 787
    Illegal PstI site found at 1040
    Illegal PstI site found at 1360
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None