Composite

Part:BBa_K3255001

Designed by: Mercedes Echaide, Paolo Natale, Laura R. Arriaga   Group: iGEM19_MADRID_UCM_HS   (2019-10-16)

Transcriptional unit for an enzyme of the athocyanine route synthesis BBa_K3255001 composite part is the transcriptional unit (TU) for gene F3H from Camelia sinensis, that codifies for enzyme flavanone3-hydroxylase in the route of anthocyanin synthesis. It transforms a flavonoid precursor, naringenin, in dihydrokaempferol. TU has been assembled using Golden Gate strategy in pARK 1 (ⲁ1) vector, containing BBa_K2656007 promotor, BBa_K2656009 RBS, BBa_K3255000 gene and BBa_K2656026 terminator.


Parts BBa_K2656007, BBa_K2656009 and BBa_K2656026 have been validated in this model as they have been effectively assembled by Golden Gate strategy, and work as expected as production of the enzyme flavanone 3-hydroxylase measured by the appearance of its product, dihydrokaemferol, indicates.

Schematic illustration of the assembly of a transcriptional unit (TU), composed of promoter (P), a ribosomal binding site (RBS), a gene and a terminator (TT) in a vector using the Golden Gate Assembly strategy.
Cultures exhibiting a deeper yellow in presence of increasing amounts of precursor, and the spectra of these cultures with the highest absorbance at 400nm at the maximum naringenin concentration.



Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 68
    Illegal PstI site found at 127
    Illegal PstI site found at 439
    Illegal PstI site found at 784
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 11
    Illegal NheI site found at 34
    Illegal SpeI site found at 68
    Illegal PstI site found at 127
    Illegal PstI site found at 439
    Illegal PstI site found at 784
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 68
    Illegal PstI site found at 127
    Illegal PstI site found at 439
    Illegal PstI site found at 784
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 68
    Illegal PstI site found at 127
    Illegal PstI site found at 439
    Illegal PstI site found at 784
  • 1000
    COMPATIBLE WITH RFC[1000]


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