Composite

Part:BBa_K2423000:Design

Designed by: iGEM Uppsala 2017   Group: iGEM17_Uppsala   (2017-08-18)


CaCCD2 under the control of a constitutive promoter (BBa_J23106)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1774
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 672


Design Notes

We made a complete device including promoter and RBS. We also included a 6x his tag for easy purification of the expressed protein.


Source

Originally, the group enzymes was discovered by Frusciante et al. (2014) (1) through deep transcriptome analysis. More specific information about CaCCD2 came approximately one year later Ahrazem et al. (2015) (2). The sequencing they found there has the GenBank acession number KP792756.

References

1. Frusciante S, Diretto G, Bruno M, Ferrante P, Pietrella M, Prado-Cabrero A, et al. Novel carotenoid cleavage dioxygenase catalyzes the first dedicated step in saffron crocin biosynthesis. Proc Natl Acad Sci USA. 2014 Aug 19;111(33):12246–51.

2. Ahrazem O, Rubio-Moraga A, Berman J, Capell T, Christou P, Zhu C, et al. The carotenoid cleavage dioxygenase CCD2 catalysing the synthesis of crocetin in spring crocuses and saffron is a plastidial enzyme. New Phytol. 2016 Jan 1;209(2):650–63.

3. Crocus ancyrensis carotenoid cleavage dioxygenase (CCD2) mRNA, complete cds. 2015 Nov 4; Available from: http://www.ncbi.nlm.nih.gov/nuccore/KP792756.1