Composite

Part:BBa_K2956000:Design

Designed by: Kelly Huang, Michelle Lam, Malini Basu, Neelam Pandya, David Sung, Steven Ionov, Kaavya Ashok, Ali Hasan   Group: iGEM19_NYU_New_York   (2019-10-16)


RBS-FNS under J23100 Promoter


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 794
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This part is a composite part consisting of BBa_J23100 promoter, BBa_B0034 ribosome binding sequence, and BBa_K2956000 coding region for flavone synthase (FNS). The FNS gene comes from Petroselinum crispum. These parts were assembled together to allow for constitutive expression of the FNS gene which converts naringenin to apigenin. When inserted into a vector plasmid and expressed in E. coli, naringenin was shown to be converted at a rate that was positively correlated to cell growth, demonstrating the efficacy of this system.

Silent mutations were introduced to BBa_K2956000 to remove illegal sites, allowing RFC[10] compatibility.

Source

The DNA sequences for the basic parts that are included in this system were obtained from the iGEM registry. The J23100 promoter, part BBa_J23100, was created by iGEM 2006 Berkeley and widely used in other iGEM projects for its high strength. The ribosome binding site, part BBa_B0034, was also chosen because it had been highly characterized. The coding region for flavone synthase, BBa_K2819001, was taken from Petroselinum crispum and codon optimized.

References