Composite

Part:BBa_K887004:Design

Designed by: WU, CHENG-HSUAN   Group: iGEM12_NCTU_Formosa   (2012-09-21)

Plac+alsS+ilvC+ilvD(each preceded by own zinc-finger and RBS)+Ptet+B0032+kivD+B0015


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 7718
    Illegal XhoI site found at 6049
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 6938
    Illegal AgeI site found at 3158
    Illegal AgeI site found at 4143
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2744
    Illegal BsaI site found at 5835
    Illegal BsaI site found at 6092
    Illegal BsaI.rc site found at 836
    Illegal BsaI.rc site found at 1430
    Illegal BsaI.rc site found at 3563


Design Notes

This part(BBa_K887004) contains a Plac promoter, three strong expressing RBS(B0034), three zinc fingers(zif268, PBSII, HIVC)encoded in front of three enzymes(alsS, ilvC, ilvD),37℃RBS and tetR gene and a strong terminator(J61048). The 37℃RBS and tetR gene would control the expression of the fourth enzyme(kivD).(TetR protein as a inhibitor of Ptet promoter.)

We fused zinc fingers with enzymes of isobutnol biosynthesis pathway, and make the DNA program as a scaffold for the fusion proteins to hold on. Therefore, the enzymes could bind the DNA program in order. The intermediates would be converted into the following compounds step by step. It helps enhance the efficiency of isobutanol production.

Source

It is improved form NCTU-Formosa 2011 iGEM team.

References

Atsumi, S.; T. Hanai and J.C. Liao (2008) Non-Fermentative Pathways for Synthesis of Branched-Chain Higher Alcohols as Biofuels, Nature, 451:86-89.