Composite

Part:BBa_K2934008:Design

Designed by: Nir Litver, Shira Levi, Asaf Licht   Group: iGEM19_Technion-Israel   (2019-10-11)
Revision as of 13:20, 18 October 2019 by NirLitver (Talk | contribs) (→‎Design Notes)


pKatA-LacI-pLac-Glucose Oxidase-Histag for B. subtilis


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1268
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1564
    Illegal BsaI site found at 1895
    Illegal BsaI.rc site found at 1841
    Illegal BsaI.rc site found at 2666


Design Notes

In our lab, we devided this part into two DNA fragments, with 40 non-coding DNA spacer segment between them (that we used as a homologic region to fuse both DNA fragments via Gibson Assembly). The spacer was located between 5' UTR and the following promoter.

Primers for isolation of the gene (with RFC[10] suffix and prefix):

fwd: 5'-GGCCGCTTCTAGAGATAACTATTTT-3'

rev: 5'-GTATTAGTGGTGATGATGGTGATGTC-3'

Source

This part is made up of several basic parts: pKatA - BBa_K2934002 3' UTR - BBa_K2934007 LacI - BBa_K143033 5' UTR - BBa_K2934003 pLac - BBa_K143015 GOx with secretory signal peptide AmyE - BBa_K2934004


References