Coding

Part:BBa_K346007:Design

Designed by: miao jing   Group: iGEM10_Peking   (2010-10-15)
Revision as of 05:08, 27 October 2010 by Starshine (Talk | contribs) (→‎Design Notes)

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Antigen 43


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 709
    Illegal NgoMIV site found at 1933
    Illegal NgoMIV site found at 2443
    Illegal NgoMIV site found at 2464
    Illegal AgeI site found at 1459
    Illegal AgeI site found at 2203
    Illegal AgeI site found at 2617
    Illegal AgeI site found at 2859
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Since the original antigen 43's RBS is regulated by intracellular factors such as the oxidative particles, and in our bioabsorbant kit we need to express some proteins such as DsbA which can affect the oxidative condition in the cell, it is unconvenient for us to use antigen 43. We therefore replace the RBS by a standard RBS B0034 which is not regulated by so many factors in the cell. By this design, we are able to express antigen 43 without considering the condition within the cell.

By using nest PCR, we were able to get the coding region of antigen 43 by two step PCR. We designed the primer using Primer Premier 5.0 and add standart digestion sites into them.

Source

K12 strain of E.Coli

References