Composite
Ag43 RCF10

Part:BBa_K1352000:Design

Designed by: Ana-Maria Cujba, Martyna Sroka   Group: iGEM14_Aberdeen_Scotland   (2014-10-01)

Ag43 with PstI sites removed under the control of pBad/araC promoter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1945
    Illegal NgoMIV site found at 3169
    Illegal NgoMIV site found at 3679
    Illegal NgoMIV site found at 3700
    Illegal AgeI site found at 979
    Illegal AgeI site found at 2695
    Illegal AgeI site found at 3439
    Illegal AgeI site found at 3853
    Illegal AgeI site found at 4095
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961


Design Notes

We found that BBa_K759001 contained 6 additional PstI sites within the coding region of Ag43.
We therefore removed the additional PstI sites through PCR-based Site-Directed Mutagenisis, to meet the requirements iGEM Assemly Standard RCF10.

Source

Rescued from BioBrick BBa_K1352000 and subjected to PCR-based Site Directed Mutagenisis.

References