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Composite

Part:BBa_K1744001

Designed by: Kevin Neil   Group: iGEM15_Sherbrooke   (2015-09-14)
Revision as of 17:53, 14 September 2015 by Neik1601 (Talk | contribs)

P1U8-amilCP-kanR


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1514
    Illegal NheI site found at 1537
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

This part is a positive selection system for recombineering. It contains a kanamycin resistance gene Aph(3’)-I (aminoglycoside 3’-phosphotransferase, truncated version) with its native promoter as well as amilCP, a deep blue chromoprotein. amilCP gene is under the control of a strong constitutive promoter with a strong RBS to make it more intense than previous versions. The cassette is designed to be a double positive selection marker for recombineering. Once amplified with primer bearing homology to the targeted region, the cassette can be integrated in the genome through lambda red recombination. Once integrated, the colonies should become resistant to kanamycin and become pale blue overtime. However, the blue color is actually faint and really slow to become apparent.

[edit]
Categories
//cds/reporter/chromoprotein
Parameters
color