Part:BBa_K1744001

P1U8-amilCP-kanR

Part Description

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.

Sequence and Features

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]