Part:BBa_K1025003

Thu-E Mutation Part

iGEM mut part is a plasmid constructed by 2013 iGEM Tsinghua-E team which is used for genome level in vivo high-diversity library construction.

Usage and Biology

In this vector, highly error-prone dnaQ mutant, mutD1 was cloned downstream of araBAD promoter so that we can control the mutation intensity of the target genome by the concentration of araBAD promoter’s inducer, L-arabinose in a strict manner. And the GFP downstream mutD was to test whether the mutD can be translated.

Functional Parameters

We measured the mutation increase induced by our mut part with the protocol described in our note from. To be brief, by measuring the reversion of rifampinresistance caused by mutation in genome, we accurately determined the mutation rate increase to be as big as four times compared with negative control (either bacteria without mutD or without arabinose input). The data is shown below. File:MutD-1.png

Sequence and Features

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 618
  Illegal NgoMIV site found at 693
  Illegal NgoMIV site found at 723
  Illegal NgoMIV site found at 943
  Illegal NgoMIV site found at 1061
  Illegal NgoMIV site found at 1328
  Illegal NgoMIV site found at 1523
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 1750