Part:BBa_K515104

J23100 promoter - Antiholin

This sequence has been taken from BBa_K112808 (Berkeley 2008's kill switch cassette) and has been modified through the addition of an insulator sequence between our newly created RBS and the coding sequence. The insulator sequence has been specially developed to allow the easy switching of our parts between promoters while maintaining little homology with the vector making recombination events between the insulator sequence and the vector unlikely. We have also added a double TAA terminator sequence instead of the original single TGA sequence.

A composite part between J23100 and anti-holin (BBa_K515004). The promoter has been chosen in order to comply with our Gene Guard device specifications. It is under the influence of a strong promoter due to the lower copy number of this gene (will be integrated into the genome) in comparison to that of the Holin gene which will be in a high copy number plasmid within our system.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 276
  

This BioBrick has been sequence verified. It has also been tested for protein expression. A nice clear band of the appropriate size can be seen at the bottom of the gel indicating a strong over-expression of a small protein of the appropriate size.

Figure 1: Protein gel showing an over-expression band (higher in intensity) of a small protein. Lane 1 and 2 are control and lane 3 and 4 are the anti-holin expressing cells.(Data by Imperial College London iGEM team 2011).