Part:BBa_K260015

P_FRT_dhfr. (strong promoter, translated FRT site, trimethoprim resistance)

This is the first of two parts of a FLP recombinase reporter. It has a strong promoter, a translated FRT site, and a codon-optimised gene for trimethoprim resistance (TmpR) called dhfr.

The second part of this FLP recombinase reporter is the TT_FRT_GFP BioBrick (BBa_K260016). They can both be transferred to the pCC2FosM (BBa_K260000) and pCC2FosMB (BBa_K260001) backbones, at defined loci to vary the distance between both FRT sites, of which each part has exactly one.

The position of the TT_FRT_GFP BioBrick (BBa_K260016) is always the same, whereas the position of P_FRT_dhfr (BBa_K260015) can be varied to give inter-FRT distances of 500 bp, 1 kb, 2 kb, 5 kb, 10 kb. This is achieved by starting with different backbones to contain the present BioBrick part (BBa_K260015), that have specific homology arms for recombineering-mediated transfer of P_FRT_dhfr to defined positions:

BBa_K260004: pMA-@CC2FosMB-00500bp

BBa_K260005: pMA-@CC2FosMB-01000bp

BBa_K260006: pMA-@CC2FosMB-02000bp

BBa_K260007: pMA-@CC2FosMB-05000bp

BBa_K260008: pMA-RQ-@CC2FosMB-10000bp

See BBa_K260016 for how this reporter works.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal XbaI site found at 79
  
• 12
  INCOMPATIBLE WITH RFC[12]

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

  Illegal XbaI site found at 79
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 79
  
• 1000
  COMPATIBLE WITH RFC[1000]