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 ([[Part:BBa_K260016|]]). They can both be transferred to the pCC2FosM ([[Part:BBa_K260000|]]) and pCC2FosMB ([[Part: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 ([[Part:BBa_K260016|]]) is always the same, whereas the position of P_FRT_dhfr ([[Part: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 ([[Part:BBa_K260015|]]), that have specific homology arms for recombineering-mediated transfer of P_FRT_dhfr to defined positions:
[[Part:BBa_K260004|]]: pMA-@CC2FosMB-00500bp
[[Part:BBa_K260005|]]: pMA-@CC2FosMB-01000bp
[[Part:BBa_K260006|]]: pMA-@CC2FosMB-02000bp
[[Part:BBa_K260007|]]: pMA-@CC2FosMB-05000bp
[[Part:BBa_K260008|]]: pMA-RQ-@CC2FosMB-10000bp
See [[Part:BBa_K260016|]] for how this reporter works.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 79
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 79
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 79
- 1000COMPATIBLE WITH RFC[1000]
None |