Intermediate

Part:BBa_K260015

Designed by: Kaj Bernhardt   Group: iGEM09_BIOTEC_Dresden   (2009-10-18)
Revision as of 08:55, 18 October 2009 by Registry (Talk | contribs)

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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


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]


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Categories
Parameters
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