Difference between revisions of "Part:BBa K260015"
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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. | 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 second part of this FLP recombinase reporter is the TT_FRT_GFP BioBrick ([[Part:BBa_K260016|BBa_K260016]]). They can both be transferred to the pCC2FosM ([[Part:BBa_K260000|BBa_K260000]]) and pCC2FosMB ([[Part:BBa_K260001|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: | + | The position of the TT_FRT_GFP BioBrick ([[Part:BBa_K260016|BBa_K260016]]) is always the same, whereas the position of P_FRT_dhfr ([[Part:BBa_K260015|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|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_K260004|BBa_K260004]]: pMA-@CC2FosMB-00500bp |
− | [[Part:BBa_K260005|]]: pMA-@CC2FosMB-01000bp | + | [[Part:BBa_K260005|BBa_K260005]]: pMA-@CC2FosMB-01000bp |
− | [[Part:BBa_K260006|]]: pMA-@CC2FosMB-02000bp | + | [[Part:BBa_K260006|BBa_K260006]]: pMA-@CC2FosMB-02000bp |
− | [[Part:BBa_K260007|]]: pMA-@CC2FosMB-05000bp | + | [[Part:BBa_K260007|BBa_K260007]]: pMA-@CC2FosMB-05000bp |
− | [[Part:BBa_K260008|]]: pMA-RQ-@CC2FosMB-10000bp | + | [[Part:BBa_K260008|BBa_K260008]]: pMA-RQ-@CC2FosMB-10000bp |
− | See [[Part:BBa_K260016|]] for how this reporter works. | + | See [[Part:BBa_K260016|BBa_K260016]] for how this reporter works. |
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here |
Revision as of 15:24, 18 October 2009
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
- 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]