Difference between revisions of "Part:BBa K313008"
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Please see [http://2010.igem.org/Team:UT-Tokyo/Sudoku_assay_LeakSw Terminator leak] assay page and our [http://2010.igem.org/Team:UT-Tokyo/Sudoku_experiments#Terminator_leak wiki]. | Please see [http://2010.igem.org/Team:UT-Tokyo/Sudoku_assay_LeakSw Terminator leak] assay page and our [http://2010.igem.org/Team:UT-Tokyo/Sudoku_experiments#Terminator_leak wiki]. | ||
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| + | ===verification experiments=== | ||
| + | This part was ligated with loxP-gfp-loxP sequence and transformed into E.coli. By incubating E.coli, then extracting plasmids, and sequencing them, it was proved that gfp sequence flanked by loxP sequence was removed. As a conclusion, cre generated by BBa_K313008 is able to catalyze site specific recombination reaction. | ||
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
Latest revision as of 19:34, 5 November 2010
pT7 - Cre recombinase generator
It is the generator of the Cre recombinase.
Cre is transcribed by the T7 RNA polymerase.
By this construct, we conducted recombination experiment along with "Cre recombinase assay device". (Please see here.)
This construct also serves as a positive control of "Cre generator with terminator" series as shown the table below.
| number | description | Subparts compositon | PartsID |
|---|---|---|---|
| 1 | Cre generator with single terminator | pT7-t90%-rbs-cre-dt | BBa_K313017 |
| 2 | Cre generator with double terminator | pT7-t90%-t90%-rbs-cre-dt | BBa_K313018 |
| 3 | Cre generator with weak single terminator | pT7-t80%-rbs-cre-dt | BBa_K313021 |
| 4 | Cre generator with weak double terminator | pT7-t80%-t80%-rbs-cre-dt | BBa_K313022 |
Now we are planning to conduct further research, seeking for appropriate terminator for 4C3 leak-switch.
Please see [http://2010.igem.org/Team:UT-Tokyo/Sudoku_assay_LeakSw Terminator leak] assay page and our [http://2010.igem.org/Team:UT-Tokyo/Sudoku_experiments#Terminator_leak wiki].
verification experiments
This part was ligated with loxP-gfp-loxP sequence and transformed into E.coli. By incubating E.coli, then extracting plasmids, and sequencing them, it was proved that gfp sequence flanked by loxP sequence was removed. As a conclusion, cre generated by BBa_K313008 is able to catalyze site specific recombination reaction.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 431
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 117
- 1000COMPATIBLE WITH RFC[1000]