Difference between revisions of "Part:BBa K313008"
(→‎verification experiments)
 
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Cre is transcribed by the T7 RNA polymerase.
 
Cre is transcribed by the T7 RNA polymerase.
  
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We conducted recombination experiment using this part and Cre recombinase assay device.
+
By this construct, we conducted recombination experiment along with "Cre recombinase assay device".
−
(Please see [Part:BBa_K313009 here].)
+
(Please see [https://parts.igem.org/Part:BBa_K313009 here].)
 +
 
 +
This construct also serves as a positive control of "Cre generator with terminator" series as shown the table below.
 +
 
 +
<table class="wikitable">
 +
    <caption>Other Cre generator with terminator parts</caption>
 +
    <tr>
 +
        <th>number</th><th>description</th><th>Subparts compositon</th><th>PartsID</th>
 +
    </tr>
 +
    <tr>
 +
        <th>1</th><td>Cre generator with single terminator</td><td>pT7-t90%-rbs-cre-dt</td><td>BBa_K313017</td>
 +
    </tr>
 +
    <tr>
 +
        <th>2</th><td>Cre generator with double terminator</td><td>pT7-t90%-t90%-rbs-cre-dt</td><td>BBa_K313018</td>
 +
    </tr>
 +
    <tr>
 +
        <th>3</th><td>Cre generator with weak single terminator</td><td>pT7-t80%-rbs-cre-dt</td><td>BBa_K313021</td>
 +
    </tr>
 +
    <tr>
 +
        <th>4</th><td>Cre generator with weak double terminator</td><td>pT7-t80%-t80%-rbs-cre-dt</td><td>BBa_K313022</td>
 +
    </tr>
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</table>
  
 
Now we are planning to conduct further research, seeking for appropriate terminator for 4C3 leak-switch.  
 
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.
+
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.
  
 
<!-- 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.

Other Cre generator with terminator parts
numberdescriptionSubparts compositonPartsID
1Cre generator with single terminatorpT7-t90%-rbs-cre-dtBBa_K313017
2Cre generator with double terminatorpT7-t90%-t90%-rbs-cre-dtBBa_K313018
3Cre generator with weak single terminatorpT7-t80%-rbs-cre-dtBBa_K313021
4Cre generator with weak double terminatorpT7-t80%-t80%-rbs-cre-dtBBa_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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 431
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 117
  • 1000
    COMPATIBLE WITH RFC[1000]