Difference between revisions of "Part:BBa K3930013"
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<h2>Introduction</h2>
 
<h2>Introduction</h2>
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<p>Integrative locus down X-3 of <i>S. cerevisiae</i> genome comes from the EasyClone-MarkerFree kit (Jessop-Fabre et al.,2016).
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<p>The sequence to target the locus X-3 of <i>S. cerevisiae</i> genome come from the EasyClone-MarkerFree kit (Jessop-Fabre et al., 2016).
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This part needs to flank the insertion in 3', and must be used with the integrative locus (BBa_K3930012) X-3 up part in 5'.</p>
+
This part is flanking the insert in 3' and must be used with the (BBa_K3930012) X-3 up part in 5'.</p>
 
<h2>Results</h2>
 
<h2>Results</h2>
  
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<h3>Integration of the part (BBa_K3930001) into the yeast genome</h3>
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<h3>Integration of the part (BBa_K3930001) into the yeast genome at locus X-3</h3>
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<p> The part (BBa_K3930001) was linearized and transformed into the <i>S.cerevisiae</i> LycoYeast strain according to the Takara yeast transformation protocol, with 5 µg of DNA. The construction is flanked by the part (BBa_K39300012) and (BBa_K39300013). Figure 1 shows the electrophoresis gel of PCR on colony to verify clones.</p>
+
<p> The part (BBa_K3930001) was linearized and transformed into the <i>S.cerevisiae</i> LycoYeast strain according to the Takara yeast transformation protocol, with 5 µg of DNA. The construction is flanked by the part (BBa_K39300012) and (BBa_K39300013). Figure 1 shows the electrophoresis gel of PCR on colony to check the clones.</p>
 
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                     <a href="https://2021.igem.org/wiki/images/b/bb/T--Toulouse_INSA-UPS--2021_fig28genotyping.png" class="internal" title="Enlarge"></a>
 
                     <a href="https://2021.igem.org/wiki/images/b/bb/T--Toulouse_INSA-UPS--2021_fig28genotyping.png" class="internal" title="Enlarge"></a>
 
                 </div>
 
                 </div>
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                 <b>Figure 1: </b> <b> Integration of pFRAMBOISE in the LycoYeast genome</b>
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                 <b>Figure 1: </b> <b> Integration of pFRAMBOISE in the LycoYeast genome at locus X-3</b>
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                 <p>pFRAMBOISE was checked with agarose electrophoresis gel and revealed with EtBr. A theoretical gel is presented on the right of each gel and the NEB 1 kb DNA ladder on the left (note that a different ladder is presented on the theoretical gel)</p>
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                 <p>pFRAMBOISE was checked with agarose electrophoresis gel and revealed with EtBr. A theoretical gel is presented on the right of each gel (note that a different ladder is presented on the theoretical gel)</p>
 
             </div>
 
             </div>
 
         </div>
 
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</div>
 
</div>
 
<br>
 
<br>
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<p><b>The Integrative locus X-3 down (BBa_K3930013) coupled with the integrative locus (BBa_K3930012) X-3 up part work under those experimental conditions.</b><p>
+
<p><b>The Integrative locus X-3 down (BBa_K3930013) coupled with the integrative locus (BBa_K3930012) X-3 up part is functional to target integration at the X-3 locus under those experimental conditions.</b><p>
 
<br>
 
<br>
 
<h2>References</h2>
 
<h2>References</h2>

Revision as of 18:34, 15 October 2021


Down integrative sequence to target locus X-3 of S. cerevisiae genome Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 71
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 71
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 71
    Illegal XhoI site found at 378
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 71
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 71
  • 1000
    COMPATIBLE WITH RFC[1000]

Introduction

The sequence to target the locus X-3 of S. cerevisiae genome come from the EasyClone-MarkerFree kit (Jessop-Fabre et al., 2016). This part is flanking the insert in 3' and must be used with the (BBa_K3930012) X-3 up part in 5'.

Results

Integration of the part (BBa_K3930001) into the yeast genome at locus X-3

The part (BBa_K3930001) was linearized and transformed into the S.cerevisiae LycoYeast strain according to the Takara yeast transformation protocol, with 5 µg of DNA. The construction is flanked by the part (BBa_K39300012) and (BBa_K39300013). Figure 1 shows the electrophoresis gel of PCR on colony to check the clones.



Figure 1: Integration of pFRAMBOISE in the LycoYeast genome at locus X-3

pFRAMBOISE was checked with agarose electrophoresis gel and revealed with EtBr. A theoretical gel is presented on the right of each gel (note that a different ladder is presented on the theoretical gel)


The Integrative locus X-3 down (BBa_K3930013) coupled with the integrative locus (BBa_K3930012) X-3 up part is functional to target integration at the X-3 locus under those experimental conditions.


References

  1. Chen X, Shukal S, Zhang C. 2019. Integrating Enzyme and Metabolic Engineering Tools for Enhanced α-Ionone Production. J Agric Food Chem. 67(49):13451–13459. doi:10.1021/acs.jafc.9b00860.
    2.