Difference between revisions of "Part:BBa K3930021"
 
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<h2>Introduction</h2>
 
<h2>Introduction</h2>
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<p>Integrative locus up XII-4 of S. cerevisiae genome comes from the EasyClone-MarkerFree kit (Jessop-Fabre et al.,2016).
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<p>The sequence to target the locus XII-4 (Chr XII:830227..831248) 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 5', and must be used with the integrative locus (BBa_K3930022) XII-4 up part in 3'.</p>
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This part is flanking the insert in 5' and must be used with the <a href="https://parts.igem.org/Part:BBa_K3930022" class="pr-0" target="_blank">(BBa_K3930022)</a> XII-4 up part in 3'.</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 part <a href="https://parts.igem.org/Part:BBa_K3930003" class="pr-0" target="_blank">(BBa_K3930003)</a> into the yeast genome at locus XII-4</h3>
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<p> The part (BBa_K3930003) 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_K39300021) and (BBa_K39300022). Figure 1 shows the electrophoresis gel of PCR on colony to verify clones.</p>
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<p> The part <a href="https://parts.igem.org/Part:BBa_K3930003" class="pr-0" target="_blank">(BBa_K3930003)</a> 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 parts (BBa_K39300021) and <a href="https://parts.igem.org/Part:BBa_K3930022" class="pr-0" target="_blank">(BBa_K3930022)</a>. Figure 1 shows the electrophoresis gel of colony PCR to verify integrants genotype (clone 2 is OK).</p>
 
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                     <a href="https://2021.igem.org/wiki/images/a/a9/T--Toulouse_INSA-UPS--2021_fig23.violette.png" class="internal" title="Enlarge"></a>
 
                     <a href="https://2021.igem.org/wiki/images/a/a9/T--Toulouse_INSA-UPS--2021_fig23.violette.png" class="internal" title="Enlarge"></a>
 
                 </div>
 
                 </div>
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                 <b>Figure 1: </b> <b> Integration of pVIOLETTE in the LycoYeast genome</b>
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                 <b>Figure 1: </b> <b> Integration of pVIOLETTE insert in the LycoYeast genome at locus XII-4</b>
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                 <p>pVIOLETTE 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>pVIOLETTE insert integration was checked by PCR visualised on EtBr stained agarose electrophoresis gel. A theoretical gel is presented on the right (note that a different ladder is presented on the theoretical gel)</p>
 
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<p><b>The Integrative locus XII-4 up (BBa_K3930021) coupled with the integrative locus (BBa_K3930022) XII-4 down part work under those experimental conditions.</b><p>
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<p><b>The integrative locus XII-4 up (BBa_K3930021) coupled with the integrative locus <a href="https://parts.igem.org/Part:BBa_K3930022" class="pr-0" target="_blank">(BBa_K3930022)</a> XII-4 down part is functional to target integration at the XII-4 locus under those experimental conditions.</b><p>
 
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<h2>References</h2>
 
<h2>References</h2>

Latest revision as of 21:22, 16 October 2021


Up integrative sequence to target locus XII-4 of Saccharomyces cerevisiae genome Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 73
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Introduction

The sequence to target the locus XII-4 (Chr XII:830227..831248) of S. cerevisiae genome come from the EasyClone-MarkerFree kit (Jessop-Fabre et al., 2016). This part is flanking the insert in 5' and must be used with the (BBa_K3930022) XII-4 up part in 3'.

Results

Integration of part (BBa_K3930003) into the yeast genome at locus XII-4

The part (BBa_K3930003) 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 parts (BBa_K39300021) and (BBa_K3930022). Figure 1 shows the electrophoresis gel of colony PCR to verify integrants genotype (clone 2 is OK).



Figure 1: Integration of pVIOLETTE insert in the LycoYeast genome at locus XII-4

pVIOLETTE insert integration was checked by PCR visualised on EtBr stained agarose electrophoresis gel. A theoretical gel is presented on the right (note that a different ladder is presented on the theoretical gel)


The integrative locus XII-4 up (BBa_K3930021) coupled with the integrative locus (BBa_K3930022) XII-4 down part is functional to target integration at the XII-4 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.