Difference between revisions of "Part:BBa K3930004"
 
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<h3>Integration of the part <a href="https://parts.igem.org/Part:BBa_K3930000" class="pr-0" target="_blank">(BBa_K3930000)</a> into the yeast genome locus XII-1</h3>
 
<h3>Integration of the part <a href="https://parts.igem.org/Part:BBa_K3930000" class="pr-0" target="_blank">(BBa_K3930000)</a> into the yeast genome locus XII-1</h3>
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<p> The part <a href="https://parts.igem.org/Part:BBa_K3930000" class="pr-0" target="_blank">(BBa_K3930000)</a> was linearized and transformed into the <i>S.cerevisiae</i> LycoYeast strain according to the Amandine transformation protocol, with 2 µg of DNA. The construction is flanked by the part <a href="https://parts.igem.org/Part:BBa_K3930004" class="pr-0" target="_blank">(BBa_K3930004)</a> and <a href="https://parts.igem.org/Part:BBa_K3930005" class="pr-0" target="_blank">(BBa_K3930005)</a>. Figure 1 shows the electrophoresis gel of PCR on colony to check the clones (clone 3 is OK).</p>
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<p> The part <a href="https://parts.igem.org/Part:BBa_K3930000" class="pr-0" target="_blank">(BBa_K3930000)</a> was linearized and transformed into the <i>S.cerevisiae</i> LycoYeast strain according to the Amandine transformation protocol, with 2 µg of DNA. The construction is flanked by parts <a href="https://parts.igem.org/Part:BBa_K3930004" class="pr-0" target="_blank">(BBa_K3930004)</a> and <a href="https://parts.igem.org/Part:BBa_K3930005" class="pr-0" target="_blank">(BBa_K3930005)</a>. Figure 1 shows the electrophoresis gel of PCR on colony to check the clones (clone 3 is OK).</p>
 
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Latest revision as of 09:08, 17 October 2021


Up integrative sequence to target locus XII-1 of S. cerevisiae genome Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 256
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 256
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 293
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 256
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 256
  • 1000
    COMPATIBLE WITH RFC[1000]

Introduction

The sequences to target the integration locus XII-1 (Chr XII: 795787..796720) 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_K3930005) XII-1 down part in 3'.

Results

Integration of the part (BBa_K3930000) into the yeast genome locus XII-1

The part (BBa_K3930000) was linearized and transformed into the S.cerevisiae LycoYeast strain according to the Amandine transformation protocol, with 2 µg of DNA. The construction is flanked by parts (BBa_K3930004) and (BBa_K3930005). Figure 1 shows the electrophoresis gel of PCR on colony to check the clones (clone 3 is OK).



Figure 1: Integration of pFLEUR in LycoYeast at locus XII-1

pFLEUR 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 XII-1 up (BBa_K3930004) coupled with the integrative locus (BBa_K3930005) XII-1 down part is functional to target integration at the XII-1 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.