Difference between revisions of "Part:BBa K3930013"
 
Line 8: Line 8:
  
 
<h2>Introduction</h2>
 
<h2>Introduction</h2>
−
<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).
+
<p>The sequence to target the locus X-3 (Chr X: 223616..224744) of <i>S. cerevisiae</i> 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 <a href="https://parts.igem.org/Part:BBa_K3930012" class="pr-0" target="_blank">(BBa_K3930012)</a> X-3 up part in 5'.</p>
 
This part is flanking the insert in 3' and must be used with the <a href="https://parts.igem.org/Part:BBa_K3930012" class="pr-0" target="_blank">(BBa_K3930012)</a> X-3 up part in 5'.</p>
 
<h2>Results</h2>
 
<h2>Results</h2>

Latest revision as of 08:59, 17 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 (Chr X: 223616..224744) 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_K3930012) and (BBa_K39300013). Figure 1 shows the electrophoresis gel of PCR on colony to check the clones.



Figure 1: Integration of pFRAMBOISE-fused 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.