Difference between revisions of "Part:BBa K3930021"
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<h2>Introduction</h2>
 
<h2>Introduction</h2>
 
<p>The sequence of the integrative locus up XII-4 of <i>S. cerevisiae</i> genome comes from the EasyClone-MarkerFree kit (Jessop-Fabre et al.,2016).
 
<p>The sequence of the integrative locus up XII-4 of <i>S. cerevisiae</i> genome comes from the EasyClone-MarkerFree kit (Jessop-Fabre et al.,2016).
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This part is flanking the insert 5', and must be used with the integrative locus (BBa_K3930022) XII-4 up part in 3'.</p>
+
This part is flanking the insert in 5', and must be used with the integrative locus (BBa_K3930022) XII-4 up part in 3'.</p>
 
<h2>Results</h2>
 
<h2>Results</h2>
  

Revision as of 14:52, 15 October 2021


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


Assembly Compatibility:
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    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 of the integrative locus up XII-4 of S. cerevisiae genome comes from the EasyClone-MarkerFree kit (Jessop-Fabre et al.,2016). This part is flanking the insert in 5', and must be used with the integrative locus (BBa_K3930022) XII-4 up part in 3'.

Results

Integration of part (BBa_K3930001) into the yeast genome

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 parts (BBa_K39300021) and (BBa_K39300022). Figure 1 shows the electrophoresis gel of colony PCR to verify integrants genotype.



Figure 1: Integration of pVIOLETTE insert in the LycoYeast genome

pVIOLETTE insert integration was checked by PCR visualised on EtBr stained agarose electrophoresis gel. A theoretical gel is presented on the right and the NEB 1 kb DNA ladder on the left (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 are functional under these 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.