Part:BBa_K3930022

Down 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
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Introduction

The sequences 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 3' and must be used with the (BBa_K3930021) XII-4 up part in 5'.

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 insert is flanked by parts (BBa_K3930021) 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 at locus XII-4 was checked by PCR and the result was visualized 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 sequence XII-4 down (BBa_K3930022) coupled with the XII-4 up part (BBa_K3930021) is functional to target integration at the XII-4 locus 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.