Part:BBa_K3930021

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 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 by 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.