Part:BBa_K1486026

sfGFP + Kanamycin resistance for yeast

Purpose of the Biobrick

This part is an integrative tagging vector for the Pbs2 protein of the budding yeast S.cerevisiae. It contains the sequence of the YO superfolder GFP (optimized for yeast cells, preceded by an 8 amino acids long linker) linked to the KanMx resistance gene. We used it as a tag to control the expression of our gene of interest Pbs2. The part is flanked in both 5' and 3' ends by 50 base pairs of sequence homologous to, respectively, the last 50 coding bases of the Pbs2 gene and the 50 first non-coding bases after the gene, targeting this construct into the genome at the right place, fused to Pbs2.

Source

It was extracted by PCR from the plasmid we ordered to addgene:44901 pFA6a-link-yoSuperfolderGFP-Kan.

References

Improved Blue, Green, and Red Fluorescent Protein Tagging Vectors for S. cerevisiae. Lee et al (PLoS One. 2013 Jul 2;8(7):e67902. doi: 10.1371/journal.pone.0067902. Print 2013. PubMed)

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 2475
  Illegal PstI site found at 1239
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 2475
  Illegal PstI site found at 1239
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 2475
  Illegal BglII site found at 1029
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 2475
  Illegal PstI site found at 1239
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 2475
  Illegal PstI site found at 1239
  
• 1000
  COMPATIBLE WITH RFC[1000]