Part:BBa_K1486032

sfGFP + ADH1 terminator + Ura3 cassette

This part is a tagging vector for the Hog1 protein of the budding yeast S.cerevisiae. It contains the sequence of the YO superfolder GFP (optimized for yeast cells) linked to the Ura3 selection marker gene. We used it as a tag to control the expression of our gene of interest Hog1. 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 Hog1 gene and the 50 first non-coding bases after the gene, targeting this construct into the genome at the right place, fused to Hog1.

Source

It was extracted by PCR from the plasmid we ordered to addgene: Plasmid 44873 pFA6a-link-yoSuperfolderGFP-CaURA3.

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 1547
  Illegal EcoRI site found at 2478
  Illegal PstI site found at 1236
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 1547
  Illegal EcoRI site found at 2478
  Illegal PstI site found at 1236
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 1547
  Illegal EcoRI site found at 2478
  Illegal BglII site found at 1026
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 1547
  Illegal EcoRI site found at 2478
  Illegal PstI site found at 1236
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 1547
  Illegal EcoRI site found at 2478
  Illegal PstI site found at 1236
  
• 1000
  COMPATIBLE WITH RFC[1000]