pJRL2 PHO5(UASm1)pr VYFP (EB#1549)
(Plasmid #14841)

• Depositing Lab: Erin O'Shea
• Publication: Raser et al Science. 2004 Jun 18. 304(5678):1811-4.

Backbone

• Vector backbone: pJRL2
• Vector type: Yeast Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: PHO5 promoter UASm1
• Species: S. cerevisiae (budding yeast)
• Mutation: UASm1 (AAGCTT was substituted for CACGTT). Uninducible promoter. See Steger DJ et al Science 299(5603)114-116.
• Entrez Gene: PHO5 (a.k.a. YBR093C)
• Tag / Fusion Protein:
  • VYFP (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: n/a

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

pJRL2 is a yeast chromosomal integration vector that replaces the LEU2 chromosomal locus by homologous recombination. pJRL2 was constructed by insertion of the 200 bp upstream of the LEU2 start codon, an Asc I restriction enzyme site, the 200 bp downstream of the LEU2 stop codon and a 5.5 kb hisG::URA3::kanR::hisG cassette into pBluescript.

The contruct contains 1000 bp of promoter sequence, from the nucleotide immediately upstream of the start codon, followed by six base-pairs of idealized Kozak sequence AACAAA for optimal translational initiation, the coding sequence of the appropriate fluorophore (yECFP or yVYFP) and the 500 bp 3’ to the stop codon of ACT1 in the pJRL2 integrating vector.