pGEM-GFP-URA3-GFP
(Plasmid #72606)

• Purpose: Template for creating a C-terminal GFP tag AND an internal GFP tag from a single transfection of yeast
• Depositing Lab: Bruce Granger
• Publication: Granger et al PLoS One. 2018 Jan 12;13(1):e0191194. doi: 10.1371/journal.pone.0191194. eCollection 2018.

Backbone

• Vector backbone: pGEM-5Zf(+) [cut with EcoR V and T-tailed to give pGEM-T]
• Backbone manufacturer: Promega
• Backbone size w/o insert (bp): 2964
• Total vector size (bp): 5300
• Modifications to backbone: Part of the pGEM multiple cloning site (between the EcoR V and Apa I sites) was replaced with inserts
• Selectable markers: URA3

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Submitted plasmid was grown in Lucigen E. coli strain "E. cloni 10G"
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: GFP (full)
• Alt name: Yeast enhanced green fluorescent protein 3
• Species: Aequorea victoria
• Insert Size (bp): 717
• Mutation: S65G, S72A
• GenBank ID: Accession Number U73901

Gene/Insert 2

• Gene/Insert name: URA3
• Alt name: OMP decarboxylase
• Species: Candida albicans strain 3153A
• Insert Size (bp): 1383
• Mutation: Includes full 819 bp coding sequence of URA3, 435 upstream bases, and 129 downstream bases

Gene/Insert 3

• Gene/Insert name: GFP (partial)
• Species: Aequorea victoria
• Insert Size (bp): 199
• Mutation: Includes nucleotides 501-699 of the 717 nucleotide coding sequence of GFP

Gene/Insert 4

• Gene/Insert name: Optional Linker
• Species: Candida albicans
• Insert Size (bp): 24
• Mutation: This is a segment of the YWP1 gene that encodes SSASPSGS.

Resource Information

• Supplemental Documents:
  • Addgene 72606 pGEM-GFP-URA3-GFP updated Jan 2018.pdf
• A portion of this plasmid was derived from a plasmid made by: The yeast enhanced green fluorescent protein 3 ("GFP") portion of this plasmid was obtained from Brendan Cormack (Cormack, Bertram, Egerton, Gow, Falkow & Brown, Microbiology 143: 303-311, 1997); it has the S65G and S72A mutations, is codon-optimized for Candida albicans, and is similar to GenBank Accession Number U73901.
• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

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