YCplac111-KOG1 (pAN58)
(Plasmid #39209)

• Depositing Lab: Michael Hall
• Publication: Loewith et al Mol Cell. 2002 Sep;10(3):457-68.

Backbone

• Vector backbone: YCplac111
• Backbone size w/o insert (bp): 6100
• Vector type: Yeast Expression
• Selectable markers: LEU2

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: KOG1
• Species: S. cerevisiae (budding yeast)
• Mutation: L1512P
• Entrez Gene: KOG1 (a.k.a. YHR186C, LAS24)
• Promoter: KOG1
• Tag / Fusion Protein:
  • myc (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: XbaI (unknown if destroyed)
• 3′ cloning site: Asp718I (unknown if destroyed)
• 5′ sequencing primer: M13 reverse
• 3′ sequencing primer: M13 forward20

Terms and Licenses

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

Depositor Comments

Expresses 2x-myc-tagged KOG1 under control of KOG1 promoter. The two myc-tags are integrated between codons 1 and 2 of KOG1.

A 6.0 kb PCR fragment containing the KOG1 locus (−972 to +304 with respect to the ORF) was cloned into the XbaI and Asp718I sites of YCplac111; the sequence ATG ATC CCC GGG TTA ATT AAC GGT GAA CAA AAG CTA ATC TCC GAG GAA GAC TTG AAC GGT GAA CAA AAA TTA ATC TCA GAA GAA GAC TTG AAC GGA CTC GAC GGT GAA CAA encoding two myc tags was integrated between codons 1 and 2 of KOG1

Addgene's sequencing result identified a L1512P mutation in the KOG1 sequence when compared to GenBank reference sequence NP_012056.1. The depositing laboratory confirmed the plasmid is functional as described in the associated publication.