pDS240
(Plasmid #34966)

• Purpose: Expresses Mid2(SS/TM), containing only the signal sequence and transmembrane helix, fused to GFP and LOVpep(K533∆, high-affinity epitope).
• Depositing Lab: Michael Glotzer
• Publication: Strickland et al Nat Methods. 2012 Mar 4. doi: 10.1038/nmeth.1904.

Backbone

• Vector backbone: YIPlac128
• Backbone size w/o insert (bp): 4263
• Modifications to backbone: PGAL-Mid2(SS/TM)-GFP-LOVpep-TCYC1 cassette inserted into YIPlac128
• 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: High Copy

Gene/Insert

• Gene/Insert name: Mid2(SS/TM)-GFP-LOVpep
• Alt name: Constitutively active LOVpep
• Species: S. cerevisiae (budding yeast); A. sativa
• Insert Size (bp): 2229
• Mutation: K533∆, high-affinity epitope
• Promoter: GAL1

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SacI (not destroyed)
• 3′ cloning site: SphI (destroyed during cloning)
• 5′ sequencing primer: M13 fwd
• 3′ sequencing primer: M13 rev

Resource Information

• Supplemental Documents:
  • Genbank sequence
• A portion of this plasmid was derived from a plasmid made by: GFP(S65T) CDS and other sequences from pGREG plasmids (EUROSCARF). A. sativa LOV2 CDS from Kevin Gardner, UT Southwestern Medical Center

Terms and Licenses

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

Depositor Comments

The LOV2 domain is amino acids 404-540 and differs from the wild-type A. sativa LOV2 sequence by the point mutations G528A and N534E. See Strickland et al. for details.