Lyn11-CIBN-GFP
(Plasmid #79572)

• Purpose: expression of plasma membrane signal from Lyn11 fused to CIBN and tagged with GFP
• Depositing Labs: Pietro De Camilli, Olof Idevall-Hagren
• Publication: Idevall-Hagren et al Proc Natl Acad Sci U S A. 2012 Aug 28;109(35):E2316-23. doi: 10.1073/pnas.1211305109. Epub 2012 Jul 30.

Backbone

• Vector backbone: pEGFP-N1
• Vector type: Mammalian Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Lyn11
• Alt name: 11 N-terminal amino acid residues of Lyn kinase
• Species: H. sapiens (human)
• Mutation: G8R (please see depositor comments below)
• Promoter: CMV
• Tags / Fusion Proteins:
  • CIBN (CRY2-binding domain) (C terminal on backbone)
  • EGFP (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NheI (not destroyed)
• 3′ cloning site: AgeI (not destroyed)
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: EGFP-R

Terms and Licenses

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

Depositor Comments

CIBN was fused N-terminally to GFP in pEGFP-N1 using Nhe1/Age1. Complementary oligonucleotides encoding Lyn11 with flanking Nhe1-sites subsequently were annealed and ligated N-terminally to CIBN. Lipidation for plasma membrane attachment occurs at amino acids 2 and 3. The G8R mutations stabilise the protein at the plasma membrane.