Lck-GFP
(Plasmid #61099)

• Purpose: Expression of a membrane-targeted form of eGFP
• Depositing Lab: Steven Green
• Publication: Benediktsson et al J Neurosci Methods. 2005 Jan 30;141(1):41-53.

Backbone

• Vector backbone: EGFP-C1
• Backbone manufacturer: Clontech
• Backbone size w/o insert (bp): 4731
• Modifications to backbone: This modified GFP is myristoylated and targeted to the plasma membrane (rafts). It was constructed from a parent Lck-GFPN1 construct. The parent Lck sequence was removed and replaced with a new Lck sequence containing two glycine residues at the C-term of Lck, as opposed to an isoleucine and a valine. The new Lck sequenced was prepared by PCR with BglII at the 5’ end and EcoRI at the 3’ end. The parent Lck was digested with BglII and EcoRI, removing the old Lck sequence, and insertng the new Lck using the same restriction sites.
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH10B
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Lck-GFP
• Promoter: CMV
• Tag / Fusion Protein:
  • EGFP (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BglI (unknown if destroyed)
• 3′ cloning site: EcoRI (unknown if destroyed)
• 5′ sequencing primer: EGFP-C
• 3′ sequencing primer: SV40 poly A Reverse

Resource Information

• A portion of this plasmid was derived from a plasmid made by: See publication for cloning details.
• Articles Citing this Plasmid:
  • 12 References

Terms and Licenses

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