pAAV-UbC-eGFP-F
(Plasmid #71545)

• Purpose: Expresses a membrane bound eGFP, under the UbC promoter. This promoter expresses transgenes in neurons at higher levels than plasmids with the CMV or β-actin-CMV promoters.
• Depositing Lab: Pantelis Tsoulfas
• Publication: pAAV-UbC-eGFP-F (unpublished)

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Backbone

• Vector backbone: pAAV-SIBR-anti_I
• Backbone manufacturer: Chung K-H et al., Polycistronic RNA polymerase II expression vectors for RNA interference based on BIC/miR-155. NAR 2006 vol 24
• Backbone size w/o insert (bp): 5235
• Total vector size (bp): 6048
• Modifications to backbone: The anti-luciferase cassette was deleted and eGFP was replaced with eGFP that has in the COOH terminus a 20- amino-acid farnesylation signal from c-Ha-Ras. The gene was synthesized for optimal mammalian cell expression
• Vector type: Mammalian Expression, AAV

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: eGFP-F
• Alt name: membrane bound eGFP
• Species: Synthetic
• Insert Size (bp): 813
• GenBank ID: AEO31555.1 JN255744.1
• Promoter: UbC
• Tag / Fusion Protein:
  • contains 20- amino-acid farnesylation signal from c-Ha-Ras and a 4 aminoacid linker (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: common sequencing primers

Resource Information

• Supplemental Documents:
  • Insert sequence

Terms and Licenses

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