pF150 pSOD1G93AAcGFP1
(Plasmid #26411)

• Purpose: contains G94A mutation in pSOD1
• Depositing Lab: Elizabeth Fisher
• Publication: Stevens et al PLoS One. 2010 . 5(3):e9541.

Backbone

• Vector backbone: pAcGFP1-N1
• Backbone manufacturer: Clontech
• Backbone size w/o insert (bp): 4700
• 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): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: SOD1
• Species: H. sapiens (human)
• Insert Size (bp): 470
• Mutation: G94A
• Entrez Gene: SOD1 (a.k.a. ALS, ALS1, HEL-S-44, IPOA, SOD, STAHP, hSod1, homodimer)
• Tag / Fusion Protein:
  • GFP (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: SalI (not destroyed)
• 5′ sequencing primer: CMV-F

Resource Information

• A portion of this plasmid was derived from a plasmid made by: human cDNA insert from cDNA clone cSOD1 from Professor Yoram Groner as described in Lieman-Hurwitzet al., 1982 and Sherman et al 1983.
• Articles Citing this Plasmid:
  • 5 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

The complete cDNA was subcloned into the vector pET28. Site directed mutagenesis changed wildtype DNA sequence for codon 93 of the cDNA from GGT to GCT, resulting in the G93A mutation in the protein and then a PCR amplified the complete cDNA with EcoRI and SalI sites on the 5’ and 3’ end respectively. This EcoRI-Sal1 0.47kb fragment was cloned into the respective EcoRI and SalI sites of pAcGFP1N1. Thus SOD1 is 5’ of GFP and SOD1 lies upstream of GFP in the SOD1-GFP fusion protein.