pF142 pAcGFP1 SOD1 A4V
(Plasmid #26403)

• Depositing Lab: Elizabeth Fisher
• Publication: Stevens et al PLoS One. 2010 . 5(3):e9541.

Backbone

• Vector backbone: pAcGFP1-C1
• 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: A4V
• Entrez Gene: SOD1 (a.k.a. ALS, ALS1, HEL-S-44, IPOA, SOD, STAHP, hSod1, homodimer)
• Tag / Fusion Protein:
  • GFP (N 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
• 3′ sequencing primer: SV40pA-R

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.

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 4 of the cDNA from GCC to GTC, resulting in the A4V 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 pACGFPC1. Thus GFP is 5’ of the SOD1 insert and upstream in the GFP-SOD1 fusion protein.