pF143 pAcGFP1 SOD1G37R
(Plasmid #26404)

• 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: G37R
• 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 37 of the cDNA from GGA to AGA (*see Addgene's note below), resulting in the G37R 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.
*Please note that the Addgene verified sequence found that codon 37 was mutated to CGT which still encodes Arginine.