pNIC-ZB-SPRTN-E112A-FL
(Plasmid #110218)

• Purpose: Expression in E. coli of full-length SPRTN protein carrying E112A mutation, a catalytically-dead mutant, with His and ZB tags at N-terminus.
• Depositing Lab: Kristijan Ramadan
• Publication: Vaz et al Mol Cell. 2016 Nov 17;64(4):704-719. doi: 10.1016/j.molcel.2016.09.032. Epub 2016 Oct 27.

Backbone

• Vector backbone: pNIC-ZB
• Backbone manufacturer: Opher Gileadi, Addgene plasmid #26107
• Modifications to backbone: vector treated with BsaI, then with T4 DNA polymerase in presence of dGTP
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Growth Strain Other: Rosetta DE3
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: SPRTN
• Alt name: C1orf124
• Alt name: DVC1
• Species: H. sapiens (human)
• Insert Size (bp): 1494
• Mutation: E112A, P296L (see depositor comments below)
• Entrez Gene: SPRTN (a.k.a. C1orf124, DVC1, PRO4323, spartan)
• Tags / Fusion Proteins:
  • His6 (N terminal on insert)
  • ZB (N terminal on insert)

Cloning Information

• Cloning method: Ligation Independent Cloning
• 5′ cloning site: NdeI (not destroyed)
• 3′ cloning site: unknown (unknown if destroyed)
• 5′ sequencing primer: unknown

Terms and Licenses

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

Depositor Comments

According to the large datasets deposited in various databses like: dbSNP, 1000 Genomes, the ExAC and the gnomAD browser, the variant c.887C>T, p.Pro296Leu is found roughly in 50% of general world population. Moreover, according to the large-scale sequencing studies involving more than 135 000 healthy individuals deposited in The Genome Aggregation Database (gnomAD), it is more prevalent than the previously assigned reference allele in the European population, with a minor allele frequency of 0.69: http://gnomad.broadinstitute.org/variant/1-231488524-C-T
Taken this into account, this variant (P296L) is s actually the accurate reference (wild-type) allele, and thus this variant can not be the cause of any Mendelian disease. This is the reason why all our vectors bear this variant.