pET16b-PCNA
(Plasmid #134898)

• Purpose: Bacterial expression and purification of human PCNA
• Depositing Lab: Andrew Deans
• Publication: Tan et al PLoS One. 2020 Feb 24;15(2):e0229000. doi: 10.1371/journal.pone.0229000. eCollection 2020.

Backbone

• Vector backbone: pET16b
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5711
• Total vector size (bp): 6440
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: PCNA
• Alt name: Proliferating Cell Nuclear Antigen
• Species: H. sapiens (human)
• Insert Size (bp): 783
• Mutation: fully synthetic, codon optimized for bacterial expression
• Entrez Gene: PCNA (a.k.a. ATLD2)
• Promoter: T7
• Tag / Fusion Protein:
  • untagged

Cloning Information

• Cloning method: Ligation Independent Cloning
• 5′ sequencing primer: T7 promoter
• 3′ sequencing primer: T7 terminator

Resource Information

• Supplemental Documents:
  • pET_16b_PCNA.gb

Terms and Licenses

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

Depositor Comments

This is a codon optimized untagged PCNA cDNA expression vector. N- or C-terminal tags on PCNA inhibit its trimerization, so purification is instead via several steps of anion exchange and size exclusion chromatography. Protein purification protocol is as in Tan et al. Plos One 2020