pET28a-PACT
(Plasmid #50352)

• Purpose: bacterial expression of PACT
• Depositing Lab: Kumiko Ui-Tei
• Publication: Takahashi et al PLoS One. 2013 May 2;8(5):e63434. doi: 10.1371/journal.pone.0063434. Print 2013.

Backbone

• Vector backbone: pET28a
• Backbone manufacturer: Novagen
• Total vector size (bp): 6349
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: PACT
• Alt name: PRKRA
• Alt name: protein kinase, interferon-inducible double stranded RNA dependent activator
• Species: H. sapiens (human)
• Entrez Gene: PRKRA (a.k.a. DYT16, HSD14, PACT, RAX)
• Promoter: T7
• Tags / Fusion Proteins:
  • His (N terminal on backbone)
  • Myc (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: SalI (not destroyed)
• 5′ sequencing primer: T7

Terms and Licenses

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

Depositor Comments

The PACT CDS was amplified from a cDNA mixture of total RNA extracted from human HeLa cells by PCR using primers with restriction enzyme sites of BamHI and SalI. The amplified fragment was digested with same enzyme sites and cloned into pET-28a vector amplified using primers with same enzyme sites.