pET28b-His-hCE
(Plasmid #112710)

• Purpose: Expresses N-terminally His-tagged human CE/RNGTT in bacterial cells
• Depositing Lab: Daniel Schoenberg
• Publication: Trotman et al Nucleic Acids Res. 2017 Oct 13;45(18):10726-10739. doi: 10.1093/nar/gkx801.

Backbone

• Vector backbone: pET28b
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5368
• Total vector size (bp): 7221
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Grow in BL21(DE3) for protein overexpression
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: RNGTT
• Alt name: CE, hCE
• Alt name: Capping enzyme
• Alt name: RNA guanylyltransferase and 5'-phosphatase
• Species: H. sapiens (human)
• Insert Size (bp): 1854
• Mutation: R594H
• Entrez Gene: RNGTT (a.k.a. CAP1A, HCE, HCE1, hCAP)
• Promoter: T7
• Tags / Fusion Proteins:
  • 6xHis (N terminal on backbone)
  • Stop codons placed after RNGTT sequence to prevent C-terminal 6xHis tag (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NheI (not destroyed)
• 3′ cloning site: NheI (destroyed during cloning)
• 5′ sequencing primer: T7 promoter (TAATACGACTCACTATAGG)
• 3′ sequencing primer: T7 terminator (GCTAGTTATTGCTCAGCGG)

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Source of RNGTT sequence: pET21a-hCE from David Price, University of Iowa

Terms and Licenses

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