pAIP4_5HIH
(Plasmid #234205)

• Purpose: Heterologous protein expression of replicase polyprotein 1a (pp1a) (ORF1a polyprotein) in Escherichia coli
• Depositing Lab: Christopher Bahl
• Publication: AI Proteins, Inc. plasmids (unpublished)

Backbone

• Vector backbone: pET29b
• Backbone size w/o insert (bp): 5679
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH10B
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: 5HIH
• Alt name: R1A_MERS1
• Alt name: Replicase polyprotein 1a (pp1a) (ORF1a polyprotein)
• Species: Middle East respiratory syndrome-related coronavirus
• Insert Size (bp): 495
• Entrez Gene: orf1ab (a.k.a. G128_gp01)
• Promoter: T7
• Tag / Fusion Protein:
  • 10x HisTag-Smt3 (N terminal on backbone)

Cloning Information

• Cloning method: Golden Gate
• 5′ sequencing primer: T7-promoter
• 3′ sequencing primer: T7-Term

Resource Information

• Supplemental Documents:
  • pAIP4_5HIH.gb
• A portion of this plasmid was derived from a plasmid made by: Synthetic DNA

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Industry MTA

Depositor Comments

5' cloning site: BsaI (destroyed), 3' cloning site: BsaI (destroyed). Replicase polyprotein 1a (pp1a) (ORF1a polyprotein) [Cleaved into Host translation inhibitor nsp1 (nsp1) (Leader protein); Non-structural protein 2 (nsp2) (p65 homolog); Papain-like proteinase (PL-PRO) (EC 3.4.19.12) (EC 3.4.22.69) (Non-structural protein 3) (nsp3); Non-structural protein 4 (nsp4); 3C-like proteinase (3CL-PRO) (3CLp) (EC 3.4.22.-) (nsp5); Non-structural protein 6 (nsp6); Non-structural protein 7 (nsp7); Non-structural protein 8 (nsp8); Non-structural protein 9 (nsp9); Non-structural protein 10 (nsp10) (Growth factor-like peptide) (GFL); Non-structural protein 11 (nsp11)]