pAIP4_6WEY
(Plasmid #234234)

• Purpose: Heterologous protein expression of replicase polyprotein 1ab (pp1ab) (ORF1ab 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• Publication: AI Proteins, Inc. plasmids (unpublished)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: 6WEY
• Alt name: R1AB_SARS2
• Alt name: Replicase polyprotein 1ab (pp1ab) (ORF1ab polyprotein)
• Species: Severe acute respiratory syndrome coronavirus 2
• Insert Size (bp): 516
• Entrez Gene: ORF1ab (a.k.a. GU280_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_6WEY.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 1ab (pp1ab) (ORF1ab polyprotein) [Cleaved into Host translation inhibitor nsp1 (Leader protein) (Non-structural protein 1) (nsp1); Non-structural protein 2 (nsp2) (p65 homolog); Non-structural protein 3 (nsp3) (EC 3.4.19.12) (EC 3.4.22.-) (PL2-PRO) (Papain-like protease) (Papain-like proteinase) (PL-PRO); Non-structural protein 4 (nsp4); 3C-like proteinase (3CL-PRO) (3CLp) (EC 3.4.22.69) (Main protease) (Mpro) (Non-structural protein 5) (nsp5) (SARS coronavirus main proteinase); 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); RNA-directed RNA polymerase (Pol) (RdRp) (EC 2.7.7.48) (Non-structural protein 12) (nsp12); Helicase (Hel) (EC 3.6.4.12) (EC 3.6.4.13) (Non-structural protein 13) (nsp13); Proofreading exoribonuclease (ExoN) (EC 3.1.13.-) (Guanine-N7 methyltransferase) (Non-structural protein 14) (nsp14); Uridylate-specific endoribonuclease (EC 3.1.-.-) (NendoU) (Non-structural protein 15) (nsp15); 2'-O-methyltransferase (EC 2.1.1.-) (Non-structural protein 16) (nsp16)]