pLEX307-SARS-CoV-2-3CL-C145A
(Plasmid #160279)

• Purpose: Allows for constituitive mammalian expression of the SARS-CoV-2 3CL protease with C145A mutation via transfection or lentivirus
• Depositing Lab: Alejandro Chavez
• Publication: Resnick et al J Virol. 2021 Jun 24;95(14):e0237420. doi: 10.1128/JVI.02374-20. Epub 2021 Jun 24.

Backbone

• Vector backbone: pLEX_307
• Vector type: Mammalian Expression, Lentiviral
• Selectable markers: Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): NEB Stable ; NEB 10Beta
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: 3CL Protease
• Species: SARS-CoV-2
• Insert Size (bp): 924
• Mutation: C145A
• Promoter: EF1a

Cloning Information

• Cloning method: Gateway Cloning
• 5′ sequencing primer: Unknown

Resource Information

• Supplemental Documents:
  • pLEX307-SARS-CoV-2-3CL-C145A.gb

Terms and Licenses

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

Depositor Comments

This plasmid is stored and distributed in NEB Stable. It can also be used to transform NEB 10Beta.

The authors have noticed that there are high rates of recombination in these generated vectors. All users are advised to sequence the vectors and run them on a gel to ensure they show the expected full length insert and plasmid size. In cases where recombination has occurred it is typically between the viral LTR elements in the vector although recombination between the gateway cloning sites has also been observed.

Please visit https://doi.org/10.1101/2020.08.29.272864 for bioRxiv preprint.