Pdonr223 sars cov 2 nsp14

Manufactured by Addgene

PDONR223 SARS-CoV-2 Nsp14 is a plasmid that contains the coding sequence for the SARS-CoV-2 non-structural protein 14 (Nsp14). Nsp14 is an exoribonuclease involved in viral RNA proofreading and repair. This plasmid can be used for the expression and purification of the SARS-CoV-2 Nsp14 protein.

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Lab products found in correlation

Plex 307, by Addgene (1 mentions) Gateway lr clonase 2 enzyme mix, by Thermo Fisher Scientific (1 mentions) Pezy flag, by Addgene (1 mentions) Nsp14, by Addgene (1 mentions) Mission trc3 human lentiorf library, by Merck Group (1 mentions) Lr gateway technology, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

PDONR223 (6 citations) NSP14 (3 citations)

2 protocols using pdonr223 sars cov 2 nsp14

SARS-CoV-2 Nsp Protein Expression Vectors

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pLEX-IMPDH2-V5 vector was picked from the MISSION TRC3 human LentiORF library from Sigma-Aldrich. The pcDNA-FLAG-V5-Nsp10/14/16 vectors were constructed from pDONR223 SARS-CoV-2 Nsp10 (Cat. # 141264, Addgene), Nsp14 (Cat. # 141267, Addgene), and Nsp16 (Cat. # 141269, Addgene) vectors to the pcDNA3.1–3xFLAG-V5-ccdB (Cat. # 87064, Addgene) destination vector using Gateway^™ LR Clonase^™ II Enzyme Mix (Cat. # 11791020, Invitrogen). pEZY-FLAG-Nsp14 vector was constructed from pDONR223 SARS-CoV-2 Nsp14 vector to the pEZY-FLAG (Cat # 18700, Addgene) destined vector. The pLEX-FLAG-V5 vector was constructed by cloning the FLAG sequence to the pLEX-307 (Cat # 41392, Addgene) vector. The pNF-κB-luciferase vector (PRDII₄–luc in the pGL3 vector) was the gift from Dr. Jacob Yount’s lab (61 (link)). The pIRES-luciferase vector (Cat. # 219092) was acquired from Agilent Technologies. The pRL-TK Renilla Luciferase vector (Cat. # AF025846) was purchased from Promega.

Li T., Kenney A.D., Liu H., Fiches G.N., Zhou D., Biswas A., Que J., Santoso N., Yount J.S, & Zhu J. (2021). SARS-CoV-2 Nsp14 activates NF-κB signaling and induces IL-8 upregulation. bioRxiv.

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SARS-CoV-2 NSP10 and NSP14 Cloning

Cited 1 time

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pDONR223 SARS-CoV-2 NSP10 (Addgene; plasmid no. 141264; http://n2t.net/addgene:141264; Research Resource Identifier: Addgene_141264) and pDONR223 SARS-CoV-2 NSP14 (Addgene; plasmid no. 141267; http://n2t.net/addgene:1412647; Research Resource Identifier: Addgene_141267); both were a kind gift from Fritz Roth (34 (link)). Using Gateway LR technology (Invitrogen), NSP10 and NSP14 were subcloned individually into gateway competent expression vector pDEST527 (N-terminal His-tags NSP10 and NSP14) for bacterial expression.

Riccio A.A., Sullivan E.D, & Copeland W.C. (2021). Activation of the SARS-CoV-2 NSP14 3′–5′ exoribonuclease by NSP10 and response to antiviral inhibitors. The Journal of Biological Chemistry, 298(1), 101518.

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