pcaggs vector - Anopheles Database

The vector for the receptor binding domain (RBD) of the spike (S) glycoprotein gene from severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2), Wuhan-Hu-1 (GenBank: MN908947) was designed by fusing the N-terminal S protein signal sequence to the spike RBD (amino acids 319 to 541) with a C-terminal hexa-histidine tag.

The sequence was codon optimized for mammalian expression and subcloned into the pCAGGS mammalian expression vector under the AG promoter. NR-52309 contains the beta-lactamase gene, bla, to provide transformant selection through ampicillin resistance in Escherichia coli (E. coli). The resulting size of the plasmid is approximately 5490 base pairs. The complete plasmid sequence is provided on the BEI Resources webpage. The plasmid was produced in Escherichia coli and extracted.

Note: For a detailed protocol and list of related items, see https://labs.icahn.mssm.edu/krammerlab/covid-19

The S glycoprotein mediates viral binding to the host angiotensin converting enzyme 2 (ACE2). This protein forms a trimer, and when bound to a host receptor allows fusion of the viral and cellular membranes. The S protein is a target for neutralizing antibodies.

Each vial contains plasmid DNA in TE buffer (10 mM Tris-HCl, 1 mM EDTA, pH 8.0). The vial should be centrifuged prior to opening. Note: The contents of the vial should be used to replicate the plasmid in E. coli prior to mammalian expression.