Promoter: The promoter driving your CAR expression cassette is placed here.
Kozak: Kozak consensus sequence. It is placed in front of the start codon of the ORF of interest because it is believed to facilitate translation initiation in eukaryotes.
CD8-leader: Leader signal peptide of T-cell surface glycoprotein CD8 alpha chain. Directs transport and localization of the protein to the T-cell surface.
scFv: Single chain variable fragment derived from a monoclonal antibody of known specificity. Recognizes cells in an antigen-specific manner.
Hinge: Extracellular hinge region of the CAR. Connects scFv with the transmembrane region providing stability and flexibilty for efficient CAR expression and function; enhances efficiency of tumor recognition; improves expansion of CAR-T cells.
Transmembrane domain: Transmembrane domain of the CAR. Anchors the CAR to the plasma membrane and bridges the extracellular hinge as well as antigen recognition domains with the intracellular signaling region; enhances receptor expression and stability.
Costimulatory domain: Intracellular costimulatory domain of the CAR. Improves overall survival, proliferation, and persistence of activated CAR-T cells.
CD3zeta: Intracellular domain of the T cell receptor-CD3ζ chain. Acts as a stimulatory molecule for activating T cell-mediated immune response.
SV40 late pA: Simian virus 40 late polyadenylation signal. It facilitates transcriptional termination of the upstream CAR expression cassette.
CMV promoter: Human cytomegalovirus immediate early promoter. It drives the ubiquitous expression of the downstream marker gene.
Marker: A drug selection gene (such as neomycin resistance), a visually detectable gene (such as EGFP), or a dual-reporter gene (such as EGFP/Neo). This allows cells transduced with the vector to be selected and/or visualized.
BGH pA: Bovine growth hormone polyadenylation. It facilitates transcriptional termination of the upstream ORF.
Ampicillin: Ampicillin resistance gene. It allows the plasmid to be maintained by ampicillin selection in E. coli.
pUC ori: pUC origin of replication. Plasmids carrying this origin exist in high copy numbers in E. coli.