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. 2021 Jan 20;11(2):e3883. doi: 10.21769/BioProtoc.3883

Figure 10. MHC Vector Construction.

Figure 10.

A. Plasmid map of non-replicating lentiviral backbone vector pUS. A ubiquitous chromatin opening element (UCOE) precedes the spleen focus forming virus (SFFV) promoter (Adamson et al., 2016). An MluI-SbfI cloning site is located after the SFFV promoter. B. Plasmid map of an example MHC I vector, A2_uSFFV. A2_uSFFV vector was constructed by digesting the pUS backbone vector with MluI and SbfI and inserting the HLA-A*02:01 gene-P2A-human β2 microglobulin gene cassette by Gibson assembly. C. Schematic diagram of an MHC I gene-P2A-human β2 microglobulin gene cassette. D. Schematic diagram of an MHC II α gene-P2A-MHC II β gene cassette. The blue brackets indicate the positions of the 5’ and 3’ DNA fragment ends added to MHC gene sequences..