Constructing sequences for expression plasmids

Data files contain construct sequences for expression plasmids related to the manuscript by Heider et al .The pRRL.sin.EF1α.eGFP plasmid was derived from an HIV-1 based pRRL.sin backbone plasmid and the enhanced green fluorescent (eGFP) protein was placed under the control of an elongation factor 1 α promoter (EF1α) (44). Then, pRRL.sin.EF1α.PPI.P2A.eGFP was generated from pRRL.sin.EF1α.eGFP plasmid. The encoding sequence for human preproinsulin (hPPI, aa1-110) was inserted upstream of the eGFP sequence and separated from it by a peptide 2A cleavage site sequence. pRLL.sin.HLA-DRα.eGFP.EFS.mTagBFP2 plasmid was then generated from pRRL.sin.EF1α.PPI.P2A.eGFP, by replacing the EF1α promoter with that of a HLA-DRα. The mTagBFP2 sequence (45) from pRRL.sin.EF1α.Hygro.P2A.mTagBFP2 (46) plasmid (AddGene #99374) was placed under the control of an intron-less EF1α promoter (EFS) (47) and then inserted. The hPPI sequence was then added upstream of the eGFP sequence and separated by a P2A cleavage site sequence to generate pRLL.sin.HLA-DRα.PPI.P2A.eGFP.EFS.mTagBFP2. pRRL.sin.HLA-DRα.pp65.P2A.EFS.mTagBFP2 was derived from the pRLL.sin.HLA-DRα.PPI.P2A.eGFP.EFS.mTagBFP2 plasmid, but the sequence encoding NY-ESO-1157-165 CMV pp65495-503 with linkers (48, 49) replaced the hPPI2-24 sequence so that a NY-ESO-1157-165 pp65495-503 PPI24- 110 fusion protein was encoded.