Restriction digests of the clone give the following sizes (kb): HindIII--3.4, 1.5; NcoI--3.5, 1.4; PvuII--4.9; SalI--2.4, 2.5; BamHI--3.9, 1.0. The insert contains the following restriction sites (approximate kb from the 5' end): HindIII--0.58; NcoI--0.78; PvuII--0.52. Contains the complete coding sequence (nt 40-825 of the sequence record). COS-7 cells transfected with this construct demonstrate good surface expression of the protein. The vector is modified from pcD, with a promoter constructed from the SV40 early promoter and the R segment and U5 sequence of the long terminal repeat of human T-cell leukemia virus type 1 (HTLV-1 LTR). Subcloned from pBS-hCD40L-6A9 (ATCC 79812/3). The 5' BamHI site is derived from the vector and the 3' end is the naturally occurring BamHI site (nt 929 of the published sequence). |