Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Also contains pAdD26SV(A)-3 which was cotransformed to express murine Dhfr. Both plasmids were digested with EcoRI before transformation.
This clone was selected after growth in 250 nM methotrexate.
The order of the major features in this plasmid is: SalI - SV40 promoter - HindIII - IFNG - SV40t - SV40 polyadenylation - EcoRI - ampR - ori.
Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
Fiers WC, Allet B. DNA sequences, recombinant DNA molecules and processes for producing human gamma interferon-like polypeptides in high yields. US Patent 5,004,689 dated Apr 2 1991
