Restriction digests of the clone give the following sizes (kb): BamHI--3.8, 3.3, 1.8, 0.45, 0.2; HindIII--9.5; XbaI--4.6, 3.1, 1.0, 0.8. Gene trap vector designed to identify and mutate endogenous mammalian genes by integration and production of a fusion protein with the lacZ reporter gene. Can also be used to monitor endogenous gene expression. Identified cDNA sequences spanning the lacZ splice junction can be cloned using the published sequence of the En-2 splice acceptor and the rapid amplification of cDNA ends (RACE) protocol. Derived from pGT4.5 by replacing the En-2 polyadenylation signal with an SV40 polyadenylation signal. The order of the major features in this plasmid is: pUC18 - HindIII - En-2 intron - splice acceptor - En-2 homeobox exon - lacZ - SV40 polyadenylation - human beta-actin promoter - neoR - SV40 polyadenylation. |
