pGT4.5A

Deposited by: Alexandra L. Joyner 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. [1] 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. [1] Derived from pGT4.5 by replacing the En-2 polyadenylation signal with an SV40 polyadenylation signal. (personal communication) 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. [2] 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. (ATCC staff) Medium is 1227 LB plus ampicillin. NCBI gi: 403021 Hosts: E.coli, E.coli HB101, vertebrate cells.