Size (kb): 9.5000000000000000
Vector: pGT4.5A (plasmid)
Construction: En-2, lacZ, neo, pUC18
Construct size (kb): 9.5
Features: insert detection: lacZ'
terminator: SV40 polyadenylation
contains sequence engrailed-2
vector permitting construction of fusion proteins
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.
Skarnes WC, et al. A gene trap approach in mouse embryonic stem cells: the lacZ reporter is activated by splicing, reflects endogenous gene expression, and is mutagenic in mice. Genes Dev. 6: 903-918, 1992. PubMed: 1592261
Gossler A, et al. Mouse embryonic stem cells and reporter constructs to detect developmentally regulated genes. Science 244: 463-465, 1989. PubMed: 2497519
Alexandra L Joyner, personal communication