pGT4.5A (ATCC® 77329)

Applications: contains sequence engrailed-2reporter constructshuttle vectorvector permitting construction of fusion proteins  /  Depositors: AL Joyner

Permits and Restrictions

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Designations pGT4.5A
Depositors AL Joyner
Biosafety Level 1
Vector Information
Size (kb): 9.5000000000000000
Vector: pGT4.5A (plasmid)
Construction: En-2, lacZ, neo, pUC18
Marker(s):ampR
Construct size (kb): 9.5
Features: insert detection: lacZ'
marker(s): ampR
replicon: pMB1
terminator: SV40 polyadenylation
Applications
contains sequence engrailed-2
reporter construct
shuttle vector
vector permitting construction of fusion proteins
Comments
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.
Media ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C
References

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

Shipped freeze-dried
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