pDOI-5 (ATCC® 87058)

Applications: Expression vector for creating transgenic mice Shuttle vector  /  Depositors: C Benoist

Designations pDOI-5
Permits and Restrictions

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Depositors C Benoist
Biosafety Level 1
Host Escherichia coli MC1061; Roche 3943B
Vector Information
Intact vector size: 7.240
Type of vector: plasmid
Cloning sites: BamHI EcoRI
Construction: pKCR
Host range: Escherichia coli; mouse

Features (with orientation and position when available):
promoter: Ea, →, 5046-7240
restriction site: BamHI, 1
other: beta-globin 3' sequence, →, 1-1200
restriction site: EcoRI, 638
replicon: pMB1, ←, 3226
marker(s): ampR, ←, 3984-4847
Vector: pDOI-5 (plasmid)
Promoters: Promoter Ea
Expression vector for creating transgenic mice
Shuttle vector
Restriction digests of the clone give the following sizes (kb): BamHI--7.0; EcoRI--7.0; XbaI--7.0.

The promoter is followed by a portion of the rabbit beta-globin gene, which provides a splice and a polyadenylation signal and is thought to provide a nuclear export signal.

The BamHI cloning site may be prone to problems with cryptic splice donors leading to truncated transcripts.
Elimination of non-essential plasmid sequence before microinjection of a recombinant vector may increase expression efficiency
in the mouse.

After insertion of the target gene, the pBR322 portion of the vector can be digested with one of the following enzymes to provide a linear molecule for microinjection: AatII, BglI, XbaI, NruI or HhaI.

Shuttle expression vector used to target expression of a cloned gene to murine cells which normally display MHC class II molecules.

The order of the major features in the plasmid is: Ea promoter - BamHI - beta-globin sequence/EcoRI - XhoI - pMB1 ori - ampR - XbaI.
Media ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37°C

Kouskoff V, et al. A vector driving the expression of foreign cDNAs in the MHC class II-positive cells of transgenic mice. J. Immunol. Methods 166: 287-291, 1993. PubMed: 7507147

Shipped frozen