pJQ210SK (ATCC® 77489)

Applications: integrating vectorvector for constructing genomic librariesvector permitting construction of fusion proteinsvector permitting positive selection for integrationvector permitting visual detection of recombinants  /  Depositors: MF Hynes

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Designations pJQ210SK
Depositors MF Hynes
Biosafety Level 1
Vector Information
Size (kb): 7.1999998092651370
Vector: pJQ210SK (cosmid)
Promoters: Promoter lac
Construction: P15A ori, sacB, RP4, gtmR, cos, pBluescriptSK MCS
Marker(s):sacB,gtmR
Construct size (kb): 7.199999809265137
Features: insert detection: lac
marker(s): gtmR, sacB
promoter: lac
replicon: P15A, oriT
Applications
integrating vector
vector for constructing genomic libraries
vector permitting construction of fusion proteins
vector permitting positive selection for integration
vector permitting visual detection of recombinants
Comments
Restriction digests of the clone give the following sizes (kb): SstI--7.9; KpnI--6.0, 1.9; BamHI--7.9.
Replicates only in enterobacteria, and so functions as a suicide vector in other Gram negative hosts.
A single BglII site within the gentamicin resistance gene (gtmR) allows insertion of other resistance genes (e.g. as a BamHI cassette).
When induced by media containing 5% sucrose, sacB is lethal in a wide range of Gram negative bacteria, and thus permits selection for loss of the vector.
IMPORTANT: To prevent amplification of a rearranged and/or deleted cosmid, we recommend streaking on LB + amp plates at 30C and picking small colonies for liquid culture.
Suicide vector permitting gene replacement and mobilization into a wide range of Gram negative bacteria. Allows positive selection for integration.
The order of the major features of the plasmid is: gtmR - SstI/MCS/KpnI - P15A - cos - traJ - oriT - sacB.
Media ATCC® Medium 1885: LB medium (ATCC medium 1065) with 15 mcg/ml gentamycin
Growth Conditions
Temperature: 30.0°C
References

Quandt J, Hynes MF. Versatile suicide vectors which allow direct selection for gene replacement in gram-negative bacteria. Gene 127: 15-21, 1993. PubMed: 8486283

Shipped freeze-dried
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