pJQ200 (ATCC® 77482)

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

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Designations pJQ200
Depositors MF Hynes
Biosafety Level 1
Host
Distribution host: Escherichia coli DH10B
Vector Information
Size (kb): 4.9000000953674320
DESCRIPTION OF VECTOR:
Intact vector size: 4.900
Type of vector: plasmid
Cloning sites: SmaI
Polylinker sites:
Construction: P15A ori, sacB, RP4, gtmR
Host range: broad host range; Escherichia coli
Features (with orientation and position when available):
replicon: oriV, p15A
other: traJ, <-
marker(s): sacB, ->
replicon: oriT
marker(s): gtmR, <-
Vector: pJQ200 (plasmid)
Construction: P15A ori, sacB, RP4, gtmR
Marker(s):sacB,gtmR
Construct size (kb): 4.900000095367432
Features: marker(s): gtmR
marker(s): sacB
other: traJ
replicon: oriT
replicon: oriV, p15A
Applications
integrating vector
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): SmaI--5.2; EcoRI--5.2; HindIII--5.2.
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.
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 - SmaI - P15A - traJ - oriT - sacB.
Media Medium 1885: LB medium (ATCC medium 1065) with 15 mcg/ml gentamycin
Growth Conditions
Temperature: 37.0°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 frozen
Shipping Information Distributed: freeze-dried
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