pJQ200SK plasmid in E. coli (ATCC® 77483)

Applications: Integrating vector Vector permitting construction of fusion proteins Vector permitting positive selection for integration Vector permitting visual detection of recombinants  /  Depositors: MF Hynes

Designations pJQ200SK plasmid in E. coli
Permits and Restrictions

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Depositors MF Hynes
Disclosure REPOS
Biosafety Level 1
Host Escherichia coli DH10B
Vector Information Total size of construct (kb): 5.4
Marker: gtmR, sacB
Vector type: plasmid
Cloning sites: SstI BstXI NotI XbaI SpeI BamHI SmaI PstI SalI XhoI ApaI
Construction: pBluescriptSK MCS, P15A ori, sacB, RP4, gtmR
Host range: broad host range; Escherichia coli

Features:
Promoter: lac
Replicon: P15A, oriT 
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): BamHI--5.4; KpnI--3.6, 1.8; SstI--5.4.
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 - SstI/MCS/KpnI - P15A - traJ - oriT - sacB.
Growth Conditions
Temperature: 37°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 In glycerol stock of E. coli containing the plasmid