pNK1207 (ATCC® 77354)

Organism: Escherichia coli (Migula) Castellani and Chalmers  /  Clone Type: Clone  /  Depositors: N Kleckner

Permits and Restrictions

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Designations pNK1207
Species Escherichia coli (Migula) Castellani and Chalmers
Depositors N Kleckner
Vector
Construct size (kb): 10.10000038146973
Insert
DNA: genomic
Insert lengths(kb): 8.0
Gene product: ATP phosphoribosyltransferase [hisD]
Insert Size (kb) 8.0
Media Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Biosafety Level 1
Comments
Restriction digests of the clone give the following sizes (kb): EcoRI--4.2 (doublet), 2.1; HindIII--10.5; PstI--10.5.
Construct permitting translational fusions of lacZ to target genes. The transposase must be supplied in trans (pNK2882, ATCC 77226).
LacZ is expressed after transpositions in the correct orientation and reading frame into an expressed target gene.
Derived from pNK81 (ATCC 77337) by replacing 1.050 kb of uncharacterized DNA and the his promoter with 4 tandem repeats of the rrnB transcriptional terminator and replacing Tn10 with the mini-Tn1-LK.
Contains the mini-Tn10-LK cassette that consists of the following features: IS10R - nt 63 - promoterless lacZ element starting at codon 8 - nt 3140 - kanR - IS10L - nt 4900.
The order of the major features in this plasmid is: EcoRI - rrnB terminators - Tn10-LK in hisG - hisD - EcoRI - pMB1 ori - ampR.
Classification Enterobacteriaceae, Escherichia
References

Kleckner N, et al. Uses of transposons with emphasis on Tn10. Methods Enzymol. 204: 139-180, 1991. PubMed: 1658561

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