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lambdaNK1327

货号 TS147611
中文名称 null
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产品名称: lambdaNK1327
商品货号: TS147611
Designations: lambdaNK1327
Species: Escherichia coli (Migula) Castellani and Chalmers
Depositors: N Kleckner, S Gottesman
Applications:
produces protein transposase, Tn10, ats1 ats2 (ATS)
Vector:
Construct size (kb): 0.0
Insert:
DNA: genomic
Insert lengths(kb): 4.5
Gene product: transposase, Tn10, ats1 ats2 (ATS)
Target Gene: transposase, Tn10, ats1 ats2 (ATS)
Insert Size (kb): 4.5
Biosafety Level: 1

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Comments:
Restriction digests of the clone give the following sizes (kb): EcoRI--26.0, 9.8, 5.0, 3.1; HindIII-->23.0, 6.2, 3.8, 3.3, 1.2, 0.95, 0.6; BglII--33.5, 7.8, 2.35, 0.7; XbaI--26.0, 18.0.
Construct useful for general transposon mutagenesis in Escherichia coli. It also permits generation of promoter fusions of target genes to Plac-UV5, if the transposon inserts in the correct orientation.
The vector has mutations in replication genes, the repressor, and the integration system. Genotype: b522 cI857 Pam80 nin5. The EcoRI cloning site is between bp 21226 and 26104.
Contains the ats1 ats2 transposase gene that permits relaxed insertion specificity (altered target specificity, ATS). Expression is regulated by the Ptac promoter, inducible by IPTG.
The transposase segment extends from IS10R to the EcoRI site at nt 3140 of Tn10, with a deletion of nucleotides 1329-1942 to remove the transposase binding site.
Contains a 1.9 kb mini-Tn10 cassette conferring kanamycin resistance, bounded by inverted repeats of the outermost 70 bp of IS10R and embedded in 40 bp of lambdacI terminating in HindIII sites.
The order of the major features in this bacteriophage is: A-J - EcoRI - mini-Tn10 kan Plac-UV5 - ATS transposase - Ptac - EcoRI - b522 - cI857 - P80 - nin5.
Classification: Enterobacteriaceae, Escherichia
References:

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