产品名称: | lambdaNK1325 |
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商品货号: | TS137399 |
Designations: | lambdaNK1325 |
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): 6.0 Gene product: transposase, Tn10, ats1 ats2 (ATS) URA3 Target Gene: transposase, Tn10, ats1 ats2 (ATS) |
Insert Size (kb): | 6.0 |
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-->23.0, 11.5, 6.8, 3.3; HindIII-->23.0, 6.8, 4.0, 3.5, 1.3, 1.2, 0.9; BglII--33.5, 8.2, 2.7; XbaI--26.0, 18.0. Construct useful for insertional mutagenesis in E. coli and Saccharomyces cerevisiae. 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 3.4 kb mini-Tn10 cassette conferring kanamycin resistance and encoding URA3, 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 URA kan - 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 Riles L, et al. Physical maps of the six smallest chromosomes of Saccharomyces cerevisiae at a resolution of 2.6-kilobase pairs. Genetics 134: 81-150, 1993. PubMed: 8514151 |