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| 产品名称： | pAG408 |
|---|---|
| 商品货号： | TS171375 |
| Designations： | pAG408 |
| Depositors： | C Guzman |
| Biosafety Level： | 1 |
| Host： | Escherichia coli CC118 lambda pir+ |
| Vector Information： | Size (kb): 5.7
Type of vector: plasmid Cloning sites: Polylinker sites: BstXI NotI SpeI SmaI XmaI ClaI ApaI KpnI Construction: pASV1, pBluescript KS+, pBSL202 Host range: Escherichia coli Vector: pAG408 (plasmid)
Marker(s): ampR,kanR,gtmR
Features (with orientation and position when available): coding sequence: Tn5 transposase, → marker(s): ampR, → replicon: oriT RP4, transfer origin replicon: R6K ori other: atpE reporter group: modified gfp gene, → coding sequence: aphA-3, → marker(s): gtmR, → |
| Applications： | Integrating vector
Suicide vector
Vector permitting positive selection for integration |
| Comments： | The vector is a suicide mini-transposon delivery plasmid and R6K based, being unable to survive in recipients lacking the Pir protein. Selection for the kanamycin or gentamycin resistance encoded by the mini-transposons yields recipients in which the mini-transposon, but not the cognate transposase gene present on the suicide plasmid, has integrated into one of the endogenous replicons. The modified GFP exhibits a 45-fold-increased fluorescent signal compared with that of the natural GFP. The enhanced sensitivity of this vector, gfp-based promoter-probe mini-transposon, allows the direct detection of transconjugants harbouring monocopy transcriptional fusions by means of a UV handy-lamp at 366 nm by the naked eye and the detection of single gfp-tagged bacterial cells by epifluorescence microscopy. Restriction digests of the clone give the following sizes (kb): NotI--5.7; ClaI--5.7; KpnI--4.1, 1.6. |
| Media： | ATCC® Medium 1948: LB medium (ATCC medium 1065) with 50 mcg/ml ampicillin and 20 mcg/ml kanamycin |
| Growth Conditions： | Temperature: 37°C |
| References： | Suarez A, et al. Green fluorescent protein-based reporter systems for genetic analysis of bacteria including monocopy applications. Gene 196: 69-74, 1997. PubMed: 9322742 |
| Shipped： | frozen |