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| 产品名称： | pHU10 HIS3 --> URA3 converter |
|---|---|
| 商品货号： | TS155795 |
| Designations： | pHU10 HIS3 --> URA3 converter |
| Depositors： | FR Cross |
| Biosafety Level： | 1 |
| Host： | Escherichia coli HB101 |
| Vector Information： | Construct size (kb): 6.60
Type of vector: Phagemid
Host range: Saccharomyces cerevisiae, Escherichia coli Features (with orientation and position when available): replicon: f1, →
marker(s): cmlR, ←
replicon: pMB1
restriction site: XhoI, SalI |
| Applications： | YI-type (integrating) shuttle vector host modification marker swap vector HIS3 --> URA3 |
| Comments： | Restriction digests of the clone give the following sizes (kb): HindIII--6.5; HindIII/PstI--5.0, 1.1, 0.5. To convert the host phenotype from HIS3 to URA3, transform with the SmaI+XhoI digested vector and select for Ura+ transformants. Some combinations of marker swap plasmids and target locus may result in relatively high reversion rates. In most but not all cases the frequencies of successful convertants are greater than 30%. A marker swap vector designed to change the S. cerevisiae host phenotype by one-step gene disruption of the HIS3 gene with the URA3 and kanR markers. When swapping markers on an episomal plasmid, appropriate phenotype may result from loss of the plasmid unless a second selectable or scorable marker is used to ensure plasmid maintenance. Vector was constructed by replacing an internal HindIII fragment of HIS3 with a SmaI fragment containing the URA3 and kanR coding sequences. URA3 and HIS3 are in the opposite orientation. |
| Media： | ATCC Medium 1065 (see below) plus kanamycin (50.0 mcg/ml) plus chloramphenicol (25.0 mcg/ml) ATCC Medium 1065: Tryptone (Difco 0123), 10.0 g Yeast Extract (Difco 0127), 5.0 g NaCl, 10.0 g Distilled water, 1.0 L
|
| Growth Conditions： | Temperature: 37°C |
| References： | Cross FR. Marker swap plasmids: convenient tools for budding yeast molecular genetics. Yeast 13: 647-653, 1997. PubMed: 9200814 |
| Related Products： | Component of: ATCC 87561 |
| Shipped： | freeze-dried |