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| 产品名称： | pOF216 |
|---|---|
| 商品货号： | TS165366 |
| Designations： | pOF216 |
| Depositors： | F Hosoda, M Ohki |
| Biosafety Level： | 1 |
| Vector Information： | Size (kb): 12.5000000000000000 Vector: pOF216 (plasmid) Promoters: Promoter lac Construction: pKP1013, pUC13 Marker(s):spcR Construct size (kb): 12.5 Features: insert detection: lacZ marker(s): spcR promoter: lac replicon: oriF |
| Applications： | vector with low copy number |
| Comments： | Restriction digests of the clone give the following sizes (kb): BamHI--12.5; HindIII--12.5. The EcoRI site is available for cloning but the lacPO N-terminal region will be deleted. The polylinker is inserted in frame within the lacZ gene facilitating selection for blue and white colonies. Ligating lambda cosR adapters onto the termini of potential inserts and cosL adapters onto the linearized vector enhances the step of circularization of large molecules. This is a low copy number mini-F based plasmid vector with rare restriction sites in its polylinker for cloning inserts of up to 100 kb. The depositor recommends adding spectinomycin to the media at a concentration of 20-30 ug/ml at the first establishment of spcR transformants and 15 ug/ml thereafter. |
| Media： | ATCC Medium 1065 (see below) plus 15.0 mcg/ml tetracycline
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: 37.0°C |
| References： | Hosoda F, et al. An F factor based cloning system for large DNA fragments. Nucleic Acids Res. 18: 3863-3869, 1990. PubMed: 2197597 Mori H, et al. Structure and function of the F plasmid genes essential for partitioning. J. Mol. Biol. 192: 1-15, 1986. PubMed: 3029390 Maki S, et al. An amber replication mutant of F plasmid mapped in the minimal replication region. Mol. Gen. Genet. 191: 231-237, 1983. PubMed: 6353162 Fumie Hosoda, personal communication |
| Shipped： | freeze-dried |