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| 产品名称： | pBAD18-Kan |
|---|---|
| 商品货号： | TS168904 |
| Designations： | pBAD18-Kan |
| Depositors： | L Guzman, J Beckwith |
| Biosafety Level： | 1 |
| Host： | Escherichia coli DH5α |
| Vector Information： | Intact vector size: 5.437
Type of vector: phagemid Cloning sites: NheI EcoRI SacI KpnI XbaI SalI PstI SphI Polylinker sites: NheI EcoRI SacI KpnI SmaI BamHI XbaI SalI AccI PstI SphI HindIII Construction: pBAD18, kanR (pUC4K) Host range: Escherichia coli Features (with orientation and position when available): regulator: araC, ←, 3346-4224 operator: O2, 4253-4270 promoter: araC, ←, 4375-4403 operator: O1, 4411-4432 other: CAP site, 4454-4467 operator: I2 + I1, 4463-4501 promoter for expression: arabinose BAD, →, 4500-4527 MCS: NheI...HindIII, ->, 4550-4612 transcription terminator: rrnB T1 + T2, →, 4613-5038 marker(s): kanR, →, 123-938 replicon: M13, 1417-1875 replicon: pMB1, 1881-2578 |
| Applications： | Expression vector
Produces protein arabinose regulator
Vector containing primer sites useful for sequencing |
| Comments： | Restriction digests of the clone give the following sizes (kb): EcoRI--5.4; HindIII--3.9, 1.5; PstI--5.4. Cultures should be grown in minimal media for more reproducible induction of expression. Expression is induced in glycerol-containing media by addition of arabinose. Expression is repressed by addition of glucose or other catabolites. One of several tightly controlled expression vectors (ATCC® 87393-87402™) regulated by the arabinose operon. The vectors differ in replicon, antibiotic resistance marker, multiple cloning site and mechanism of initiation of translation. The following primers can be used for sequencing of cloned inserts: 5 primer (27 - 8 bp upstream of the NheI site) 5-CTGTTTCTCCATACCCGTT-3; and one of two 3 primers: 3 primer 1 (2 - 19 bp downstream of the HindIII site) 5-CTCATCCGCCAAAACAG-3; 3 primer 2 (17 - 33 bp downstream of the HindIII site) 5-GGCTGAAAATCTTCTCT-3. Cloned inserts must provide a translation initiation sequence (ATG) and ribosome binding site for expression. Plasmid contains bla (ampR) sequences surrounding the kanR gene which could promoter recombination if this plasmid is used in combination with other compatible ampR plasmids. Recombination can be avoided by the use of recA host strains, or it can be used to advantage to intentionally exchange markers among plasmids. |
| Media： | ATCC® Medium 1236: LB Medium (ATCC medium 1065) with 25 mcg/ml kanamycin |
| Growth Conditions： | Temperature: 37°C |
| References： | Guzman LM, et al. Tight regulation, modulation, and high-level expression by vectors containing the arabinose PBAD promoter. J. Bacteriol. 177: 4121-4130, 1995. PubMed: 7608087 |
| Shipped： | frozen |
| Shipping Information： | Plasmid in Escherichia coli |