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| 产品名称： | pBAD18sc |
|---|---|
| 商品货号： | TS169168 |
| Designations： | pBAD18sc |
| Depositors： | L Guzman, J Beckwith |
| Biosafety Level： | 1 |
| Vector Information： | Size (kb): 5.5630002021789550 Vector: pBAD18sc (phagemid) Promoters: Promoter araC Construction: pMJC105, pDHB60 Marker(s):ampR,araC Construct size (kb): 5.563000202178955 Features: marker(s): ampR operator: I2 + I1 operator: O1 operator: O2 other: CAP site promoter: araC promoter for expression: arabinose BAD regulator: araC replicon: M13 replicon: pMB1 MCS: EcoRI...HindIII ribosome-binding site: Shine-Dalgarno sequence transcription terminator: rrnB T1 + T2 coding sequence: 5 portion of araB |
| 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--5.6; HindIII--5.6; PstI--4.6, 1.1. 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. Vector contains the Shine-Dalgarno box and 5 amino acids of the araB gene, but does not necessarily form fusion proteins with cloned inserts. Higher expression may still be achieved due to the second SD box despite out-of-frame cloning. Cloned inserts should provide a translation initiation sequence (ATG) and ribosome binding site for expression. |
| Media： | ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin |
| Growth Conditions： | Temperature: 37.0°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： | freeze-dried |