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Permits
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These permits may be required for shipping this product:
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Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
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Vector Information
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Size (kb): 5.8460001945495610 Vector: pEX100T (plasmid) Construction: pUC19Sce, pEM5, pMOB2 Marker(s):ampR,sacB Construct size (kb): 5.846000194549561 Features: marker(s): ampR
marker(s): sacB
replicon: oriT
replicon: pMB1
restriction site: I-SceI
restriction site: SmaI
transcription terminator: rrnB T1
transcription terminator: rrnB T2
coding sequence: lacZalpha
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Applications
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vector permitting positive selection for integration levansucrase vector permitting visual detection of recombinants
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Comments
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Restriction digests of the clone give the following sizes (kb): ClaI--5.6, 0.40; ScaI--6.0; SmaI--6.0; AvaI--6.0. The ClaI restriction recognition site located at bp 2093 is subject to dam methylation (gATCGAT) in its distribution host DH5alphaF'. The ampR marker can easily be replaced by other antibiotic resistance-encoding cassettes by cloning into the unique ScaI site contained within the bla gene. E. coli strains containing sacB are sucrose sensitive on LB medium + 5% sucrose. It is important to monitor sucrose sensitivity at each cloning step, as spontaneous mutations can accumulate during the process which then confer sucrose resistance. Vector to construct mutants in Pseudomonas aeruginosa.
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References
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Schweizer HP, Hoang TT. An improved system for gene replacement and xylE fusion analysis in Pseudomonas aeruginosa. Gene 158: 15-22, 1995. PubMed: 7789804
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Shipping Information
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Frozen glycerol stock of E. coli containing the plasmid
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