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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): 3.9000000953674320 Vector: pBSL15 (plasmid) Construction: pUC18, pK18 Marker(s):ampR,kanR Construct size (kb): 3.900000095367432 Features: marker(s): ampR
marker(s): kanR
replicon: pMB1
MCS: HindIII...EcoRI
MCS: NheI...HindIII
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Applications
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contains easily purifiable cassette(s) for construction aminoglycoside phosphotransferase kanamycin resistance, neomycin resistance vector permitting positive selection for inserts
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Comments
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Restriction digests of the clone give the following sizes (kb): EcoRI--2.8, 1.2; BamHI--2.75, 1.3; HindIII--2.7, 1.4. The nptII gene was modified by site-directed mutagenesis to remove PstI and SphI sites. An inverted repeat of the 271 bp NdeI/EcoRI fragment of pUC18 surrounds the kanR cassette. If this vector is ligated without a stuffer fragment, this inverted repeat prevents replication, thus providing positive selection for plasmids with inserts. Plasmid with an easily purified kanR cassette (flanked by inverted repeats of the pUC18 multiple cloning sequence) for construction. pBSL15 and pBSL86 differ in the orientation of the repeats.
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Media
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LB plus ampicillin (50 mcg/ml) plus kanamycin (50 mcg/ml)
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References
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Alexeyev MF. Three kanamycin resistance gene cassettes with different polylinkers. BioTechniques 18: 52-55, 1995. PubMed: 7702853
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