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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): 7.3579998016357420 Vector: pJIR418 (plasmid) Promoters: Promoter lac Construction: pUC18, pIP404 replicon, pJIR229 ermBP, pJIR62 catP Marker(s):cmlR,eryR Construct size (kb): 7.357999801635742 Features: insert detection: lacZ'
marker(s): cmlR, eryR
promoter: lac
replicon: pMB1, pIP404
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Applications
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expression vector shuttle vector vector permitting construction of fusion proteins 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): EcoRI--7.4; HindIII--7.4; SmaI--5.0, 2.4. The vector contains the following restriction sites (position in bp): EcoRI--1; HindIII--57; PvuI--175; FspI--195; NarI--216; Tth111I--397; SpeI--3235; NheI--3549; NaeI--3653; HpaI--4206; EcoRV--4723; SmaI--4984; AvaII--5059; ScaI--5850. The genes for chloramphenicol and erythromycin resistance (catP and ermBP, respectively) were cloned from C. perfringens and can be used for selection in both C. perfringens and E. coli. EcoRV and ScaI restriction sites can be used for cloning and insertional inactivation of catP and ermBP, respectively. The standard reverse sequencing primer is not useful in this vector due to the duplication of the 181 bp region between pUC18 sites PvuII and EcoRI. Shuttle expression vector permitting visual detection of recombinants in E. coli using lacZalpha complementation. Does not contain an intact beta-lactamase (bla) gene. The order of the major features in this plasmid is: 5' lacZ' - EcoRI/MCS/HindIII - 3' lacZ' - pIP404 ori - pIP404 rep - HpaI - catP - SmaI - AvaII - ermBP - pMB1 ori. Location of the major features in this plasmid: lacZ' (nt 7345-310), MCS (nt 1-57), pIP404 ori (approx. nt 1190-1419), pIP404 rep gene (nt 2702-1482), catP (nt 4830-4207), ermBP (nt 5902-5165), pMB1 ori (approx. nt 6780-7000).
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Media
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ATCC Medium 1065 (see below) plus 30.0 mcg/ml chloramphenicol 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
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References
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Sloan J, et al. Construction of a sequenced Clostridium perfringens-Escherichia coli shuttle plasmid. Plasmid 27: 207-219, 1992. PubMed: 1513878
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Shipping Information
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Freeze dried Escherichia coli containing the plasmid.
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