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GeneCorner plasmid details

Last data update: 01 December 2020 04:11 CET

Plasmid name: pCB1003 (LMBP 8050)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p8050.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: -
Promoter: Aspergillus nidulans tryptophan C promoter (trpC)
Escherichia coli lac operon promoter
Ribosome
binding site:
Ribosome binding site (RBS) of the Escherichia coli lac Z gene (lacZ)
Terminator: -
Selection marker: Ampicillin (amp)
Hygromycin (hyg)
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli
Magnaporthe sp.
Ascomycete fungi
Parental clone: pCSN43; pUC19
Further information: The plasmid was constructed as follows: 1) the coding sequence of the hygromycin resistance gene from pCSN43 was mutated through three rounds of megaprimer mutagenesis, eliminating the EcoRI (GAgTTC), PstI (CTGgAG), SstII (CCGCGc) and BamHI (GGtTCC) restriction sites and eliminating the ClaI site near the start codon. The latter mutation changed the Kozak sequence from ATCGATATG to ATCcAaATG. 2) the mutated hygromycin cds from pCSN43 was PCR amplified, adding EcoRI, SalI and HpaI restriction sites at both ends. 3) the amplified hygromycin cds was ligated as an EcoRI fragment into the EcoRI site of pUC19.
EMBL Accession number: -
Latest sequence update: 14/01/2013
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BamHI, EcoRI, NdeI and PvuI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Prof. Dr K. McCluskey(1).
(1) FGSC, University of Missouri, Kansas City, USA
Plasmid reference: Caroll et al., Fungal Genet. Newsl. 41 (1994), 22
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 DH5α
Host reference: Focus 8 (1986), 9
Related host reference: Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051]
Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187]
Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791]
Rodriguez-Quinones et al., Focus 15 (1993), 110-112
Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660]
Cultivation medium: LB-Lennox + ampicillin (100 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
Cultivation remark: -
Other culture collection numbers: -

Refer in your Materials and Methods:

pCB1003 (LMBP 8050) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr K. McCluskey and was published in Caroll et al., 1994.

Note: Up-to-date, validated data are enclosed with the biological material. Nevertheless, these data are a snapshot at a given moment; further updates are always possible.

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