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GeneCorner plasmid details

Last data update: 24 January 2024 16:39 CET

Plasmid name: pSP64ble3 (LMBP 2420)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p2420.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Transposon Tn5 bleomycin gene (ble) conferring resistance to antibiotics of the bleomycin family: bleomycin, phleomycin, zeomycin
Promoter: Escherichia coli lac operon promoter
Phage SP6 promoter
Saccharomyces cerevisiae translation elongation factor 1α promoter (TEF1α)
Ribosome
binding site:
-
Terminator: Saccharomyces cerevisiae iso-1-cytochrome C terminator (CYC1)
Selection marker: Ampicillin (amp)
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli
Parental clone: pSP64ble1
Further information: This plasmid was derived from pSP64ble1 by two consecutive partial BamHI digestions at nucleotide positions 14 and 912; the 5' sticky ends were filled in with Klenow DNA polymerase and ligated again.
The plasmid carries the phleomycin resistance expression cassette, which is removable as a HindIII fragment without internal BamHI sites.
When cloning a fragment downstream from the lac promoter it may be advisable to use lacI(q) strains in order to prevent fortuitous expression of a possibly noxious polypeptide.
Other name of the plasmid is pSp64-Phleo3.
EMBL Accession number: -
Latest sequence update: 22/12/1994
Authenticity test: The plasmid still needs to be subjected to the authenticity test.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Dr J. De Molder(1) (2) and Prof. Dr R. Contreras(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: -
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: Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660]
Rodriguez-Quinones et al., Focus 15 (1993), 110-112
Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791] [DOI: 10.1016/s0022-2836(83)80284-8]
Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187]
Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051]
Cultivation medium: LB-Lennox + ampicillin (100 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
Other culture collection numbers: -

Refer in your Materials and Methods:

pSP64ble3 (LMBP 2420) is available at BCCM/GeneCorner. This plasmid was deposited by Dr J. De Molder and Prof. Dr R. Contreras .

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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