Last data update: 24 January 2024 16:39 CET
Plasmid name: pSP64ble1 (LMBP 2421)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p2421.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: | pSP64; pUT332 |
| Further information: | The plasmid was constructed as follows: 1) pUT332 was digested with KpnI; the 3' sticky ends were blunted with T4 DNA polymerase and synthetic HindIII linkers (5'-GGAAGCTTCC-3') were added; 2) This intermediate construction was then cut with HindIII and the fragment containing the phleomycin resistance expression cassette was cloned into the unique HindIII site of pSP64. 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 pSp64Phleo1. |
| EMBL Accession number: | - |
| Latest sequence update: | 21/12/1994 |
| 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: |
pSP64ble1 (LMBP 2421) 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.