Last data update: 24 January 2024 16:39 CET
Plasmid name: pEF1hBCL2 (LMBP 5105)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p5105.gb
(View with Genome Compiler) p5105.txt p5105.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human B-cell leukemia/lymphoma 2α cDNA (BCL2, Bcl-2) V5 epitope; C-terminal Histidine tag (His-tag); C-terminal |
| Promoter: | Human elongation factor 1α promoter (EF1α) Phage T7 gene 10 promoter (T7g10) Simian virus 40 early promoter (SV40 early) Escherichia coli lac operon promoter |
| Ribosome binding site: |
- |
| Terminator: | Bovine growth hormone polyadenylation signal (BGH polyA) Simian virus 40 polyadenylation signal (SV40 polyA) |
| Selection marker: | Ampicillin (amp) Neomycin (neo; G418) |
| Replicon: | Escherichia coli plasmid pMB1 origin Phage f1 origin Simian virus 40 bidirectional origin (SV40) |
| Host range: | Escherichia coli Mammalian cells; SV40 permissive cells |
| Parental clone: | pEF1/V5-HisA; pCRhBCL2 |
| Further information: | The plasmid was constructed by inserting the EcoRI fragment of pCRhBCL2, containing the human BCL2 cDNA, inclusive its 5' and 3' UTR, in the EcoRI opened pEF1/V5-HisA vector. As a result, the C-terminal V5 epitope and histidine tag are not expressed. The plasmid is designed for high-level expression of human BCL2 under control of the human EF1α promoter in mammalian cells. The plasmid carries the ampicillin resistance gene for selection in E. coli and the neomycin resistance gene for selection in mammalian cells. The presence of a second enhancer in the SV40 early promoter was shown by sequencing at LMBP. Unlike pcDNA3, pEF1hBCL2 has not been shown to provide resistance to kanamycin. The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727086.1. The human BCL2 EcoRI fragment was sequenced by BCCM/GeneCorner and found identical to the DNA sequence described in the EMBL Nucleotide Sequence Database accession number BC027258.1, except for position 189 of the coding sequence, resulting in a silent mutation. Other name of the plasmid is pEF1/V5-HisABcl2. |
| EMBL Accession number: | BC027258.1, view at EMBL, GenBank, DDBJ LT727086.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 10/11/2006 |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BamHI, EcoRI, MlsI and PvuII. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Prof. Dr P. Vandenabeele(1) (2). It was constructed by E. Van Damme(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 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pEF1hBCL2 (LMBP 5105) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr P. Vandenabeele . |
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.