Last data update: 24 January 2024 16:39 CET
Plasmid name: pEF6mBclxLMycHis (LMBP 5078)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p5078.gb
(View with Genome Compiler) p5078.txt p5078.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Mouse B-cell leukemia/lymphoma 2-like 1 cDNA (BclxL, Bcl2l1); long form Myc 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) Synthetic prokaryotic EM7 promoter 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) Blasticidin (bsd) |
| Replicon: | Escherichia coli plasmid pMB1 origin Phage f1 origin Simian virus 40 bidirectional origin (SV40) |
| Host range: | Escherichia coli; preferably recombination-deficient and endonuclease A deficient strains Mammalian cells; SV40 permissive cells |
| Parental clone: | pGADmBclxL; pEF6/E-Myc-HisA |
| Further information: | The plasmid was constructed by inserting the mouse BclxL cDNA, isolated from pGADmBclxL by the use of primers, as a SpeI/XbaI fragment between the SpeI and XbaI sites of pEF6/E-Myc-HisA. As a result, mBclxL was fused in phase to the C-terminal Myc epitope and His-tag, and the E-tag was eliminated. The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727079.1. The nucleotide sequence of the mBclxL cDNA corresponds with the EMBL Nucleotide Sequence database accession number U51278.1. Other name of the plasmid is pEF6mBclxLHis-myc. |
| EMBL Accession number: | U51278.1, view at EMBL, GenBank, DDBJ LT727079.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 20/11/2003 |
| Sequence detail: | Forward primer: 5' GGACTAGTAAGCGGCCGCAATGGCTAGC.ATG.TCT.CAG.AGC.AAC.CGG.GAG 3'
SpeI NotI
Reverse primer: 5' ATAGTTTAGCGGCCGC.TCA.TCT.AGA.CTT.CCG.ACT.GAA.GAG.TGA.GC 3'
NotI XbaI
Nucleotide sequence at the 3' end of the mouse BclxL coding sequence:
----------------- mouse BclxL ----------------->
224 229 233
5' ... GGT.GTG.GTT.CTG.CTG.GGC.TCA.CTC.TTC.AGT.CGG.AAG.TCT.AGA.GGG.CCC.TTC.
Gly Val Val Leu Leu Gly Ser Leu Phe Ser Arg Lys Ser Arg Gly Pro Phe
XbaI AsuII
------------- Myc epitope ------------> ------ His-
GAA.CAA.AAA.CTC.ATC.TCA.GAA.GAG.GAT.CTG.AAT.ATG.CAT.ACC.GGT.CAT.CAT.CAC
Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Asn Met His Thr Gly His His His
tag ------>
CAT.CAC.CAT.TGA.GTTTAAACCCGCTGATCAGCCTCGA ... 3'
His His His +++ PmeI
+++: termination codon.
Punctuation indicates reading frame. |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BamHI, BglII/NheI, HindII, NotI, SalI and SpeI/XbaI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by M. Van Gurp(1) (2) and Prof. Dr P. Vandenabeele(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: | LMBP 04772 |
Refer in your Materials and Methods: |
pEF6mBclxLMycHis (LMBP 5078) is available at BCCM/GeneCorner. This plasmid was deposited by M. Van Gurp and 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.