Last data update: 24 January 2024 16:39 CET
Plasmid name: pEF6-hbeclin-1-D133AD149A (LMBP 7301)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p7301.gb
(View with Genome Compiler) p7301.txt p7301.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human beclin 1 cDNA (BECN1, ATG6, VPS30); mutated sequence 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: | pEF6/Myc-HisA |
| Further information: | The plasmid was constructed by ligating the PCR amplified human BECN1 cDNA as a KpnI/NotI fragment into the KpnI/NotI opened pEF6/Myc-HisA vector, and inserting D133A and D149A mutations via site-directed PCR mutagenesis. The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT726855.1. The nucleotide sequence of the human BECN1 cDNA corresponds with the EMBL Nucleotide Sequence Database accession number AF077301.1, except for the inserted D133A and D149A mutations and the A nucleotide at position 1164 of the full coding sequence, which was analysed by the depositor as a T, resulting in a silent point mutation. By lack of construction data, the nucleotide sequence at the C-terminal end of the human BECN1 coding sequence could not be completely traced back. However, as the depositor confirmed that the BECN1 amplicon was cloned in frame with the Myc epitope and His-tag in pEF6-hbeclin-1-Myc-His (LMBP 7300), BCCM/GeneCorner assumes that this is also the case in pEF6-hbeclin-1-D133AD149A. Other names of the plasmid are pEF6-Beclin-D133A-D149A and pEF6-Beclin-DQLD-VD-Hismyc. |
| EMBL Accession number: | AF077301.1, view at EMBL, GenBank, DDBJ LT726855.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 07/07/2011 |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BalI, BglII, KpnI/NotI, SacI and XmnI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by E. Wirawan(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: | Wirawan et al., Cell Death Dis. 1 (2010), e18 [PMID: 21364619] [DOI: 10.1038/cddis.2009.16] |
| Restricted distribution: | - VIB/BCCM MTA |
| Distributed as: | H/P active culture or plasmid DNA |
| Host for distribution: | Escherichia coli K12 MC1061 |
| Host reference: | Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493] |
| Related host reference: | Brigé et al., Biochem. J. 394 (2006), 335-344 [PMID: 16293111] |
| 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: |
pEF6-hbeclin-1-D133AD149A (LMBP 7301) is available at BCCM/GeneCorner. This plasmid was deposited by E. Wirawan and Prof. Dr P. Vandenabeele and was published in Wirawan et al., 2010. |
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.