Last data update: 24 January 2024 16:39 CET
Plasmid name: pEF6-eIF2-A-Myc-His (LMBP 4506)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p4506.gb
(View with Genome Compiler) p4506.txt p4506.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Eukaryotic translation initiation factor 2 cDNA (eIF2); α-subunit 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 α-subunit of the eIF2 cDNA was obtained by RT PCR on HeLa mRNA. This cDNA was treated with BamHI-NotI and inserted into the BamHI-NotI opened pEF6/Myc-HisA vector, in frame with a C-terminal Myc epitope and a polyhistidine tag. The eIF2-α protein is a substrate of dsRNA-activated protein kinase (PKR). The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727240.1. The nucleotide sequence of pEF6/Myc-HisA was obtained from Invitrogen (http://www.invitrogen.com; Version C). The nucleotide sequence at the 3' end of the eIF2-α insert (from nucleotide position 2529 up to and including 2840) was analysed at the Department of Biomedical Molecular Biology (Ghent University, Belgium). Name mentioned in Saelens et al. (2001) is pEF6-eIF2-α-Myc-His. Other name of the plasmid is pEF6-eIF2-alpha-Myc-His. |
| EMBL Accession number: | LT727240.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 02/09/2003 |
| Sequence detail: | Nucleotide sequence at the 3' end of the eIF2-α cDNA:
eIF2-α ---> --> His4 --> ------- Myc
5' ... GCT.GAA.GAT.CAC.CAC.CAC.CAC.CTG.AGC.GGC.CGC.GAC.TCT.AGA.GGG.CCC.TTC.GAA.CAA.AAA
Ala Glu Asp His His His His Leu Ser Gly Arg Asp Ser Arg Gly Pro Phe Glu Gln Lys
NotI XbaI ApaI AsuII
epitope ------------------> ------- His-tag ------>
CTC.ATC.TCA.GAA.GAG.GAT.CTG.AAT.ATG.CAT.ACC.GGT.CAT.CAT.CAC.CAT.CAC.CAT.TGA ... 3'
Leu Asn Ser Glu Glu Asp Leu Asn Met His Thr Gly His His His His His His ***
AgeI
***: termination codon.
Punctuation indicates reading frame. |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BalI, PvuII, SacII, XhoI, XmaI and XmnI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Dr X. Saelens(1) (2), Dr M. Kalai(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: | Saelens et al., J. Biol. Chem. 276 (2001), 41620-41628 [PMID: 11555640] |
| Related plasmid reference: | Hsiao et al., Virology 329 (2004), 199-212 [PMID: 15476887] [DOI: 10.1016/j.virol.2004.07.032] |
| 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: |
pEF6-eIF2-A-Myc-His (LMBP 4506) is available at BCCM/GeneCorner. This plasmid was deposited by Dr X. Saelens , Dr M. Kalai and Prof. Dr P. Vandenabeele and was published in Saelens et al., 2001. |
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.