Last data update: 24 January 2024 16:39 CET
Plasmid name: pEF6-P2A-AOX1a (LMBP 9326)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p9326.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Porcine teschovirus-1 2A peptide (P2A) |
| 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) |
| Selection marker: | Cyanide / Antimycin A (Ama); in-frame translation 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-P2A |
| Further information: | The plasmid was constructed by PCR amplifying the Arabidopsis thaliana alternative oxidase AOX1a CDS and cloning it in frame with the P2A sequence in pEF6-P2A. Arabidopsis cDNA was kindly provided by Simon Stael (PSB, VIB/UGent). P2A is a self-cleaving peptide, fulfilling the same role as an IRES, but with a much shorter sequence. The purpose of this plasmid is to enforce expression of your gene of interest in selection-resistant cells by expressing the antibiotic resistance marker as an in-frame translation with your protein of interest, linked by the P2A autoprocessing peptide. Overexpression of a plant alternative oxidase has been shown to reduce reactive oxygen species (ROS) production after Antimycin A or cyanide treatment (Matsukawa et al., 2009). Further optimisation might be needed to use this as a selectable marker. |
| EMBL Accession number: | - |
| Latest sequence update: | 12/08/2014 |
| 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. Staal(1) (2) and Prof. Dr R. Beyaert(1) (2). (1) Inflammation Research Center, VIB, Ghent, Belgium (2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | - |
| Related plasmid reference: | Matsukawa et al., FEBS Lett. 583 (2009), 148-152 [PMID: 19059403] |
| Restricted distribution: | - 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-P2A-AOX1a (LMBP 9326) is available at BCCM/GeneCorner. This plasmid was deposited by Dr J. Staal and Prof. Dr R. Beyaert . |
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.