Last data update: 24 January 2024 16:39 CET
Plasmid name: pEF6-P2A-Hyg (LMBP 9215)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p9215.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) Escherichia coli lac operon promoter |
| Ribosome binding site: |
- |
| Terminator: | Simian virus 40 polyadenylation signal (SV40 polyA) |
| Selection marker: | Ampicillin (amp) Hygromycin (hyg); in-frame translation |
| Replicon: | Escherichia coli plasmid pMB1 origin |
| 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 hygromycin resistance CDS and cloning it into the NotI/PmeI opened pEF6-P2A. P2A is a self-cleaving peptide, fulfilling the same role as an IRES, but has 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 gene of interest, linked by the P2A autoprocessing peptide. Part of the pEF6 backbone was lost during construction, resulting in a deletion of among others the SV40 origin of replication. As a result, this plasmid is only suitable for stable expression after random genomic integration of the linearised plasmid. |
| EMBL Accession number: | - |
| Latest sequence update: | 28/11/2014 |
| 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: | - |
| 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-Hyg (LMBP 9215) 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.