Last data update: 24 January 2024 16:39 CET
Plasmid name: pEF6-P2A-mcrA (LMBP 10177)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p10177.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) Simian virus 40 polyadenylation signal (SV40 polyA) |
| Selection marker: | Mitomycin C (in-frame translation) Blasticidin (bsd) Ampicillin (amp) |
| 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 Streptomyces lavendulae mcrA coding sequence from gDNA and cloning it into the NotI/PmeI opened pEF6-P2A. The mitomycin C resistance gene from Streptomyces lavendulae can be used as a resistance marker in mammalian cells. 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. |
| EMBL Accession number: | - |
| Latest sequence update: | 05/12/2016 |
| Authenticity test: | The plasmid still needs to be subjected to the authenticity test. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | The plasmid was deposited by Dr J. Staal(1)(2) and Prof. Dr R. Beyaert(1)(2). (1) Center for Inflammation Research, VIB, Ghent, Belgium (2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | - |
| Related plasmid reference: | Belcourt et al., Proc. Natl. Acad. Sci. U.S.A. 96 (1999), 10489-10494 [PMID: 10468636] [DOI: 10.1073/pnas.96.18.10489] |
| Restricted distribution: | - BCCM MTA - The depositor will be informed of the customer's identity upon release of a sample outside the depositor's department or outside the departments in which BCCM/GeneCorner is embedded, namely UGent-DBMB and VIB-IRC. |
| 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 + kanamycin (50 μg/ml) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pEF6-P2A-mcrA (LMBP 10177) is available at BCCM/GeneCorner. The 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.