Last data update: 24 January 2024 16:39 CET
Plasmid name: pClRM1-T7pol (LMBP 9330)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p9330.gb
(View with Genome Compiler) p9330.txt p9330.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Phage T7 RNA polymerase gene (T7g1) SV40 large T-antigen nuclear localization signal (NLS); N-terminal Vesicular stomatitis virus glycoprotein (VSV-G) epitope; N-terminal Synthetic nuclear export signal (NES); N-terminal Influenza HA epitope encoding the haemagglutinin tagging peptide; N-terminal Mouse lymphocyte protein tyrosine kinase cDNA (Lck); myristoylation-targeting sequence, N-terminal Porcine teschovirus-1 2A peptide (P2A) |
| Promoter: | Human elongation factor 1α promoter (EF1α) Simian virus 40 early promoter (SV40 early) Escherichia coli lac operon promoter Synthetic prokaryotic EM7 promoter Phage T7 gene 10 promoter (T7g10) |
| Ribosome binding site: |
- |
| Terminator: | Bovine growth hormone polyadenylation signal (BGH polyA) Simian virus 40 polyadenylation signal (SV40 polyA) |
| Selection marker: | Blasticidin (bsd) Ampicillin (amp) |
| Replicon: | Escherichia coli plasmid pMB1 origin Phage f1 origin Simian virus 40 bidirectional origin (SV40) |
| Host range: | Escherichia coli Mammalian cells; SV40 permissive cells |
| Parental clone: | pClR-M1; pT7POL26 |
| Further information: | The plasmid was constructed by PCR amplifying the T7 RNA polymerase sequence (without stop codon) from pT7POL26 and cloning it into the BamHI/EcoRI opened pCIR-M1 vector. This vector consists of the following elements: Myr-HA-NES-MALT1 cleavage site-VSV-NLS-BamHI-T7POL26-EcoRI-P2A-EcoRV. Uncleaved, T7 RNA polymerase is kept out of the nucleus by nuclear export and membrane anchoring. Upon cleavage, T7 RNA polymerase will enter the nucleus and drive expression of a reporter cloned behind the T7 promoter. |
| EMBL Accession number: | - |
| Latest sequence update: | 15/03/2022 |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BamHI, KpnI and PvuII. |
| 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) VIB Center for Inflammation Research, 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 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 |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pClRM1-T7pol (LMBP 9330) 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.