Last data update: 24 January 2024 16:39 CET
Plasmid name: pOPINE-HA-hDRD2-BRIL-Avi-His (LMBP 10445)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p10445.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Influenza A virus hemagglutinin cDNA (HA); signal sequence Human dopamine receptor D2 cDNA (DRD2, GeneID 1813); lacking intracellular loop 3 (∆ICL3) Escherichia coli cytochrome b562 cDNA (GeneID 13703733); codon-optimized fragment (BRIL) Avidin biotinylation sequence (AviTag); C-terminal Histidine tag (His-tag); C-terminal |
| Promoter: | Human cytomegalovirus immediate early promoter (CMV-IE); enhancer only Chicken β-actin promoter (ACTB) Phage T7 gene 10 promoter (T7g10) with lac operator (T7lac) Autographa californica nuclear polyhedrosis virus (AcNPV) p10 promoter (p10) |
| Ribosome binding site: |
- |
| Terminator: | Rabbit β-globin polyadenylation signal (β-globin polyA) Phage T7 gene 10 terminator (T7g10) |
| Selection marker: | Ampicillin (amp) |
| Replicon: | Escherichia coli plasmid pMB1 origin |
| Host range: | Escherichia coli Mammalian cells Insect cells; e.g. Sf9 cells |
| Parental clone: | pOPINE |
| Further information: | The plasmid was constructed by cloning the DRD2∆ICL3-BRIL coding sequence, with the Influenza A virus HA signal sequence and a C-terminal AviTag, into the pOPINE vector. The dopamine receptor D2 belongs to the G protein-coupled receptors. This vector contains the hybrid CMV-enhancer/Chicken β-actin promoter that has been reported to give higher expression levels compared to vectors using CMV-derived promoter and enhancer. In addition this vector contains a Kozak consensus sequence for efficient initiation of translation in eukaryotic hosts. The presence of the p10 baculoviral promoter and the flanking lef2/ORF603 and ORF1629 baculoviral recombination sites allow the construction of recombinant baculoviruses and, finally, a T7 polymerase promoter with lacO operator offers high level inducible expression in E.coli harbouring the λ (DE3) prophage. The BRIL soluble domain promotes expression by stabilizing the receptor and increasing the solubility. The nucleotide sequence of the pOPINE vector corresponds with the EMBL Nucleotide Sequence Database accession number EF372397.1. Other name of the plasmid is pOPINE-HA-D2BRIL-Avi-His. |
| EMBL Accession number: | EF372397.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 16/08/2017 |
| 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 Prof. Dr S. Savvides(1)(2). It was constructed by Dr. I. Ramou(1)(2). (1) Department of Biochemistry and Microbiology, Ghent University, Ghent, Belgium (2) VIB-UGent Center for Inflammation Research, VIB, Ghent, Belgium |
| Plasmid reference: | - |
| Related plasmid reference: | Berrow et al., Nucleic Acids Res. 35 (2007), e45 [PMID: 17317681] [DOI: 10.1093/nar/gkm047] |
| 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: |
pOPINE-HA-hDRD2-BRIL-Avi-His (LMBP 10445) is available at BCCM/GeneCorner. The plasmid was deposited by Prof. Dr S. Savvides. |
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.