Last data update: 24 January 2024 16:39 CET
Plasmid name: pcDNAhRAIDD-E (LMBP 3851)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p3851.gb
(View with Genome Compiler) p3851.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human death adaptor molecule cDNA (RAIDD, PRIDD, CRADD) E-tag; C-terminal |
| Promoter: | Human cytomegalovirus immediate early promoter (CMV-IE) and enhancer Phage T7 gene 10 promoter (T7g10) Phage SP6 promoter |
| Ribosome binding site: |
- |
| Terminator: | Simian virus 40 polyadenylation signal (SV40 polyA) |
| Selection marker: | Suppressor tRNA gene (supF; requires a p3 containing host) |
| Replicon: | Bacteriophage M13 origin Escherichia coli plasmid pMB1 origin Polyoma virus origin Simian virus 40 bidirectional origin (SV40) |
| Host range: | Escherichia coli; use strains with the p3 helper plasmid (kan resistant, amber codon in amp and tet) Mammalian cells; SV40 permissive cells |
| Parental clone: | pcDNA1-hFADD; pCRhRAIDD |
| Further information: | The plasmid was constructed by inserting the HindIII/NotI fragment of pCRhRAIDD, containing the hRAIDD coding sequence, between the HindIII and NotI sites of pcDNA1-hTRADD. pcDNAhRAIDD-E is a shuttle vector designed for stable or high-level transient expression of hRAIDD in mammalian cells. The inserted gene is expressed under control of the human CMV promoter and the SV40 polyadenylation site. A small HIV (human immunodeficiency virus) fragment (29 nucleotides) between the CMV and T7 promoter sequences has an enhancer function. The T7 and SP6 promoters can be used for in vitro transcription of the cDNA insert. The presence of the polyoma and SV40 origins of replication enables the vector to replicate episomally in mammalian cells expressing large T-antigen (e.g. WOB, COS cells). The supF tRNA suppressor gene is used for selection in E. coli MC1061[p3]. It is recommended to select on both ampicillin and tetracycline containing media. The M13 origin is useful for synthesis of single-stranded DNA upon infection with the M13 helper phage. The hRAIDD sequence corresponds with the EMBL Nucleotide Sequence Database accession number U79115 (version SV 1). The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727273.1. Other name of the plasmid is pcdnaRAIDDEtag#5. |
| EMBL Accession number: | U79115, view at EMBL, GenBank, DDBJ LT727273.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 01/10/2003 |
| Sequence detail: | Nucleotide sequence at the 3' end of the human death adaptor molecule (hRAIDD) coding sequence:
hRAIDD ->|
199| |<- E-tag
5' … TTG.GAG.TGC.GCA.GCG.GCC.GCA.GGT.GCG.CCG.GTG.CCG.TAT.CCG.GAT.CCG.
Leu Glu Cys Ala Ala Ala Ala Gly Ala Pro Val Pro Tyr Pro Asp Pro
NotI BamHI
E-tag ->|
CTG.GAA.CCG.CGT.GCC.GCA.TAG 3'
Leu Glu Pro Arg Ala Ala +++
+++: Termination codon.
Punctuation indicates reading frame. |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BamHI, HindII and HindIII. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Prof. Dr P. Vandenabeele(1)(2). (1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium (2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | - |
| Related plasmid reference: | Duan et al., Nature 385 (1997), 86-89 [PMID: 8985253] |
| Restricted distribution: | - BCCM MTA |
| Distributed as: | H/P active culture or plasmid DNA |
| Host for distribution: | Escherichia coli K12 MC1061[p3] |
| Host reference: | - |
| Related host reference: | Instruction Manual (Invitrogen) Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493] |
| Helper plasmid: | p3 |
| Cultivation medium: | LB-Lennox + ampicillin (50 μg/ml) + kanamycin (50 μg/ml) + tetracycline (10 μg/ml) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pcDNAhRAIDD-E (LMBP 3851) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr P. Vandenabeele. |
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.