Last data update: 24 January 2024 16:39 CET
Plasmid name: pcDNAFLAGhRAIDDMyc (LMBP 3842)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p3842.gb
(View with Genome Compiler) p3842.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human death adaptor molecule cDNA (RAIDD, PRIDD, CRADD) Myc epitope; C-terminal FLAG epitope tag; N-terminal |
| Promoter: | Human cytomegalovirus immediate early promoter (CMV-IE) and enhancer Phage T7 gene 10 promoter (T7g10) Phage SP6 promoter Simian virus 40 early promoter (SV40 early) |
| Ribosome binding site: |
- |
| Terminator: | Simian virus 40 polyadenylation signal (SV40 polyA) Bovine growth hormone polyadenylation signal (BGH polyA) |
| Selection marker: | Ampicillin (amp) Neomycin (neo; G418) |
| 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: | pcDNA3 |
| Further information: | The plasmid was constructed by inserting the human death adaptor molecule (hRAIDD) coding sequence between the HindIII and XhoI sites of pcDNA3 by the use of PCR primers, containing a C-terminal Myc epitope and N-terminal FLAG epitope tag respectively. The hRAIDD sequence corresponds with the EMBL Nucleotide Sequence Database accession number U79115 (version SV 1). There is uncertainty about the presence of a second enhancer in the SV40 early promoter. The absence of a second enhancer in this promoter may cause bacterial leakage expression of the Tn5 neomycin resistance gene. This would cause transformed E. coli cells to be resistant to kanamycin, although growth should be reduced compared to growth on medium containing ampicillin. The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727264.1. Other name of the plasmid is pcDNA3flagPRIDDmyc. |
| EMBL Accession number: | U79115, view at EMBL, GenBank, DDBJ LT727264.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 30/09/2003 |
| Sequence detail: | Sequence at the 5' border of the human death adaptor molecule (hRAIDD) coding sequence:
<--------- FLAG tag ----------> <------- hRAIDD
5' AAGCTTACCACC.ATG.GAC.TAC.AAG.GAT.GAC.GAT.GAC.AAA.GCG.GCC.GCA.ATG.GAG.GCC.AGA 3'
HindIII Met Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ala Ala Met Glu Ala Arg
NcoI NotI
Sequence at the 3' border of the human death adaptor molecule (hRAIDD) coding sequence:
hRAIDD----> <------------ Myc epitope ------------>
5' ATG.TTG.GAG.GCT.AGC.GAA.CAG.AAG.CTG.ATA.TCG.GAA.GAG.GAC.CTG.AAC.TAG.TAACTCGAG 3'
Met Leu Glu Ala Ser Glu Glu Lys Leu Ile Ser Glu Glu Asp Leu Asn * XhoI |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: HindIII, ScaI and XhoI. |
| 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(λ) |
| Host reference: | Mertens et al., Gene 164 (1995), 9-15 [PMID: 7590329] |
| Related host reference: | Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493] |
| Cultivation medium: | LB-Lennox + ampicillin (100 μg/ml) |
| Cultivation temperature: | 28°C |
| Biosafety level: | L1 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pcDNAFLAGhRAIDDMyc (LMBP 3842) 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.