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GeneCorner plasmid details

Last data update: 21 October 2020 04:26 CEST

Plasmid name: pCAGGS-E-hRip2-CARD (LMBP 6120)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: not available
Sequence
analysis results
Genecorner:

-

Cloned DNA: Human receptor-interacting serine-threonine kinase 2 cDNA (RIP2, RIPK2, CARD3, CARDIAK, RICK); caspase-recruitment domain (CARD)
E-tag; N-terminal
Promoter: Chicken β-actin/rabbit β-globin hybrid promoter (AG)
Human cytomegalovirus immediate early promoter (CMV-IE); enhancer only
Escherichia coli lac operon promoter
Ribosome
binding site:
-
Terminator: Rabbit β-globin polyadenylation signal (β-globin polyA)
Simian virus 40 polyadenylation signal (SV40 polyA)
Selection marker: Ampicillin (amp)
Replicon: Escherichia coli plasmid pMB1 origin
Simian virus 40 bidirectional origin (SV40)
Host range: Escherichia coli
Mammalian cells; SV40 permissive cells
Parental clone: pCAGGS
Further information: pCAGGS-E-hRip2-CARD is useful for highly efficient expression of the CARD domain of human RIP2 under the control of the AG promoter and the hCMV-IE enhancer in various mammalian cells.
The dominant-negative character of the plasmid doesn't seem to be functional: inability to interfere with known RIP2- dependent NF-κB activating pathways (eg. NF-κB activation due to overexpression of RIP2 in HEK293t could not be blocked) (Kristof Kersse: personal communication).
When cloning a fragment downstream from the lac promoter it may be advisable to use lacI(q) strains in order to prevent fortuitous expression of a possibly noxious polypeptide.
EMBL Accession number: AF078530, view at EMBL, GenBank, DDBJ
Latest sequence update: 05/02/2009
Authenticity test: The plasmid still needs to be subjected to the authenticity test.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by K. Kersse and Prof. Dr P. Vandenabeele(1).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: -
Related plasmid reference: Lamkanfi et al., J. Biol. Chem. 279 (2004), 24785-24793 [PMID: 15039421]
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 MC1061
Host reference: Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493]
Related host reference: Brigé et al., Biochem. J. 394 (2006), 335-344 [PMID: 16293111]
Cultivation medium: LB-Lennox + ampicillin (100 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
Cultivation remark: -
Other culture collection numbers: -

Refer in your Materials and Methods:

pCAGGS-E-hRip2-CARD (LMBP 6120) is available at BCCM/GeneCorner. This plasmid was deposited by K. Kersse and 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.

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