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GeneCorner plasmid details

Last data update: 24 January 2024 16:39 CET

Plasmid name: pcDNA3.1-FLAG-hCARD9-T231E (LMBP 10531)

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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 caspase recruitment domain family member 9 cDNA (CARD9, GeneID 64170); mutated sequence
FLAG epitope tag; N-terminal
Promoter: Human cytomegalovirus immediate early promoter (CMV-IE) and enhancer
Phage T7 gene 10 promoter (T7g10)
Simian virus 40 early promoter (SV40 early)
Escherichia coli lac operon promoter
Ribosome
binding site:
-
Terminator: Bovine growth hormone polyadenylation signal (BGH polyA)
Simian virus 40 polyadenylation signal (SV40 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: pcDNA3.1-FLAG-hCARD9
Further information: The plasmid was created by introducing a T231E mutation into the human CARD9 coding sequence of the pcDNA3.1-FLAG-hCARD9 vector.
Human CARD9 is activated by phosphorylation by PRKCD at T231 (Strasser et al., 2012). This is a phospho-mimic mutant for constituitive activation.
The nucleotide sequence of the wild type human CARD9 coding sequence corresponds with the EMBL Nucleotide Sequence Database accession number AF311287.1.
Other name of the plasmid is pCDNA3-Flag-hCARD9-T231E.
EMBL Accession number: AF311287.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 20/04/2020
Sequence detail:
Phospho-mimic mutation was made with the primers:
    QC-hCARD9-T231E-F : tggagcgcaagcacgaactgaagctcaggcacg
    QC-hCARD9-T231E-R : cgtgcctgagcttcagttcgtgcttgcgctcca
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 Dr J. Staal(1) (2) and Prof. Dr R. Beyaert(1) (2).
(1) UGent-VIB Center for Inflammation Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: -
Related plasmid reference: Strasser et al., Immunity 36 (2012), 32-42 [PMID: 22265677] [DOI: 10.1016/j.immuni.2011.11.015]
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:

pcDNA3.1-FLAG-hCARD9-T231E (LMBP 10531) is available at BCCM/GeneCorner. This 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.

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