Last data update: 24 January 2024 16:39 CET
Plasmid name: pdcDNA3-Myr-FLAG-hCARD9-L213LI (LMBP 10189)
New search | Print data sheet |
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 Mouse lymphocyte protein tyrosine kinase cDNA (Lck); myristoylation-targeting sequence, N-terminal 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: | 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: | pENTR3C-hCARD9-L213LI; pDEST-myr |
Further information: | This Gateway expression vector was constructed by cloning the mutated human CARD9 coding sequence from pENTR3C-hCARD9-L213LI into the pDEST-myr vector via Gateway LR recombination. The L/LI mutation corresponds to the oncogenic L225LI mutation in human CARD11. Upon overexpression, human CARD9 shows very low activation of MALT1-dependent protease and NF-kB. This construct is made to study activated human CARD9 by increasing the activity 8-fold. The purpose of this construct is to investigate whether lipid raft localization of human CARD9 will cause it to be as efficient as the CARMA proteins in NF-kB activation. One hypothesis says that the MAGUK domains in the CARMA proteins recruit them to the membrane. This construct will add an N-terminal lipid. This might not be ideal since downstream signaling occurs via the N-terminal CARD domain. CARMA proteins supposedly bind the membrane with their C-terminal domains. Other name of the plasmid is pdCDNA3-Myr-Flag-hCARD9-L213LI. |
EMBL Accession number: | - |
Latest sequence update: | 29/10/2019 |
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 Prof. Dr R. Beyaert(1) (2) and Dr J. Staal(1) (2). (1) Center for Inflammation Research, VIB, Ghent, Belgium (2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
Plasmid reference: | - |
Related plasmid reference: | De Bruyne et al., Front. Immunol. 9 (2018), 2366 [PMID: 30429846] [DOI: 10.3389/fimmu.2018.02366] |
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: |
pdcDNA3-Myr-FLAG-hCARD9-L213LI (LMBP 10189) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Beyaert and Dr J. Staal . |
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.