Last data update: 24 January 2024 16:39 CET
Plasmid name: pcDNA3.1-FLAG-hCARD9 (LMBP 9609)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p9609.gb
(View with Genome Compiler) p9609.txt p9609.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human caspase recruitment domain family member 9 cDNA (CARD9, GeneID 64170) FLAG epitope tag; N-terminal |
| Promoter: | Human cytomegalovirus immediate early promoter (CMV-IE) and enhancer Simian virus 40 early promoter (SV40 early) Phage T7 gene 10 promoter (T7g10) 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) |
| Copy number: | High copy number |
| Host range: | Escherichia coli Mammalian cells; SV40 permissive cells |
| Parental clone: | pcDNA3.1-Flag |
| Further information: | The plasmid was constructed by PCR amplifying the human CARD9 coding sequence and cloning it into the BamHI/EcoRV opened pcDNA3.1-Flag vector. The plasmid is intended for eukaryotic expression of the human CARD9 protein. 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 the human CARD9 coding sequence corresponds with Genbank accession number NM_052813.5. The nucleotide sequence of the pcDNA3.1-Flag plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727011.1. Other name of the plasmid is pCDNA3-Flag-hCARD9 |
| EMBL Accession number: | NM_052813.5, view at GenBank LT727011.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 04/08/2020 |
| Sequence detail: | Primers used to amplify the human CARD9 coding sequence:
Forward: 5' CTAAGGATCCA.ATG.TCG.GAC.TAC.GAG.AAC.GAT.GA
BamHI ***
Reverse: 5' CTA.GGA.GCC.CTC.AGT.GTC.GGT.GTT
+++
***: starting codon
+++: termination codon
Punctuation indicates reading frame. |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: EcoRV/KpnI, NcoI, PvuI and XhoI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Dr J. Staal(1) (2) and Prof. R. Beyaert(1) (2). (1) VIB-UGent Center for Inflammation Research, VIB, Ghent, Belgium (2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | De Bruyne et al., Front. Immunol. 9 (2018), 2366 [PMID: 30429846] [DOI: 10.3389/fimmu.2018.02366] |
| Related plasmid reference: | Staal et al., bioRxiv (2016), 1-42 [DOI: 10.1101/046789] Staal et al., bioRxiv (2017), 1-25 [DOI: 10.1101/228767] Afonina et al., EMBO Rep. 17 (2016), 914-927 [PMID: 27113748] [DOI: 10.15252/embr.201642109] |
| 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 (LMBP 9609) is available at BCCM/GeneCorner. This plasmid was deposited by Dr J. Staal and Prof. R. Beyaert and was published in De Bruyne et al., 2018. |
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.