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GeneCorner plasmid details

Last data update: 26 October 2020 04:25 CET

Plasmid name: pCD-F-NvCARD-CC (LMBP 9854)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p9854.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Nematostella vectensis caspase recruitment domain homolog cDNA (CARD-CC)
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)
Host range: Escherichia coli
Mammalian cells; SV40 permissive cells
Parental clone: pCD-F
Further information: The plasmid was constructed by PCR amplifying the Nematostella CARD-CC coding sequence, and cloning it into the KpnI/ApaI opened pCD-F vector.
Vertebrates contain 4 CARD-coiled-coil (CC) proteins that can form a CARD-CC/Bcl10/MALT1 (CBM) complex: CARD9, 10, 11 and 14.
Invertebrates that have a Bcl10 homolog typically also have a CARD-CC homolog. The Nematostella homolog represents the evolutionary most distant homologs that currently can be identified.
Other name of the plasmid is pCD-F-NvCARMA.
EMBL Accession number: -
Latest sequence update: 01/10/2018
Sequence detail:
Primers used to amplify the Nematostella CARD-CC coding sequence:

Forward: 5' ATAAGGTACCA.ATG.GAG.AAC.GAG.GAA.GAT.TCA.GA
                KpnI    ***

Reverse: 5' AATGAGGGCCC.TCA.GCT.TGT.CTT.CGC.CAG.GTT
                 ApaI   +++

***: Starting codon
+++: Termination codon
Punctuation indicates reading frame.
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: ApaI/KpnI, BglI, BglII, NcoI and PstI.
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) VIB-UGent Center for Inflammation Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: Staal et al., bioRxiv (2016) [DOI: 10.1101/046789]
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: Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051]
Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187]
Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791]
Rodriguez-Quinones et al., Focus 15 (1993), 110-112
Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660]
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:

pCD-F-NvCARD-CC (LMBP 9854) is available at BCCM/GeneCorner. This plasmid was deposited by Dr J. Staal and Prof. Dr R. Beyaert and was published in Staal et al., 2016.

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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