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GeneCorner plasmid details

Last data update: 26 October 2020 04:25 CET

Plasmid name: pCAGGS-E-CFP-A20-YFP (LMBP 6024)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p6024.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Human TNF alpha induced protein 3 cDNA (TNFAIP3, A20, OTUD7C, GeneID 7128)
E-tag; N-terminal
Aequorea victoria green fluorescent protein DNA (GFP); enhanced cyan fluorescent variant (ECFP), N-terminal
Aequorea victoria green fluorescent protein DNA (GFP); enhanced yellow-green fluorescent variant (EYFP), C-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-E-CFP-A20; pCAGGS-E-A20-YFP
Further information: The plasmid was constructed by isolating a C-terminal fragment of the human TNFAIP3 coding sequence along with the EYFP coding sequence from pCAGGS-E-A20-YFP. This region was subsequently cloned into the BstXI opened pCAGGS-E-CFP-A20 vector.
The plasmid was created in order to monitor MALT1 proteolytic activity. After cleavage by MALT1, the C-terminal part of TNFAIP3 is rapidly degraded. Measurement of the CFP/YFP ratio results in a good in-cell quantitative measure of MALT1 activity for 1) localization of TNFAIP3 cleavage and 2) high-throughput assay for proteolytic inhibitor screenings.
The nucleotide sequence of the human TNFAIP3 cDNA corresponds with the EMBL Nucleotide Sequence Database accession number M59465.1.
EMBL Accession number: M59465.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 10/03/2009
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 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: -
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:

pCAGGS-E-CFP-A20-YFP (LMBP 6024) 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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