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GeneCorner plasmid details

Last data update: 24 January 2024 16:39 CET

Plasmid name: Cre excised RMCE-hA20-ZF4mut (LMBP 9062)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p9062.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Human TNF alpha induced protein 3 cDNA (TNFAIP3, A20, OTUD7C, GeneID 7128); mutant
Promoter: Mouse phosphoglycerate kinase 1 promoter (PGK1)
Escherichia coli lac operon promoter; mutant (lacUV5)
Escherichia coli lac operon promoter
Ribosome
binding site:
Internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV) polyprotein gene
Terminator: Bovine growth hormone polyadenylation signal (BGH polyA)
Selection marker: Ampicillin (amp)
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli
Mammalian cells
Parental clone: Cre excised pRMCE DV1; pENTR-hA20-ZF4mut
Further information: The plasmid was constructed via LR reaction between Cre excised pRMCE DV1 and pENTR-hA20-ZF4mut.
The E-tagged mutant human A20 gene contains the kozak sequence.
The hA20 insert contains 2 point mutations (C624A and C627A) in the catalytic site of Zinc Finger 4 in order to abolish the ubiquitination activity of TNFAIP3 on RIPK1.
The Cre excision was done in vitro therefore after recombination in the ROSA26 locus the gene is immediately expressed.
The G4 ROSALUC ES cells (LMBP 10507CB) are available at BCCM/GeneCorner as well.
EMBL Accession number: -
Latest sequence update: 31/03/2014
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 M. Farhang(1) (2) and Prof. Dr G. Van Loo(1) (2).
(1) Inflammation Research Center, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: -
Related plasmid reference: Haenebalcke et al., Stem Cell Rev. Rep. 9 (2013), 774-785 [PMID: 23877658] [DOI: 10.1007/s12015-013-9458-z]
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
Cultivation remark: -
Other culture collection numbers: -

Refer in your Materials and Methods:

Cre excised RMCE-hA20-ZF4mut (LMBP 9062) is available at BCCM/GeneCorner. This plasmid was deposited by Dr M. Farhang and Prof. Dr G. Van Loo .

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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