Last data update: 24 January 2024 16:39 CET
Plasmid name: pEASY-FLIRT-mCYLD-R321A-KI (LMBP 7780)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p7780.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Mouse CYLD lysine 63 deubiquitinase cDNA (Cyld, CYLD1); mutated fragment |
| Promoter: | Escherichia coli lac operon promoter Phage T7 gene 10 promoter (T7g10) Herpes simplex virus (HSV) thymidine kinase promoter (TK) Mouse angiotensin I converting enzyme promoter (ACE) |
| Ribosome binding site: |
- |
| Terminator: | Bovine growth hormone polyadenylation signal (BGH polyA) |
| Selection marker: | Ampicillin (amp) Neomycin (neo; G418; kanamycin (kan)) |
| Replicon: | Escherichia coli plasmid pMB1 origin Phage f1 origin |
| Host range: | Escherichia coli Mammalian cells |
| Parental clone: | pEASY-FLIRT-mCYLD-LAH-3K; pEASY-FLIRT-mCYLD-RAH-R321A |
| Further information: | The plasmid was constructed by restriction digest of pEASY-FLIRT-mCYLD-LAH-3K and pEASY-FLIRT-mCYLD-RAH-R321A with NotI/SalI, isolating the fragments containing the mouse CYLD homology arms of both parental vectors and ligating them together. This is a knock-in construct in order to generate a MALT1-resistant CYLD mouse. The MALT1 cleavage site was eliminated via a R321A mutation. The mouse CYLD coding sequence also contains a NaeI site, introduced via a silent mutation. The plasmid contains two bacteriophage P1 Cre recombinase target sites (loxP sites) for excision of the cassette by Cre-recombinase. pEASY-FLIRT contains a testis-specific Flp expression cassette (Ace-Flpe) aimed at deleting the frt-flanked NeoR gene in male gonads of chimeric mice. Other name of the plasmid is pEASY-FLIRT-mCYLD-R321A. |
| EMBL Accession number: | - |
| Latest sequence update: | 11/06/2015 |
| 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) Inflammation Research Center, VIB, Ghent, Belgium (2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | - |
| Restricted distribution: | - VIB/BCCM MTA - Restricted to academic users |
| 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: |
pEASY-FLIRT-mCYLD-R321A-KI (LMBP 7780) 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.