Last data update: 24 January 2024 16:39 CET
Plasmid name: pCAGGS-E-hA20-C624A-C627A-C775A-C779A (LMBP 6570)
New search | Print data sheet |
Price category: | Cat. 1 (cf. price list) |
Status: | GeneCorner non-core plasmid |
Depositor's sequence: | not available |
Sequence analysis results Genecorner: |
- |
Cloned DNA: |
Human TNF alpha induced protein 3 cDNA (TNFAIP3, A20, OTUD7C, GeneID 7128); mutated sequence E-tag; N-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-hA20-C624A-C627A; pCAGGS-E-hA20-C775A-C779A |
Further information: | pCAGGS-EhA20-C624A-C627A or pCAGGS-EhA20-C775A-C779A digested with SmaI results in a fragment of 6680 and 598. The small fragment of C624A-C627A was ligated to the large fragment of C775A-C779A to obtain pCAGGS-E-hA20-C624A-C627A-C775A-C779A. This construct has 4 point mutations disrupting Zinc Finger 4 and 7. These Zinc fingers are needed for NF-kB inhibition. The plasmid is useful for highly efficient expression of the fusion gene under the control of the AG promoter and the human CMV-IE enhancer in various mammalian cells. The AG promoter sequence consists of the chicken β-actin promoter, the first exon and part of the first intron (that seems to have a strong enhancer-like activity) linked to a rabbit β-globin fragment, consisting of a 3' part of the second intron (inclusive a branch point which is required for normal splicing reactions) and a 5' part of the third exon. When cloning a fragment downstream from the lac promoter it may be advisable to use lacIq strains in order to prevent fortuitous expression of a possibly noxious polypeptide. |
EMBL Accession number: | - |
Latest sequence update: | 22/01/2010 |
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 Prof. Dr R. Beyaert(1) (2). (1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium (2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
Plasmid reference: | Verhelst et al., EMBO J. 31 (2012), 3845-3855 [PMID: 23032186] [DOI: 10.1038/emboj.2012.240] |
Related plasmid reference: | Afonina et al., EMBO Rep. 17 (2016), 914-927 [PMID: 27113748] [DOI: 10.15252/embr.201642109] Li et al., Biochim. Biophys. Acta 1793 (2009), 346-353 [PMID: 18952128] Li et al., Biochim. Biophys. Acta 1783 (2008), 1140-1149 [PMID: 18329387] Lin et al., J. Biol. Chem. 281 (2006), 2095-2130 [PMID: 16306043] Natoli et al., J. Biol. Chem. 273 (1998), 31262-31272 [PMID: 9813034] Wertz et al., Nature 430 (2004), 694-699 [PMID: 15258597] |
Restricted distribution: | - BCCM MTA |
Distributed as: | H/P active culture or plasmid DNA |
Host for distribution: | Escherichia coli K12 MC1061 |
Host reference: | Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493] |
Related host reference: | Brigé et al., Biochem. J. 394 (2006), 335-344 [PMID: 16293111] |
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: |
pCAGGS-E-hA20-C624A-C627A-C775A-C779A (LMBP 6570) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Beyaert and was published in Verhelst et al., 2012. |
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.