Last data update: 24 January 2024 16:39 CET
Plasmid name: pENTR221-E-hA20-R439[P2A] (LMBP 9158)
| 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 Porcine teschovirus-1 2A peptide (P2A) |
| Promoter: | Phage T7 gene 10 promoter (T7g10) |
| Ribosome binding site: |
- |
| Terminator: | Escherichia coli rrnB operon T2 terminator Escherichia coli rrnB operon T1 terminator |
| Selection marker: | Neomycin (neo; kanamycin (kan)) |
| Replicon: | Escherichia coli plasmid pMB1 origin |
| Host range: | Escherichia coli |
| Parental clone: | pDONR221; pCAGGS-E-hA20-R439A |
| Further information: | The plasmid was contructed by fusion PCR, amplifying the first and second part of the human TNFAIP3 coding sequence from pCAGGS-E-hA20-R439A and fusing a P2A sequence between them, replacing the R439 codon. The resulting coding sequence was cloned into pDONR221 via Gateway BP recombination. P2A is a non-enzymatic autoprocessing peptide for polycistronic expression. Upon transfer of the human TNFAIP3 coding sequence to an expression vector, a constituitively cleaved TNFAIP3 protein will be expressed. The nucleotide sequence of the wild type human TNFAIP3 coding sequence corresponds with EMBL Nucleotide Sequence Database accession number M59465.1. |
| EMBL Accession number: | M59465.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 20/04/2020 |
| Sequence detail: | Primers used for fusion PCR: For P2A fusion: Forward: ggaagcggagctactaacttcagcctgctgaagcaggctggagacgtggaggagaaccctggacct Reverse: aggtccagggttctcctccacgtctccagcctgcttcagcaggctgaagttagtagctccgcttcc Left fragment: cttcagcaggctgaagttagtagctccgcttccagaggccccgagcgccatgcca Right fragment: aggctggagacgtggaggagaaccctggacctggagaagcctatgagcccttg |
| 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: | 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 + kanamycin (50 μg/ml) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pENTR221-E-hA20-R439[P2A] (LMBP 9158) 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.