Last data update: 24 January 2024 16:39 CET
Plasmid name: pDest-LZRS-E-hA20 (LMBP 5941)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p5941.gb
(View with Genome Compiler) p5941.txt p5941.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human TNF alpha induced protein 3 cDNA (TNFAIP3, A20, OTUD7C, GeneID 7128) Epstein-Barr virus nuclear antigen 1 gene (EBNA1, EBNA-1, GeneID 3783709) Aequorea victoria green fluorescent protein DNA (GFP); enhanced red-shifted variant (EGFP) E-tag; N-terminal |
| Promoter: | Moloney mouse leukemia virus long terminal repeat (5' MMLV-LTR) |
| Ribosome binding site: |
Internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV) polyprotein gene |
| Terminator: | Simian virus 40 polyadenylation signal (SV40 polyA) Moloney mouse leukemia virus long terminal repeat (3' MMLV-LTR) |
| Selection marker: | Ampicillin (amp) Puromycin (puro) |
| Replicon: | Escherichia coli plasmid pMB1 origin Epstein-Barr virus origin of replication (oriP) |
| Host range: | Escherichia coli Mammalian cells |
| Parental clone: | pEntry-2b-E-hA20; pDEST-LZRS-ires-eGFP |
| Further information: | This retroviral Gateway expression vector was constructed by inserting the human A20 cDNA from pEntry-2b-E-hA20 into the vector pDEST-LZRS-ires-eGFP via Gateway recombination with LR clonase. The regions upstream of the 5' MMLV-LTR (position 9931 to 10976 of the plasmid sequence) and downstream from the 5' MMLV-LTR (position 11632 to 12933 of the plasmid sequence) and the region between the SV40 polyA and the puromycin coding sequence (position 3308 to 4111 of the plasmid sequence) were analysed by LMBP. The nucleotide sequence of the hA20 cDNA corresponds with the EMBL Nucleotide Sequence Database accession number M59465.1. The nucleotide sequence of the EGFP DNA corresponds with the EMBL Nucleotide Sequence Database accession number U55761.1, except for an A-G and a T-A replacement at nucleotide position 69 respectively 695 of the EGFP coding sequence, the latter replacement resulting in a L232H switch. |
| EMBL Accession number: | M59465.1, view at EMBL, GenBank, DDBJ U55761.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 08/07/2011 |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BamHI, BglII, BglII/HindIII, BstEII/NsiI, HincII, PvuI, SacI, SacII, SacII/SphI, SalI, SpeI and SphI. As compared to the compiled nucleotide sequence, the restriction site analysis pattern revealed that HincII cuts eight times instead of seven times. The 3559 bp HincII fragment is splitted into two fragments of approx. 1500 bp and approx. 2050 bp respectively. The extra HincII site is thus most probably located in or in the neighborhood of the 5' MMLV-LTR region; unfortunately, the sequence analysis didn't cover this region. The SphI site at position 7303 could not be experimentally confirmed. The compiled nucleotide sequence has not been adapted accordingly. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Prof. Dr R. Beyaert(1) (2). It was constructed by Y. Bruynooghe(1) (2). (1) Department for Molecular Biomedical 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 + ampicillin (100 μg/ml) |
| Cultivation temperature: | 28°C |
| Biosafety level: | L1 in E. coli; L2 in mammalian cells |
| Cultivation remark: | This culture grows very slow: take more than 24 hours into account for saturated growth! |
| Other culture collection numbers: | - |
| Related website: | http://www.iresite.org/IRESite_web.php?page=view&entry_id=140 |
Refer in your Materials and Methods: |
pDest-LZRS-E-hA20 (LMBP 5941) is available at BCCM/GeneCorner. This plasmid was deposited by 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.