Last data update:
20 October 2018 04:23 CEST
| Name: | pLenti6-mBeclin1-C-Frag-147-V5-puro Add to cart |
| Accession number: | LMBP 8562 |
| Sequence file: | Depositor's sequence: p8562.gb View with Genome Compiler |
| Status: | LMBP non-core plasmid |
| Cloned DNA: | Mouse beclin 1, autophagy related cDNA (Becn1, Atg6); fragment V5 epitope; C-terminal |
| Promoter: | Rous sarcoma virus/human immunodeficiency virus hybrid long terminal repeat (RSV/HIV-1 5' LTR) Human cytomegalovirus immediate early promoter (CMV-IE) and enhancer Escherichia coli lac operon promoter Simian virus 40 early promoter (SV40 early) Phage T3 promoter Phage T7 gene 10 promoter (T7g10) |
| Ribosome binding site: |
- |
| Terminator: | Rous sarcoma virus/human immunodeficiency virus hybrid long terminal repeat (RSV/HIV-1 3' LTR); modified HIV-1 3' LTR with deleted U3 region (ΔU3/3' LTR) Simian virus 40 polyadenylation signal (SV40 polyA) |
| Selection marker: | Ampicillin (amp) Puromycin (puro) |
| Replicon: | Escherichia coli plasmid pMB1 origin Phage f1 origin Simian virus 40 bidirectional origin (SV40) |
| Host range: | Escherichia coli Mammalian cells; SV40 permissive cells |
| Parental clone: | pLenti6-V5 puro; pENTR3C-mBeclin1-C-Frag-147-stop |
| Further information: | The plasmid was constructed via Gateway LR recombination of pLenti6-V5 puro and pENTR3C-mBeclin1-C-Frag-147-stop. This is a constitutive lentiviral Gateway expression vector, expressing the C-terminal fragment of the mouse Becn1 coding sequence, starting at codon 147, fused to the V5 epitope. The U3 region of the HIV-1 3' LTR is deleted (ΔU3 or dU3) and facilitates self-inactivation of the HIV-1 5' LTR after transduction to enhance the biosafety of the vector. The element also contains a polyadenylation signal for transcription termination and polyadenylation of mRNA in transduced cells. The vector contains the psi packaging signal, allowing viral packaging, and the rev response element (RRE) from HIV-1, permitting Rev-dependent nuclear export of unspliced viral mRNA. Because of the high risk for recombination, plasmid integrity should be checked via PvuII restriction digest after each subcultivation. The nucleotide sequence of the mouse Becn1 coding sequence corresponds with the EMBL Nucleotide Sequence Database accession number BC005770.1. The nucleotide sequence of the pLenti6-V5 puro vector corresponds with the EMBL Nucleotide Sequence Database accession number LT009457.1. Other name of the plasmid is pLenti6(puro)-V5-dest-mBeclin1-C-Frag_147-stop. |
| EMBL Accession number: | BC005770.1, view at EMBL, GenBank, DDBJ LT009457.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 24/08/2016 |
| Authenticity test: | The restriction enzyme pattern has still to be analysed. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | The plasmid was deposited by Prof. Dr P. Vandenabeele(1)(2). It was constructed by I. Bruggeman(1)(2). (1) Inflammation Research Center, VIB, Ghent, Belgium (2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | - |
| Related plasmid reference: | De Groote et al., BioTechniques 60 (2016), 252-259 [PMID: 27177818] [DOI: 10.2144/000114417] |
| Restricted distribution: | - BCCM MTA |
| Distributed as: | plasmid DNA |
| Host for distribution: | Escherichia coli K12 DH5α |
| Host reference: | Focus 8 (1986), 9 |
| Related host reference: | Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051] Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187] Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791] Rodriguez-Quinones et al., Focus 15 (1993), 110-112 Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660] |
| Cultivation medium: | LB-Lennox + ampicillin (100 μg/ml) |
| Cultivation temperature: | 28°C |
| Biosafety level: | L1 in E. coli; L2 in mammalian cells |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
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.