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LMBP plasmid details

Last data update: 26 September 2020 04:22 CEST

Plasmid name: pLenti6-hPKCeta-VSV-puro (LMBP 9979)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: -
Sequence
analysis results
Genecorner:

-

Cloned DNA: Human protein kinase C eta cDNA (PRKCH, PRKCL, PKCL, PKCeta, GeneID 5583)
Vesicular stomatitis virus glycoprotein (VSV-G) epitope; C-terminal
Promoter: Human cytomegalovirus immediate early promoter (CMV-IE) and enhancer
Rous sarcoma virus/human immunodeficiency virus hybrid long terminal repeat (RSV/HIV-1 5' LTR)
Simian virus 40 early promoter (SV40 early)
Phage T3 promoter
Phage T7 gene 10 promoter (T7g10)
Escherichia coli lac operon promoter
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-VSV puro; pENTR3C-hPRKCH-no_stop
Further information: The plasmid was constructed by cloning the human PRKCH coding sequence from pENTR3C-hPRKCH-no_stop into pLenti6-VSV puro via Gateway LR recombination.
This is a lentiviral Gateway expression vector.
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 human PRKCH coding sequence corresponds with the EMBL Nucleotide Sequence Database accession number M55284.1.
The nucleotide sequence of the pLenti6-VSV puro vector corresponds with the EMBL Nucleotide Sequence Database accession number LT009459.1.
Other name of the plasmid is pLenti6puroVSV_hPKCeta wt.
EMBL Accession number: M55284.1, view at EMBL, GenBank, DDBJ
LT009459.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 13/10/2016
Authenticity test: The plasmid still needs to be subjected to the authenticity test.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: The plasmid was deposited by Prof. Dr W. Declercq(1)(2). It was constructed by K. Leurs(1)(2).
(1) Inflammation Research Center, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: PhD thesis Corinne Urwyler
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
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.

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