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LMBP plasmid details

Last data update: 25 September 2020 04:24 CEST

Plasmid name: pLenti-CMV-HA-LZTR1-M202R-puro (LMBP 11175)

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Price category: Cat. 3 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: -
Sequence
analysis results
Genecorner:

NGS: h07-gc-dec2018.fasta

Cloned DNA: Human leucine zipper like transcription regulator 1 cDNA (LZTR1, LZTR-1, BTBD29, GeneID 8216); mutated sequence
Influenza HA epitope encoding the haemagglutinin tagging peptide; N-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)
Escherichia coli lac operon promoter
Phage T3 promoter
Phage T7 gene 10 promoter (T7g10)
Mouse phosphoglycerate kinase 1 promoter (PGK1)
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: pLenti-CMV-HA-LZTR1-puro
Further information: The plasmid was constructed by introducing an M202R mutation into the human LZTR1 coding sequence of pLenti-CMV-HA-LZTR1-puro.
The nucleotide sequence of the human LZTR1 coding sequence was altered for ease of synthesis but contains no codon changes other than M202R.
The plasmid contains all of the viral processing elements necessary for the production of replication-incompetent lentivirus, as well as elements to improve viral titer, transgene expression and overall vector function.
The woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) promotes RNA processing events and enhances nuclear export of viral and transgene RNA, leading to increased viral titers from packaging cells and enhanced expression of the mutated LZTR1 in target cells.
The plasmid includes a Rev-response element (RRE), which further increases viral titers by enhancing the transport of unspliced viral RNA out of the nucleus.
The plasmid also contains a central polypurine tract/central termination sequence element (cPPT/CTS). During target cell infection, this element creates a central DNA flap that increases nuclear import of the viral genome, resulting in improved vector integration and more efficient transduction.
Because of the high risk for recombination, this plasmid is only available under the format of isolated plasmid DNA.
Other name of the plasmid is pLenti CMV HA-LZTR1 M202R puro (1701).
EMBL Accession number: -
Latest sequence update: 25/01/2019
Authenticity test: This plasmid has been fully sequenced but the NGS sequence data still need to be implemented.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Prof. Dr Anna Sablina(1)(2).
(1) Center for Cancer Biology, VIB, Leuven, Belgium
(2) Department of Oncology, University of Leuven, Leuven, Belgium
Plasmid reference: Steklov et al., Science 362 (2018), 1177-1182 [PMID: 30442762] [DOI: 10.1126/science.aap7607]
Restricted distribution: - BCCM MTA
- The depositor will be informed of the customer's identity upon release of a sample outside the depositor's department or outside the departments in which BCCM/GeneCorner is embedded, namely UGent-DBMB and VIB-IRC.
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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