Last data update: 26 September 2020 04:22 CEST
Plasmid name: pLVX-EF1alpha-hMALT1-C464A-IRES-ZsGreen1 (LMBP 8928)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner non-core plasmid|
|Depositor's sequence:||not available|
|Cloned DNA:||Human MALT1 paracaspase cDNA (MALT1, MLT, PCASP1, paracaspase 1); mutated sequence
Zoanthus sp. green fluorescent protein cDNA (ZsGreen); human-codon-optimised variant (ZsGreen1)
|Promoter:||Human immunodeficiency virus long terminal repeat (HIV-1 5' LTR)
Human elongation factor 1α promoter (EF1α)
Escherichia coli lac operon promoter
|Internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV) polyprotein gene|
|Terminator:||Human immunodeficiency virus long terminal repeat (HIV-1 3' LTR); with deleted U3 region (ΔU3/3' LTR)
Simian virus 40 polyadenylation signal (SV40 polyA)
|Selection marker:||Ampicillin (amp)|
|Replicon:||Escherichia coli plasmid pMB1 origin|
|Copy number:||High copy number|
|Host range:||Escherichia coli
|Parental clone:||pLVX-EF1alpha-IRES-ZsGreen1; pCD-MALT1-C464A|
|Further information:||This lentiviral expression vector was constructed by PCR amplifying the human MALT1 coding sequence from pCD-MALT1-C464A and cloning it as a NotI/BamHI fragment into the pLVX-EF1alpha-IRES-ZsGreen1 vector.
pLVX-EF1alpha-hMALT1-C464A-IRES-ZsGreen1 is a HIV-1-based, lentiviral expression vector designed to simultaneously and constitutively express the hMALT1 C464A mutant and ZsGreen1 from a bicistronic transcript in mammalian cells.
ZsGreen1 is a human-codon-optimised variant of the reef coral Zoanthus sp. green fluorescent protein (ZsGreen) that has been engineered for brighter fluorescence. The excitation and emission maxima of native ZsGreen1 are 493 nm and 505 nm, respectively.
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 RNA, leading to increased viral titers from packaging 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.
The nucleotide sequence of the wild type human MALT1 coding sequence corresponds with the EMBL Nucleotide Sequence Database accession number AF130356.2.
|EMBL Accession number:||AF130356.2, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||13/07/2016|
|Authenticity test:||The plasmid still needs to be subjected to the authenticity test.|
|History of deposit:||This plasmid was deposited by Dr J. Staal(1) (2) and Prof. Dr R. Beyaert(1) (2).
(1) Inflammation Research Center, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
|Restricted distribution:||- VIB/BCCM MTA
- Restricted to academic users
|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:||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)|
|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.