Last data update: 22 October 2020 04:25 CEST
Plasmid name: pCD-LCK-HLH-F-MALT1C-GyrB (LMBP 6052)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner non-core plasmid|
|Cloned DNA:||Human MALT1 paracaspase cDNA (MALT1, MLT, PCASP1, GeneID 10892); fragment
Escherichia coli DNA gyrase subunit B; fragment
Mouse lymphocyte protein tyrosine kinase cDNA (Lck); myristoylation-targeting sequence, N-terminal
HLH dimerization domain; N-terminal
FLAG epitope tag; N-terminal
|Promoter:||Human cytomegalovirus immediate early promoter (CMV-IE) and enhancer
Phage T7 gene 10 promoter (T7g10)
Simian virus 40 early promoter (SV40 early)
Escherichia coli lac operon promoter
|Terminator:||Bovine growth hormone polyadenylation signal (BGH polyA)
Simian virus 40 polyadenylation signal (SV40 polyA)
|Selection marker:||Ampicillin (amp)
Neomycin (neo; G418)
|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:||pCAGGS-E-MALT1C::GyrB; pCD-F-hSerpinB6|
|Further information:||The plasmid was created by inserting the PCR amplified HLH domain and LCK tag sequence as HindIII/KpnI fragments into pCD-F-hSerpinB6. The human SERPINB6 coding sequence was subsequently cut out with KpnI/ApaI and replaced by the minimal active domain of the human MALT1 coding sequence from pCAGGS-E-MALT1C::GyrB.
The plasmid contains the N-terminal fragment of E. coli GyrB, consisting of amino acids 2 up to and including 220, fused to a C-terminal fragment of human MALT1, consisting of codons 334 up to and including 824
This plasmid is used for mapping the minimal active domain of MALT1 for proteolytic activity.
The parental clone of this vector has been shown to provide resistance to kanamycin, although growth was reduced compared to growth on medium containing ampicillin.
The nucleotide sequence of the 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:||09/04/2009|
Primers used to amplify the minimal active domain of MALT1: KpnI-MALT1C-F: 5' atggtaccaacaactgaccagcctttggcga ApaI-GyrB-R: 5' ggcGGGCCCGCCTTCATAGTGGAAGTGGTCTT
|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) UGent-VIB Center for Inflammation Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
|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:||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)|
|Other culture collection numbers:||-|
Refer in your Materials and Methods:
|pCD-LCK-HLH-F-MALT1C-GyrB (LMBP 6052) is available at BCCM/GeneCorner. This plasmid was deposited by Dr J. Staal and 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.