Last data update: 23 October 2020 04:27 CEST
Plasmid name: pCD-F-HLH-MK-E4A-bio (LMBP 5683)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
|Cloned DNA:||Human MALT1 paracaspase cDNA (MALT1, MLT, PCASP1, paracaspase 1); mutated sequence with Kozak consensus sequence
FLAG epitope tag; N-terminal
HLH dimerization domain; N-terminal
Biotinylation sequence (bio-tag); C-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
|Further information:||The TRAF6 binding sites of hMLT were mutated. As compared to the wt hMLT cDNA, the glutamic acid (E) codons at positions 313, 316, 653 and 806 were replaced by alanine (A) codons, creating additional SphI, AgeI and NruI sites and abolishing an Alw44I site.
The helix-loop-helix (HLH) domain is a dimerization domain that allows proteins to form homo- and/or heterodimers.
The HLH domain consists of the N-terminal sequence of hMLT (encoding the first 42 amino acids) and a fragment of the human ets variant gene 6 (hETV6) coding sequence (encoding amino acids 27 to 144).
The bio-tag is biotinylated by cotransfection with the plasmid pMSCV-F-BirA and can be used for highly efficient purification.
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 this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT726943.1
The nucleotide sequence of the wt hMLT cDNA corresponds with the EMBL Nucleotide Sequence Database accession number AF130356.2.
Other name of the plasmid is pCD-F-HLH-MK-E4A-bioC.
|EMBL Accession number:||AF130356.2, view at EMBL, GenBank, DDBJ
LT726943.1, view at EMBL, GenBank, DDBJ
|Latest sequence update:||28/11/2007|
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: AgeI, Alw44I, Alw44I/HindIII, BstXI/PstI, EcoRI, EcoRI/NcoI, HindIII, NcoI, NruI, SphI and XhoI/XmnI.|
|History of deposit:||This plasmid was deposited by Dr M. Baens(1).
(1) Department of Human Genetics, KULeuven, Belgium
|Related plasmid reference:||Noels et al., J. Biol. Chem. 282 (2007), 10180-10189 [PMID: 17287209]
|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-F-HLH-MK-E4A-bio (LMBP 5683) is available at BCCM/GeneCorner. This plasmid was deposited by Dr M. Baens.|
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.