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GeneCorner plasmid details

Last data update: 26 November 2020 04:30 CET

Plasmid name: pCD-MK (LMBP 5536)

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Price category: Cat. 3 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p5536.gb (View with Genome Compiler)
p5536.txt
p5536.pdf
Sequence
analysis results
Genecorner:

Sanger: 144929.fasta

NGS: c03-gc-dec2018.fasta

Cloned DNA: Human MALT1 paracaspase cDNA (MALT1, MLT, PCASP1, paracaspase 1, GeneID 10892); with Kozak consensus sequence
Myc epitope; C-terminal
Histidine tag (His-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
Ribosome
binding site:
-
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: pcDNA3.1/myc-HisA
Further information: The plasmid is designed for high-level expression of hMLT, fused to the C-terminal Myc epitope and histidine tag, under control of the strong hCMV-IE promoter and enhancer in mammalian cells.
There is uncertainty about the presence of a second enhancer in the SV40 early promoter. The absence of a second enhancer in this promoter may cause bacterial leakage expression of the Tn5 neomycin resistance gene. This would cause transformed E. coli cells to be resistant to kanamycin, although growth should be reduced compared to growth on medium containing ampicillin.
The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT726920.1.
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-MLT.
EMBL Accession number: AF130356.2, view at EMBL, GenBank, DDBJ
LT726920.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 28/11/2007
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: Alw44I/HindIII, ApaI/BamHI, EcoRI/NcoI, NdeI, PauI/PstI and PdmI/SalI.

This plasmid has also 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 Dr M. Baens(1).
(1) Department of Human Genetics, KULeuven, Belgium
Plasmid reference: -
Related plasmid reference: Noels et al., J. Biol. Chem. 282 (2007), 10180-10189 [PMID: 17287209]
Elton et al., FEBS J. 283 (2016), 403-412 [PMID: 26573773] [DOI: 10.1111/febs.13597]
Staal et al., EMBO J. 30 (2011), 1742-1752 [PMID: 21448133] [DOI: 10.1038/emboj.2011.85]
Afonina et al., EMBO Rep. 17 (2016), 914-927 [PMID: 27113748] [DOI: 10.15252/embr.201642109]
Staal et al. (2016) [DOI: dx.doi.org/10.1101/046789]
Staal et al., bioRxiv (2016) [DOI: 10.1101/046789]
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 MC1061
Host reference: Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493]
Related host reference: Brigé et al., Biochem. J. 394 (2006), 335-344 [PMID: 16293111]
Cultivation medium: LB-Lennox + ampicillin (100 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
Cultivation remark: -
Other culture collection numbers: -

Refer in your Materials and Methods:

pCD-MK (LMBP 5536) 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.

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