Last data update: 24 January 2024 16:39 CET
Plasmid name: pCD-A7M8K-p20mut (LMBP 5538)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p5538.gb
(View with Genome Compiler) p5538.txt p5538.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human apoptosis inhibitor 2 cDNA (API2, BIRC3); N-terminal fragment (exon 1 --> 7) with Kozak consensus sequence Human MALT1 paracaspase cDNA (MALT1, MLT, PCASP1, paracaspase 1); mutated C-terminal fragment (exon 8 --> end) 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: | pCD-A7M8K |
| Further information: | The plasmid contains the N-terminal fragment of the hAPI2 coding sequence (exon 1 --> exon 7) fused to the mutated C-terminal fragment of the hMLT coding sequence (exon 8 --> end) and to the C-terminal Myc epitope and histidine tag. As compared to the wt hMLT cDNA, the predicted catalytic cysteine (C) codon at position 464, in the caspase(p20)-like active site of hMLT, was replaced by an alanine (A) codon, creating an extra PauI site. The plasmid is designed for high-level expression of the fusion protein under control of the strong hCMV-IE promoter and enhancer in mammalian cells. 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 LT726922.1. The nucleotide sequence of the wt hAPI2 cDNA corresponds with the EMBL Nucleotide Sequence Database accession number BC037420.1. 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-AMK p20mut. |
| EMBL Accession number: | BC037420.1, view at EMBL, GenBank, DDBJ AF130356.2, view at EMBL, GenBank, DDBJ LT726922.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 28/11/2007 |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: Alw44I/HindIII, ApaI/BamHI, NcoI, NdeI, PauI/PstI and PdmI/SalI. |
| 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: | Staal et al., EMBO J. 30 (2011), 1742-1752 [PMID: 21448133] [DOI: 10.1038/emboj.2011.85] Coornaert et al., Nat. Immunol. 9 (2008), 263-271 [PMID: 18223652] [DOI: 10.1038/ni1561] 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 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-A7M8K-p20mut (LMBP 5538) 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.