Last data update: 24 January 2024 16:39 CET
Plasmid name: pcDNA3.1-F-HLH-TaPCASP-HA (LMBP 9574)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p9574.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Trichoplax adherens type 2 paracaspase cDNA (type 2 PCASP) FLAG epitope tag; N-terminal HLH dimerization domain; N-terminal Influenza HA epitope encoding the haemagglutinin tagging peptide; 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-F-HLH-Ig2-VSV-MALT1C-EARmut-HA; pcDNA3.1-Flag-TaPCASP |
| Further information: | The plasmid was created by replacing the Ig2-VSV-MALT1C-EARmut cassette of pcDNA3.1-F-HLH-Ig2-VSV-MALT1C-EARmut-HA with the T. adherens PCASP coding sequence obtained from pcDNA3.1-Flag-TaPCASP as a BamHI/EcoRI fragment. Vertebrates typically contain 3 paracaspase paralogs, but all but 1 (PCASP1, MALT1) have been lost in mammals. A direct relative to the ancestral PCASP3 can be found as far back as sea urchins. Among other invertebrates, there are many different lineages of paracaspases. Advanced invertebrates have the "modern" (MALT1-like) DD-Ig1-Ig2-PCASP-Ig3 domain architecture of type 1 paracaspases, whereas primitive invertebrates (sponges, trichoplax,...) have a PCASP-only type protein, classing them as type 2 paracaspases. Here the T. adherens type 2 PCASP has been cloned behind the activating ETV6 HLH domain in order to study functional conservation and redundancy between the paracaspases. Other name of the plasmid is pCDNA3-F-HLH-TaPCASP-HA. |
| EMBL Accession number: | - |
| Latest sequence update: | 01/02/2016 |
| Authenticity test: | The plasmid still needs to be subjected to the authenticity test. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| 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 |
| Plasmid reference: | Hulpiau et al., Cell. Mol. Life Sci. 73 (2016), 1103-1116 [PMID: 26377317] [DOI: 10.1007/s00018-015-2041-9] |
| 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: | Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660] Rodriguez-Quinones et al., Focus 15 (1993), 110-112 Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791] [DOI: 10.1016/s0022-2836(83)80284-8] Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187] Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051] |
| Cultivation medium: | LB-Lennox + ampicillin (100 μg/ml) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pcDNA3.1-F-HLH-TaPCASP-HA (LMBP 9574) is available at BCCM/GeneCorner. This plasmid was deposited by Dr J. Staal and Prof. Dr R. Beyaert and was published in Hulpiau et al., 2016. |
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.