Last data update: 24 January 2024 16:39 CET
Plasmid name: pCDNA3-F-HLH-NvPCASPt1B_intron (LMBP 9801)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | not available |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Nematostella vectensis type 1 paracaspase cDNA (PCASP-t1B, MALT1 homolog); intron HLH dimerization domain; N-terminal FLAG epitope tag; N-terminal |
| Promoter: | Human cytomegalovirus immediate early promoter (CMV-IE) and enhancer Simian virus 40 early promoter (SV40 early) Phage T7 gene 10 promoter (T7g10) 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-F-HLH |
| Further information: | The plasmid was constructed by PCR amplifying the N. vectensis PCASP-t1B coding sequence from a cDNA library and cloning it into the BamHI/ApaI-opened pCDNA3-F-HLH vector. Type 1 paracaspases originated before the last common ancestor of cnidarians (e.g. Nematostella) and bilaterans. Cnidarians have 2 paralog type 1 paracaspases and a homolog of Bcl10. Intriguingly, different bilateran lineages seem to have 2 different type 1 paracaspases, which is correlated with the presence or absence of Bcl10. Vertebrates, mollusks and acorn worm have Bcl10 and the paracaspases are similar. Nematodes, Insects, Annelids do not have Bcl10 and have a less similar paracaspase. It was already reported that the "A" paralog from Nematostella lacks MALT1-like activity (Hulpiau et al 2015). The N. vectensis PCASP-t1B coding sequence in this plasmid still contains a 162 bp intron sequence close to the N-terminus, which causes a very short expressed fragment. For future functional studies, this intron needs to be deleted. The nucleotide sequence of the N. vectensis PCASP-t1B coding sequence corresponds with the Genbank accession number XM_001629469.1. |
| EMBL Accession number: | XM_001629469.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 03/09/2019 |
| Sequence detail: | Primers used to amplify the N. vectensis PCASP-t1B coding sequence:
Forward: 5' CCATGGATCC.ATG.GCC.GAC.AGC.TTG.TGT.ATT.A
BamHI ***
Reverse: 5' CTTCGGGCCC.ACT.GGG.CTG.CAC.TAT.ATG
ApaI
***: start codon
Punctuation indicates reading frame. |
| 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 Prof. Dr R. Beyaert(1) (2) and Dr J. Staal(1) (2). (1) Center for Inflammation Research, VIB, Ghent, Belgium (2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | - |
| Related 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-F-HLH-NvPCASPt1B_intron (LMBP 9801) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Beyaert and Dr J. Staal . |
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.