Last data update: 29 October 2020 04:15 CET
Plasmid name: pCAGGS-E-hPELI1-DR (LMBP 5191)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
|Cloned DNA:||Human pellino 1 cDNA (PELI1); N-terminal part encoding amino acids 1-332, RING-domain deleted (PELI1-DR)
|Promoter:||Chicken β-actin/rabbit β-globin hybrid promoter (AG)
Human cytomegalovirus immediate early promoter (CMV-IE); enhancer only
Escherichia coli lac operon promoter
|Terminator:||Rabbit β-globin polyadenylation signal (β-globin polyA)
Simian virus 40 polyadenylation signal (SV40 polyA)
|Selection marker:||Ampicillin (amp)|
|Replicon:||Escherichia coli plasmid pMB1 origin
Simian virus 40 bidirectional origin (SV40)
|Host range:||Escherichia coli
Mammalian cells; SV40 permissive cells
|Parental clone:||pCAGGS-E-hPELI1; pCAGGS|
|Further information:||The plasmid was constructed as follows: 1) a C-terminal deletion mutant of the human PELI1 cDNA, encoding amino acids 1 up to 332 and lacking most of the RING motif, was subcloned from pCAGGS-E-hPELI1 via PCR; 2) the NotI/XhoI digested PCR amplicon was fused in frame to the E-tag of a NotI/XhoI opened pCAGGS-based expression plasmid.
pCAGGS-E-hPELI1-DR is useful for highly efficient expression of E-tagged hPELI1-DR under the control of the AG promoter and the human CMV-IE enhancer in various mammalian cells.
When cloning a fragment downstream from the lac promoter it may be advisable to use lacI(q) strains in order to prevent fortuitous expression of a possibly noxious polypeptide.
The nucleotide sequence of the human PELI1-DR cDNA corresponds with the EMBL Nucleotide Sequence Database accession number AF302505.1, except for the nucleotides at positions 72, 149 and 243 of the coding sequence which do not change the amino acid sequence, and at position 149 which results in a N97T change compared to the EMBL record. The hPELI1-DR insert was sequenced by the depositor.
Other name of the plasmid is pCAGGS-E-hPeli-1DR.
|EMBL Accession number:||AF302505.1, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||16/04/2009|
The primers used in PCR to amplify hPELI1-DR cDNA were: forward : 5' ATAAGAATGCGGCCGCA.ATG.TTT.TCT.CCT.GAT.CAA.GAA.A 3' NotI *** reverse : 5' CCGCTCGAG.TTA.TTC.ACG.ATC.TTT.TCC.ATC.ACG.TTC 3' XhoI +++ ***: start codon. +++: termination codon.
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: Alw44I/SacII, HincII, NcoI/XmnI, NotI/XhoI and SspI.|
|History of deposit:||This plasmid was deposited by Prof. Dr R. Beyaert(1) (2). It was constructed by Dr R. Schauvliege(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
|Plasmid reference:||Schauvliege et al., FEBS Lett. 580 (2006), 4697-4702 [PMID: 16884718]
|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)|
|Other culture collection numbers:||-|
Refer in your Materials and Methods:
|pCAGGS-E-hPELI1-DR (LMBP 5191) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Beyaert and was published in Schauvliege et al., 2006.|
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.