Last data update: 20 January 2021 04:26 CET
Plasmid name: pCAGGS-E-mABIN-1(388-560) (LMBP 5365)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner non-core plasmid|
|Depositor's sequence:||not available|
Mouse A20-binding inhibitor of NF-κB activation 1 cDNA (ABIN-1, TNIP1); C-terminal fragment encoding amino acids (388-560)|
|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; pCAGGSEhA20|
|Further information:||The plasmid was constructed as follows: a splice variant (388-560) of mouse ABIN-1 was generated by PCR, the PCR fragment was NotI-SmaI digested and cloned in a 3-point ligation with a NotI-PvuI fragment of pCAGGS-E-A20 and a MscI-PvuI fragment of pCAGGS.
pCAGGS-E-mABIN-1(388-560) is useful for highly efficient expression of a C-terminal fragment (aa 388-560) of mouse A20-binding inhibitor of NF-kappa B activation (ABIN1), under the control of the chicken β-actin/rabbit β-globin hybrid promoter (AG) 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 mABIN-1 cDNA corresponds with the EMBL Nucleotide Sequence Database accession number AJ242778.1.
|EMBL Accession number:||AJ242778.1, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||16/05/2007|
The primers used in PCR to amplify mABIN-1(10) cDNA were: forward: 5' ATAAGAATGCGGCCGCAGAGATGGAAGAGACCG 3' NotI reverse: 5' TCCCCCGGGTCAGGCGCCGCAGACGTGCTCAGGG 3' SmaI
|Authenticity test:||The plasmid still needs to be subjected to the authenticity test.|
|History of deposit:||This plasmid was deposited by Prof. Dr R. Beyaert(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
|Related plasmid reference:||Heyninck et al., FEBS Lett. 442 (1999), 147-150 [PMID: 9928991]
|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-mABIN-1(388-560) (LMBP 5365) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Beyaert .|