Last data update: 03 December 2020 04:26 CET
Plasmid name: pCAGGS-E-hABIN1-C (LMBP 5129)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner non-core plasmid|
|Cloned DNA:||Human A20-binding inhibitor of NF-κB activation 1 cDNA (ABIN-1, TNIP1, NAF1); C-terminal fragment
|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
|Further information:||The plasmid was constructed by inserting a 794 bp NotI-XhoI PCR fragment, containing the codons 376 up to and including 636 of the human A20-binding inhibitor of NF-κB activation 1 (hABIN-1) coding sequence, into the NotI-XhoI opened pCAGGS-E-hABIN1-P299A vector.
pCAGGS-E-hABIN1-C is useful for highly efficient expression of the C-terminal fragment of hABIN-1, fused in phase to the N-terminal E-tag, under the control of the chicken β-actin/rabbit β-globin hybrid promoter (AG) and the human CMV-IE enhancer in various mammalian cells.
The AG promoter sequence consists of the chicken β-actin promoter, the first exon and part of the first intron (that seems to have a strong enhancer-like activity) linked to a rabbit β-globin fragment, consisting of a 3' part of the second intron (inclusive a branch point which is required for normal splicing reactions) and a 5' part of the third exon.
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 hABIN-1 cDNA was obtained from Genbank (Accession number NM_006058.2).
|EMBL Accession number:||NM_006058.2, view at GenBank|
|Latest sequence update:||21/12/2005|
Primers used to amplify the C-terminal fragment of the hABIN-1 coding sequence: 376 380 forward: 5' ... CGGGATCCTAGCGGCCGCC.ATG.GAG.GAG.ACC.GAC.AAG.G ... 3' BamHI NotI *** 636 635 reverse: 5' ... GCCGCTCGAG.TCA.CTG.AGG.CCC.CTC.ACG ... 3' XhoI +++ ***: start codon. +++: termination codon. Punctuation indicates reading frame.
|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
|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-hABIN1-C (LMBP 5129) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Beyaert .|
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.