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GeneCorner plasmid details

Last data update: 21 October 2020 04:26 CEST

Plasmid name: pAS2-IKKgamma-MUT2 (LMBP 5500)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: not available
Sequence
analysis results
Genecorner:

-

Cloned DNA: Human NF-κB inhibitor kinase γ cDNA (IKKγ, NEMO, FIP3, IKBKG); mutated ABIN Homology Domain 2 (AHD2)
Saccharomyces cerevisiae GAL4 DNA-binding domain (GALbd)
Influenza HA epitope encoding the haemagglutinin tagging peptide; N-terminal
Promoter: Saccharomyces cerevisiae alcohol dehydrogenase promoter (ADH1)
Escherichia coli lac operon promoter
Ribosome
binding site:
-
Terminator: Saccharomyces cerevisiae alcohol dehydrogenase terminator (ADH1)
Saccharomyces cerevisiae iso-1-cytochrome C terminator (CYC1)
Saccharomyces cerevisiae 2 micron plasmid (2μ) FLP terminator
Selection marker: Ampicillin (amp)
Cycloheximide (CYH2)
TRP1; auxotrophic
Replicon: Escherichia coli plasmid pMB1 origin
Phage f1 origin
Saccharomyces cerevisiae 2 micron plasmid origin (2μ); incl. region conferring stability (STB)
Host range: Escherichia coli
Saccharomyces cerevisiae; trp1(-)
Parental clone: pM-IKKgamma-MUT2; pAS2
Further information: The plasmid was constructed by inserting an EcoRI(filled-in)/SalI fragment, containing the mutated human IKKγ sequence, into the MscI-SalI opened pAS2 vector. As a result, the inserted DNA is fused in phase to the N-terminal HA epitope and the S. cerevisiae GALbd domain.
Yeast two hybrid
The plasmid contains full length IKKγ with mutation in AHD2 (DF -> NA).
Other name of the plasmid is pAS2-IKKγ-MUT2.
EMBL Accession number: AJ271718, view at EMBL, GenBank, DDBJ
Latest sequence update: 03/03/2008
Sequence detail:
Amino acid sequece of human IKKgamma:

IKKgamma wild type: 300 VLKAQADIYKADFQAERQAREKL 322

IKKgamma MUT1     : 300 VLKEEADIYKADFQAERQAREKL 322
                           ** 

IKKgamma MUT2     : 300 VLKAQADIYKANAQAERQAREKL 322
                                   **

IKKgamma MUT3     : 300 VLKAQADIYKADFQAERQAAQKL 322
                                           **
*: mutated amino acid
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 K. Heyninck(1) (2) and 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
Plasmid reference: -
Related plasmid reference: Heyninck et al., FEBS Lett. 536 (2003), 135-140 [PMID: 12586352]
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)
Cultivation temperature: 37°C
Biosafety level: L1
Other culture collection numbers: -

Refer in your Materials and Methods:

pAS2-IKKgamma-MUT2 (LMBP 5500) is available at BCCM/GeneCorner. This plasmid was deposited by Dr K. Heyninck and 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.

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