Last data update: 24 January 2024 16:39 CET
Plasmid name: pcDNA3-FLAG-hIRAK1-KD (LMBP 5626)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p5626.gb
(View with Genome Compiler) p5626.txt p5626.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human interleukin-1 receptor-associated kinase 1 cDNA (IRAK1); mutated sequence FLAG epitope tag; N-terminal |
| Promoter: | Human cytomegalovirus immediate early promoter (CMV-IE) and enhancer Phage SP6 promoter Phage T7 gene 10 promoter (T7g10) Simian virus 40 early promoter (SV40 early) 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 |
| Further information: | The plasmid was created by ligating the human IRAK1 cDNA into a FLAG-tagged pcDNA3 vector. The human IRAK1-KD mutant was generated by site-directed mutagenesis, converting the aspartic acid codon (D) at position 340 into an asparagine codon (N). Assessment of the kinase activity of insect cells expressing human IRAK1-KD confirmed that the mutant was unable to auto-phosphorylate compared to insect cells expressing wild type human IRAK1. The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT726933.1 The nucleotide sequence of the wild type human IRAK1 coding sequence corresponds with the EMBL Nucleotide Sequence Database accession number L76191.1. |
| EMBL Accession number: | L76191.1, view at EMBL, GenBank, DDBJ LT726933.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 06/02/2009 |
| Sequence detail: | Nucleotide sequence at the start of the human IRAK1 fragment:
|--> FLAG tag -->| |--> hIRAK1
5' ... ATG.GAT.TAC.AAA.GAC.GAT.GAC.GAT.AAA.GAA.TTC.ACC.ATG.GCC. ... 3'
*** ***
***: start codon.
Punctuation indicates reading frame. |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: HindIII/XbaI, NcoI, PvuI and XmnI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Prof. Dr J. Tschopp(1). (1) Department of Biochemistry, University of Lausanne, Switzerland |
| Plasmid reference: | - |
| Related plasmid reference: | Volpe et al., FEBS Lett. 419 (1997), 41-44 [PMID: 9426216] |
| 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 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pcDNA3-FLAG-hIRAK1-KD (LMBP 5626) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr J. Tschopp. |
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.