Last data update: 24 January 2024 16:39 CET
Plasmid name: pFLAG-p65-S276A (LMBP 6089)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p6089.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human nuclear factor κB cDNA (NF-κB); mutated p65 (RELA) subunit FLAG epitope tag; N-terminal |
| Promoter: | Human cytomegalovirus immediate early promoter (CMV-IE) and enhancer Simian virus 40 early promoter (SV40 early) Phage T7 gene 10 promoter (T7g10) Escherichia coli lac operon promoter |
| Ribosome binding site: |
- |
| Terminator: | Human growth hormone 1 polyadenylation signal (GH1 polyA) |
| Selection marker: | Ampicillin (amp) |
| 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: | pFLAG-CMV-2; CMV-p65 |
| Further information: | The plasmid was created by introducing a S276A mutation in the human NF-κB p65 coding sequence of CMV-p65, using site-directed mutagenesis, after which the CDS was PCR amplified and cloned as a HindIII-EcoRV fragment into the HindIII-EcoRV opened pFLAG-CMV-2 vector. The nucleotide sequence of the wild type human NF-κB p65 coding sequence corresponds with the EMBL Nucleotide Sequence Database accession number M62399.1. Name mentioned in Wang and Baldwin (1998) is F-276A. Other name of the plasmid is pFLAG p65 S276A. |
| EMBL Accession number: | M62399.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 26/02/2009 |
| Sequence detail: | Mutagenesis primers resulting in S276A mutation:
Forward primer: 5' GCG.GCG.GCC.TGC.CGA.CCG.GGA.G 3'
^^ ^
Reverse primer: 5' CTC.CCG.GTC.GGC.AGG.CCG.CCG.C 3'
^^^
PCR primers used to amplify p65 cds:
Forward primer: 5' GAC.AAG.CTT.GAC.GAA.CTG.TTC.CCC.CTC.AT 3'
HindIII
Reverse primer: 5' GTCGATATC.TTA.GGA.GCT.GAT.CTG.ACT.C 3'
EcoRV +++
^: mutated nucleotide
+++: termination codon
Punctuation indicates reading frame. |
| 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 Prof. Dr A. Baldwin(1). (1) Lineberger Cancer Center, University of North Carolina at Chapel Hill, North Carolina, USA |
| Plasmid reference: | Wang et al., J. Biol. Chem. 273 (1998), 29411-29416 [PMID: 9792644] |
| Related plasmid reference: | Spooren et al., J. Biomed. Biotechnol. 2010 (2010), 275892 [PMID: 20204068] [DOI: 10.1155/2010/275892] |
| 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: |
pFLAG-p65-S276A (LMBP 6089) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr A. Baldwin and was published in Wang et al., 1998. |
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.