Last data update: 24 January 2024 16:39 CET
Plasmid name: pGEXGSTp65(12-317) (LMBP 5055)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p5055.gb
(View with Genome Compiler) p5055.txt p5055.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Schistosoma japonicum Sj26 antigen glutathione S-transferase gene (GST) Thrombin recognition site; N-terminal Human nuclear factor κB cDNA (NF-κB); fragment of the p65 (RELA) subunit Escherichia coli lac repressor gene (lacI) Escherichia coli lac Z gene (lacZ); fragment, incl. α part |
| Promoter: | Escherichia coli hybrid tryptophan/lacUV5 promoter (tac) Escherichia coli lac repressor promoter; mutant (lacI(q)) Escherichia coli lac operon promoter |
| Ribosome binding site: |
Ribosome binding site (RBS) of the Escherichia coli lac Z gene (lacZ) |
| Terminator: | - |
| Selection marker: | Ampicillin (amp) |
| Replicon: | Escherichia coli plasmid pMB1 origin |
| Host range: | Escherichia coli |
| Parental clone: | pGEX-2T |
| Further information: | The plasmid was constructed by inserting a 924 bp BamHI fragment, containing the human NF-κB p65 cDNA fragment encoding the amino acids 12 up to and including 317, into the BamHI site of pGEX-2T. As a result, the p65(12-317) coding sequence was fused in phase to the N-terminal coding sequence of the S. japonicum GST gene and the N-terminal thrombin recognition sequence. Transcription of the GST-p65(12-317) fusion gene is controlled by the binding of the lac repressor, encoded by the lacI gene, to the lac operator of the E. coli tac promoter. pGEXGSTp65(12-317) can be used for the synthesis of the GST-p65(12-317) fusion protein in E. coli. The GST carrier can be cleaved from the fusion protein by digestion with the site-specific protease thrombin. The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727075.1. The nucleotide sequence of the pGEX-2T vector corresponds with the EMBL Nucleotide Sequence Database accession number U13850.1. The nucleotide sequence of the human NF-κB p65 coding sequence corresponds with the EMBL Nucleotide Sequence Database accession number M62399.1. Name mentioned in Delerive et al. (1999) is pGexp65 12-317. Name mentioned in Vermeulen et al. (2003) is pGEXp65 12-317. Other name of the plasmid is pGEX-p65dC. |
| EMBL Accession number: | U13850.1, view at EMBL, GenBank, DDBJ M62399.1, view at EMBL, GenBank, DDBJ LT727075.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 27/07/2005 |
| Sequence detail: | Nucleotide sequence of the fusion gene:
----------- tac promoter ---------->------------- lacZ RBS ---------
5'...GTTGACAATTAATCATCGGCTCGTATAATGTGTGGAATTGTGAGCGGATAACAATTTCACACAGGAA
HindII
---> ----------------------- GST ---------------------->
ACAGTATTC.ATG.TCC.CCT.ATA.CTA.GGT ... GGC.GAC.CAT.CCT.CCA.AAA.TCG.GAT
Met Ser Pro Ile Leu Gly Gly Asp His Pro Pro Lys Ser Asp
***
-- thrombin recogn. --> ------------------------------ p65f ------------
12 276
CTG.GTT.CCG.CGT.GGA.TCC.GAG.CCA.GCC.CAG.GCC.TCT...CGG.CCT.TCC.GAC.CGG...
Leu Val Pro Arg Gly Ser Glu Pro Ala Gln Ala Ser Arg Pro Ser Asp Arg
^BamHI StuI
---------------------->
317
ATC.ATG.AAG.AAG.AGT.CCT.GGA.TCC.CCG.GGA.ATT.CAT.CGT.GAC.TGA.CTGAC...3'
Ile Met Lys Lys Ser Pro Gly Ser Pro Gly Ile His Arg Asp +++
BamHI AvaI EcoRI
^: thrombin cleavage site.
***: start codon.
+++: termination codon.
Punctuation indicates reading frame. |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BamHI, BglI/HindII, EcoRV, PvuI and StuI/XmnI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Prof. Dr R.T. Hay(1). (1) Centre for Biomolecular Sciences, University of St. Andrews, St. Andrews, UK |
| Plasmid reference: | - |
| Related plasmid reference: | Vermeulen et al., EMBO J. 22 (2003), 1313-1324 [PMID: 12628924] Jaffray et al., Mol. Cell. Biol. 15 (1995), 2166-2172 [PMID: 7891711] Delerive et al., J. Biol. Chem. 274 (1999), 32048-32054 [PMID: 10542237] |
| 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: |
pGEXGSTp65(12-317) (LMBP 5055) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R.T. Hay. |
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.