Last data update: 24 January 2024 16:39 CET
Plasmid name: pGal4-p65f286-550 (LMBP 4709)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p4709.gb
(View with Genome Compiler) p4709.txt p4709.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Saccharomyces cerevisiae GAL4 DNA-binding domain (GALbd) Human nuclear factor κB cDNA (NF-κB); fragment of the p65 (RELA) subunit |
| Promoter: | Rous sarcoma virus long terminal repeat (RSV-LTR) Simian virus 40 early promoter (SV40 early) |
| Ribosome binding site: |
- |
| Terminator: | 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 |
| Parental clone: | pGal4-p65 |
| Further information: | This plasmid was derived from pGal4-p65 by removing the 885 bp EcoRI fragment. The plasmid contains the S. cerevisiae GALbd domain, composed of the first 147 codons from GAL4, fused to the hNF-κB p65 cDNA fragment encoding the amino acids 285 up to and including 551. The GALbd-p65f fusion protein can be used as a bait protein in two-hybrid screening. The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727168.1. The nucleotide sequence of pGal4-p65 was compiled in accordance with the description in Schmitz et al. (1991). The nucleotide sequence of the hNF-κB p65 coding sequence corresponds with the EMBL Nucleotide Sequence Database accession number M62399 (verison SV 1). The nucleotide sequence of the 3' UTR of the hNF-κB p65 subunit was retrieved from the pRc/CMV-p65 sequence file provided by S. Plaisance (Dept of Biomedical Molecular Biology, Ghent University, Belgium). The nucleotide sequence upstream of GALbd in pGal4-p65 (from nucleotide position 3841 up to and including 4920) was analysed by BCCM/GeneCorner. Name mentioned in Schmitz et al. (1991) is GAL4-p65 286_550. Other names of the plasmid are pABgal4-p65(286-551) and pABgal4p65deltaN. |
| EMBL Accession number: | M62399, view at EMBL, GenBank, DDBJ LT727168.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 25/09/2003 |
| Sequence detail: | Nucleotide sequence at the start of the S. cerevisiae GAL4 DNA-binding domain:
---------- GALbd ------------->
2
5' ... ACTG.ATG.GAC.TCC.CAG.CAG.CCA.GAT.CTG.AAG.CTA.CTG.TCT.TCT.ATC.GAA ... 3'
Met Asp Ser Gln Gln Pro Asp Leu Lys Leu Leu Ser Ser Ile Glu
*** BglII
Nucleotide sequence at the GALbd-p65f fusion:
----------------- GALbd --------------> ----------------- p65f ------------------->
147 285
5' ... AAA.GGT.CAA.AGA.CAG.TTG.ACT.GTA.TCG.CCG.GAA.TTC.CAG.TAC.CTG.CCA.GAT.ACA.GAC.GAT ... 3'
Lys Gly Gln Arg Gln Leu Thr Val Ser Pro Glu Phe Gln Tyr Leu Pro Asp Thr Asp Asp
HindII EcoRI PvuI
Nucleotide sequence at the end of the hNF-kB p65 subunit cDNA fragment:
-------------------------------- p65f ---------------------------->
551
5' ... ATT.GCG.GAC.ATG.GAC.TTC.TCA.GCC.CTG.CTG.AGT.CAG.ATC.AGC.TCC.TAA.GGGGGTGACGCCTGC ... 3'
Ile Ala Asp Met Asp Phe Ser Ala Leu Leu Ser Gln Ile Ser Ser +++
***: start codon.
+++: termination codon.
Punctuation indicates reading frame. |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BamHI, EcoRI, KpnI/SmaI and PvuI/XhoI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Dr M.L. Schmitz(1). (1) Department of Chemistry and Biochemistry, University of Bern, Switzerland |
| Plasmid reference: | Schmitz et al., EMBO J. 10 (1991), 3805-3817 [PMID: 1935902] |
| 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: |
pGal4-p65f286-550 (LMBP 4709) is available at BCCM/GeneCorner. This plasmid was deposited by Dr M.L. Schmitz and was published in Schmitz et al., 1991. |
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.