Last data update: 24 January 2024 16:39 CET
Plasmid name: pGal4-p65f1-285 (LMBP 4713)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p4713.gb
(View with Genome Compiler) p4713.txt p4713.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 1593 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 1 up to and including 286. 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 LT727172.1. The nucleotide sequence of pGal4-p65 was compiled in accordance with the description in Schmitz et al. (1991). The fusion region between GALbd and the hNF-κB p65 subunit in pGal4-p65 was sequenced (F. Molemans, Dept of Biomedical Molecular Biology, Ghent University, Belgium). The nucleotide sequence of the hNF-κB p65 coding sequence corresponds with the EMBL Nucleotide Sequence Database accession number M62399.1. 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 1_285. Other names of the plasmid are pABgal4-p65(1-285) and pABgal4p65deltaC. |
| EMBL Accession number: | M62399.1, view at EMBL, GenBank, DDBJ LT727172.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 03/10/2003 |
| 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 -------------->
147 ------------------- linker ----------------
5' ... AAA.GGT.CAA.AGA.CAG.TTG.ACT.GTA.TCG.CCG.GAA.TTC.CCG.GGT.GTC.GAC.CAG.GGG.ACC.CCG
Lys Gly Gln Arg Gln Leu Thr Val Ser Pro Glu Phe Pro Gly Val Asp Gln Gly Thr Pro
HindII EcoRI AvaI HindII
SmaI SalI
--------------- p65f ----------------->
--> 1
GCC.ATG.GAC.GAA.CTG.TTC.CCC.CTC.ATC.TTC ... 3'
Ala Met Asp Glu Leu Phe Pro Leu Ile Phe
***
NcoI
StyI
Nucleotide sequence at the end of the hNF-kB p65 subunit cDNA fragment:
---------- p65f ------> --- SV40 polyA early --->
286
5' ... GAG.CCC.ATG.GAA.TTC.GAT.ATC.AAG.CTT.GAT.CCG.TGG.CGG.AAT.TAA.CTTGTTTATTGCAGCTTA ... 3'
Glu Pro Met Glu Phe Asp Ile Lys Leu Asp Pro Trp Arg Asn +++
NcoI EcoRI EcoRV HindIII
StyI
***: start codon.
+++: termination codon.
Punctuation indicates reading frame. |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: AvaI/HaeII, BamHI/BglII, EcoRI, HindIII, KpnI/SmaI, PstI and 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-p65f1-285 (LMBP 4713) 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.