Last data update: 24 January 2024 16:39 CET
Plasmid name: pCMF2E-hFADD (LMBP 4585)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p4585.gb
(View with Genome Compiler) p4585.txt p4585.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human Fas receptor-associated death domain cDNA (FADD) Human FK506-binding protein prolyl isomerase 1A cDNA (FKBP1A, FKBP-12, FKBP12C, PKC12, PPIASE, GeneID 2280) Rous sarcoma virus (RSV) tyrosine kinase gene (v-src); myristoylation-targeting sequence, N-terminal Influenza HA epitope encoding the haemagglutinin tagging peptide; C-terminal |
| Promoter: | Human cytomegalovirus immediate early promoter (CMV-IE) and enhancer Phage T7 gene 10 promoter (T7g10) |
| Ribosome binding site: |
- |
| Terminator: | Rabbit β-globin polyadenylation signal (β-globin 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: | pCMF2E |
| Further information: | The plasmid was constructed by inserting a SpeI-XbaI fragment, containing the hFADD coding sequence, into the SpeI opened pCMF2E vector. The plasmid is designed for high-level expression of hFADD, fused to the N-terminal v-src myristoylation-targeting peptide, the N-terminal tandem repeat of hFKBP1A and the C-terminal HA epitope, under control of the strong hCMV-IE promoter and enhancer in mammalian cells. hFKBP1A is a member of the immunophilin protein family, which plays a role in immunoregulation and basic cellular processes involving protein folding and trafficking. This encoded protein is a cis-trans prolyl isomerase that binds the immunosuppressants FK506 and rapamycin. The fusion protein can be dimerized upon AP1510 (synthetic analogue of FK1012 ligand) administration. The v-src myristoylation-targeting sequence directs the fusion protein to cellular membranes. The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727114.1. The nucleotide sequence of the hFADD cDNA corresponds with the EMBL Nucleotide Sequence Database accession number U24231.1. |
| EMBL Accession number: | U24231.1, view at EMBL, GenBank, DDBJ LT727114.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 17/12/2003 |
| Sequence detail: | Nucleotide sequence of the fusion gene:
------- v-src myristoylation-targeting sequence ------>
5' ... GAATTCGCGCGT.ATG.GGG.AGT.AGC.AAG.AGC.AAG.CCT.AAG.GAC.CCC.AGC.CAG.CGC.TCT.AGA
EcoRI Met Gly Ser Ser Lys Ser Lys Pro Lys Asp Pro Ser Gln Arg Ser Arg
*** XbaI
SalI|XhoI
------------- hFKBP1A ------------> | ------------- hFKBP1A ------------>
GGA.GTG.CAG.GTG ... CTA.AAA.CTG.GAA.G|TC.GAG.GGC.GTG.CAG.GTG ... CTA.AAA.CTG.GAA
Gly Val Gln Val Leu Lys Leu Glu Val Glu Gly Val Gln Val Leu Lys Leu Glu
XbaI|SpeI
------------------ hFADD -----------------> |
1 208 | -------- HA epitope
ACT.AGT.ATG.GAC.CCG.TTC.CTG ... ACC.TCC.GAA.GCG.TCC.T|CT.AGT.TAT.CCG.TAC.GAC.GTA
Thr Ser Met Asp Pro Phe Leu Thr Ser Glu Ala Ser Ser Ser Tyr Pro Tyr Asp Val
SpeI ***
-------------->
CCA.GAC.TAC.GCA.TAA.GAAAAGTGAGGATCCTGAGAA ... 3'
Pro Asp Tyr Ala +++ BamHI
***: start codon.
+++: termination codon.
Punctuation indicates reading frame. |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BamHI, BstXI/PvuI, EcoRI/PstI, NcoI/SalI and SpeI/XbaI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Prof. Dr P. Vandenabeele(1) (2). It was constructed by G. Van Loo(1) (2). (1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium (2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | - |
| Related plasmid reference: | Kawahara et al., J. Cell Biol. 143 (1998), 1353-1360 [PMID: 9832562] Spencer et al., Curr. Biol. 6 (1996), 839-847 [PMID: 8805308] Vanden Berghe et al., J. Biol. Chem. 279 (2004), 7925-7933 [PMID: 14668343] |
| 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: |
pCMF2E-hFADD (LMBP 4585) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr P. Vandenabeele . |
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.