Last data update: 24 January 2024 16:39 CET
Plasmid name: pEF-BOShFasRI (LMBP 3900)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p3900.gb
(View with Genome Compiler) p3900.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human TNF receptor superfamily, member 6 cDNA (FAS); intracellular region (codon 190-335) |
| Promoter: | Human elongation factor 1α promoter (EF1α) Escherichia coli lac operon promoter Simian virus 40 early promoter (SV40 early) |
| Ribosome binding site: |
- |
| Terminator: | Human granulocyte macrophage colony stimulating factor polyadenylation signal (GM-CSF polyA) |
| Selection marker: | Ampicillin (amp) |
| Replicon: | Bacteriophage M13 origin Escherichia coli plasmid pMB1 origin Simian virus 40 bidirectional origin (SV40) |
| Host range: | Escherichia coli Mammalian cells; SV40 permissive cells |
| Parental clone: | pEF-BOS; pEF-BOShFasR |
| Further information: | This plasmid was constructed as follows: the intracellular part of the human Fas antigen (from codon 190-335) from pEF-BOShFasR was amplified by PCR. The fragment was digested using EcoRI-NheI, and a 3-point ligation was carried out using the former fragment, the EcoRI-ScaI fragment (3233 bp) and the ScaI-XbaI fragment (1995 bp) of pEF-BOS. The correct sequence has been confirmed by sequencing. When cloning a fragment downstream from the lac promoter it may be advisable to use lacI(q) strains in order to prevent fortuitous expression of a possibly noxious polypeptide. The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727279.1. Other name of the plasmid is pEFhFasRic. |
| EMBL Accession number: | LT727279.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 28/06/1999 |
| Sequence detail: | Linker 1: 5' GCCGAATTCCACC.ATG.GTG.AAG.AGA.AAG.GAA.GTA.CAG.AAA 3'
EcoRI
Linker 2: 5' GCCGCT.AGC.GAC.CAA.GCT.TTG.GAT.TTC.ATT.TCT.G 3'
NheI
Nucleotide sequence at the start of the intracellular part of human FasR:
------------------ hFasRI ----------------->
190 191
5' ... GTGAGGAATTCCACC.ATG.GTG.AAG.AGA.AAG.GAA.GTA.CAG.AAA.ACA.TGC.AGA ... 3'
EcoRI *** Val Lys Arg Lys Glu Val Gln Lys Thr Cys Arg
NcoI
Nucleotide sequence at the end of the intracellular part of human FasR:
------ hFasRI ----->
335 -------- 3'UTR of hGM-CSF -------->
5' ... ATC.CAA.AGC.TTG.GTC.GCT.AGA.GTG.AGG.GTC.CCC.ACC.TGG.GAC.CCT.TGA ... 3'
Ile Gln Ser Leu Val Ala Arg Val Arg Val Pro Thr Trp Asp Pro +++
HindIII NheI/XbaI fusion
***: Start codon.
+++: Termination codon.
Punctuation indicates reading frame. |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: EcoRI, HindII, HindIII and XhoI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Dr D. Vercammen(1) (2) and Prof. Dr P. Vandenabeele (1) (2). (1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium (2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | - |
| 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 |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pEF-BOShFasRI (LMBP 3900) is available at BCCM/GeneCorner. This plasmid was deposited by Dr D. Vercammen and 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.