Last data update: 24 January 2024 16:39 CET
Plasmid name: pEFCAThFasRS (LMBP 3901)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | not available |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Chloramphenicol acetyltransferase gene (cat) Human Fas antigen; intracellular region Strep-tag |
| Promoter: | Human elongation factor 1α promoter (EF1α) Escherichia coli lac operon promoter Simian virus 40 early promoter (SV40 early) |
| Ribosome binding site: |
Ribosome binding site (RBS) of the Escherichia coli lac Z gene (lacZ) |
| 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-BOSICES-T; pCDM8CAT; pCDM8hFasR |
| Further information: | This plasmid was constructed as follows: CAT and the intracellular region of human Fas were amplified by PCR. These reactions add a linker (L), consisting of 3 x (GGSG), downstream of CAT and upstream of intracellular Fas. CAT-L and L-Fas were digested using KpnI-BamHI and BamHI-NheI, respectively, and a 3-point ligation was carried out using the former fragments and KpnI-NheI opened pEF-BOSICES-T. 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. |
| EMBL Accession number: | - |
| Latest sequence update: | |
| Sequence detail: | Nucleotide sequence of PCR fragments: 5' GGCGGTACCACCATGGAGAAAAAAATCACTGGATATA 3' 5' GCCGCTACCGCCCCCGGATCCCCCTCCGGAACCTCCCGCCCCGCCCTGCCACTCATCGCAG 3' Kozak sequence: 5' CCACCATGG 3' Nucleotide sequence of PCR fragments: 5' GGAGGTTCCGGAGGGGGATCCGGGGGCGGTAGCGGCAAGAGAAAGGAAGTACAGAAAACAT 3' 5' GCCGCTAGCGACCAAGCTTTGGATTTCATTTCTG 3' |
| Authenticity test: | The plasmid still needs to be subjected to the authenticity test. |
| 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 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pEFCAThFasRS (LMBP 3901) 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.