Last data update: 24 January 2024 16:39 CET
Plasmid name: pSV-Sport-Raf-1E (LMBP 5264)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p5264.gb
(View with Genome Compiler) p5264.txt p5264.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human v-raf-1 murine leukemia viral oncogene homolog 1 cDNA (hRaf1) E-tag; C-terminal |
| Promoter: | Phage SP6 promoter Phage T7 gene 10 promoter (T7g10) Escherichia coli lac operon promoter; mutant (lacUV5) 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: | pSV-Sport-Raf-1; pSV-Sport-Etag |
| Further information: | This plasmid was constructed as follows: 1) hRaf1 cDNA was amplified by PCR; 2) the amplicon was EcoRI digested and cloned into pSV-Sport1; 3) a NotI restriction site was introduced at the hRaf1 termination codon by site-directed mutagenesis; 4) the hRaf1 cDNA was finally inserted as an EcoRI-NotI fragment in the pSV-Sport-Etag expression vector, in frame with the C-terminal E-tag. 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 LT726992.1. The nucleotide sequence of the hRaf1 cDNA was obtained from Genbank (Accession number NM_002880.2). Other name of the plasmid is pSVSport h cRAF1E. |
| EMBL Accession number: | NM_002880.2, view at GenBank LT726992.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 07/02/2007 |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: AvaII, EcoRI/NotI, HaeII and XbaI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Y. Bruynooghe(1) (2) and Prof. Dr R. Beyaert(1) (2). (1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium (2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | Cornelis et al., Oncogene 24 (2005), 1552-1562 [PMID: 15674327] |
| Related plasmid reference: | Tran et al., J. Cell Sci. 130 (2017), 2371-2381 [PMID: 28576973] [DOI: 10.1242/jcs.186411] Cornelis et al., Mol. Cell 5 (2000), 597-605 [PMID: 10882096] |
| 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: |
pSV-Sport-Raf-1E (LMBP 5264) is available at BCCM/GeneCorner. This plasmid was deposited by Y. Bruynooghe and Prof. Dr R. Beyaert and was published in Cornelis et al., 2005. |
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.