Last data update: 24 January 2024 16:39 CET
Plasmid name: pSV23SmTNFd1-9K11E (LMBP 4638)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p4638.gb
(View with Genome Compiler) p4638.txt p4638.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Mouse tumor necrosis factor cDNA (Tnf, TNF); with deletion and mutation |
| Promoter: | 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: | pSV23SMTNF; pMcmTNFd1-9K11E |
| Further information: | This plasmid was constructed by inserting the SmaI-SstI (SacI) fragment of pMcmTNFd1-9K11E, containing mouse tumor necrosis factor cDNA with deletion and mutation (mTNFΔ1-9K11E), into the StuI-SstI (SacI) sites of pSV23SMTNF. The mTNFΔ1-9K11E cDNA lacks the first nine codons of the mature sequence, and has codon 11, coding for Lys, mutated to Glu. The mTNF mutant also contains a FspI (MstI) site due to silent mutations of codons 13 and 14. pSV23SmTNFd1-9K11E is an expression plasmid for uncleavable, membrane-bound TNF production. The biological activity of mTNFΔ1-9K11E is similar to that of wild type mTNF. The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727136.1. The nucleotide sequence of the wild type mTNF cDNA corresponds with the EMBL Nucleotide Sequence Database accession number M11731.1. The 5' and the 3' borders of the insert have been sequenced by LMBP. Name mentioned in Decoster et al. (1995) is pSV23SmTNFΔ1-9K11E. |
| EMBL Accession number: | M11731.1, view at EMBL, GenBank, DDBJ LT727136.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 27/11/2007 |
| Sequence detail: | Nucleotide sequence of mTNF cDNA at the start of the mature sequence:
mTNF wild type:
preseq.-->|--> mature
-1 | 1 2 3 4 5 6 7 8 9 10 11 12 13 14
5' ACC.CTC.ACA.CTC.AGA.TCA.TCT.TCT.CAA.AAT.TCG.AGT.GAC.AAG.CCT.GTA.GCC.CA 3'
----------------------------------- Lys
Mutated mTNF with 1-9 deletion:
preseq.-->|--> mature
-1| 10 11 12 13 14
5' ACC.CTC.ACA.GAC.GAG CCT.GTT.GCG.CA 3'
* * *
Glu FspI
*: mutated nucleotide.
---: deleted fragment. |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BamHI, BglI, EcoRI, FspI, SalI and XcmI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Dr E. Decoster(1) (2) and Prof. Dr W. Fiers(1) (2). (1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium (2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | Decoster et al., J. Biol. Chem. 270 (1995), 18473-18478 [PMID: 7629174] |
| Related plasmid reference: | Meszaros et al., J. Immunol. 165 (2000), 435-441 [PMID: 10861082] |
| 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: | LMBP 03404 |
Refer in your Materials and Methods: |
pSV23SmTNFd1-9K11E (LMBP 4638) is available at BCCM/GeneCorner. This plasmid was deposited by Dr E. Decoster and Prof. Dr W. Fiers and was published in Decoster et al., 1995. |
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.