Last data update: 24 January 2024 16:39 CET
Plasmid name: pSV23SmTNFd1-12 (LMBP 3406)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p3406.gb
(View with Genome Compiler) p3406.txt p3406.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Mouse tumor necrosis factor cDNA (Tnf, TNF); with deletion |
| 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; pMcmTNFd1-12 |
| Further information: | This plasmid was constructed as follows: 1) deletion of the first twelve codons of the mature mouse TNF sequence was performed via site-directed mutagenesis in the pMaMTNF vector, resulting in pMcmTNFd1-12; 2) the mutated mouse TNF gene was then inserted as a SmaI-SstI(SacI) fragment into the StuI-SstI(SacI) sites of pSV23SMTNF, resulting in the loss of the StuI site. pSV23SmTNFd1-12 is an expression plasmid for an uncleavable, membrane bound form of mouse TNF. This mouse TNF variant is less potent than mTNFΔ1-9K11E. The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727480.1. The nucleotide sequence of the wild type mouse TNF corresponds with the EMBL Nucleotide Sequence Database accession number M11731.1. Name mentioned in Decoster et al. (1995) is pSV23SmTNFΔ1-12. |
| EMBL Accession number: | M11731.1, view at EMBL, GenBank, DDBJ LT727480.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 31/01/1996 |
| Sequence detail: | Mouse TNF at the start of the mature sequence:
mouse TNF wild type:
preseq.->|-> mature
-1 | 1 2 3 4 5 6 7 8 9 10 11 12 13
5' ACC.CTC.ACA.CTC.AGA.TCA.TCT.TCT.CAA.AAT.TCG.AGT.GAC.AAG.CCT.GTA.G 3'
-----------------------------------------------
mouse TNF with 1-12 deletion:
preseq.->|-> mature
-1| 13 14
5' ACC.CTC.ACA.GTA.GCC. 3'
---: deleted fragment.
Punctuation indicates reading frame. |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: EcoRI, HaeII/StuI, RsaI/XhoI, SacI/SmaI, SalI and TaqI. The pattern revealed that approx. 60 nucleotides are missing from the 3' untranslated region of the mouse TNF gene, including the 3'-terminal (supposedly reconstructed) EcoRI site. |
| 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] |
| 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: |
pSV23SmTNFd1-12 (LMBP 3406) 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.