Last data update: 24 January 2024 16:39 CET
Plasmid name: pSV23SmIL6mTNF (LMBP 3409)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p3409.gb
(View with Genome Compiler) p3409.txt p3409.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Mouse interleukin 6 cDNA (Il6, Il-6, GeneID 16193); signal sequence with silent mutation Mouse tumor necrosis factor cDNA (Tnf, TNF); mature sequence |
| 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: | pSV23SmIL6m; pSV23SmTNFm3 |
| Further information: | This plasmid was constructed as follows: the BglII-SalI fragment of pSV23SmTNFm3, containing the mature sequence of the mTNF cDNA, was ligated to the large StuI/SalI fragment of pSV23SmIL6m by insertion of a synthetic linker. pSV23SmIL6mTNF contains the mutated signal sequence of the mIL6 cDNA fused in frame to the mature mTNF sequence. The mutation of the mIL6 signal sequence exists of a silent mutation of codon -10. Expression of the mIL6-mTNF chimeric gene gives rise to secreted mTNF. The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727482.1. Other name of the plasmid is pSV23SmIL6.TNF. |
| EMBL Accession number: | M11731, view at EMBL, GenBank, DDBJ LT727482.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 16/04/1996 |
| Sequence detail: | Fusion region between the mutated mIL6 signal sequence and the mTNF mature sequence:
mIL6 signal sequence ->|-> mTNF mature sequence
-10 -1| 1 2 3 4
5' ... TTC.TTA.GGA.CTG.ATG.CTG.GTG.ACA.ACC.ACG.GCC.CTC.AGA.TCT.TCT ... 3'
* --------------------------------------- *
^ ^^ BglII
---: Linker.
*: Mutated nucleotide.
^: Rest of StuI from pSV23smIL6m.
Nucleotide sequence of the synthetic linker:
5' A.CTG.ATG.CTG.GTG.ACA.ACC.ACG.GCC.CTC.A 3'
3' T.GAC.TAC.GAC.CAC.TGT.TGG.TGC.CGG.GAG.TCT.AG 5'
BglII |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BamHI, BglI/XmnI, BglII/SalI, EcoRI/PstI, PvuII/StuI and TaqI. The pattern revealed that approx. 60 nucleotides are missing from the 3' untranslated region of the mTNF 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) and Prof. Dr W. Fiers(1). (1) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | Decoster et al., J. Biol. Chem. 273 (1998), 3271-3277 [PMID: 9452442] |
| 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: |
pSV23SmIL6mTNF (LMBP 3409) 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., 1998. |
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.