Last data update: 24 January 2024 16:39 CET
Plasmid name: pSV23SgHLmTNF (LMBP 3415)
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Price category: | Cat. 1 (cf. price list) |
Status: | GeneCorner non-core plasmid |
Depositor's sequence: | p3415.gb |
Sequence analysis results Genecorner: |
- |
Cloned DNA: |
Gallus gallus hepatic lectin cDNA (HL, GeneID 396477) 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: | pSV23SmTNF; pMcmTNFmBglII; pCHL213 |
Further information: | This plasmid was constructed as follows: 1/ a BclI site was created by site-specific mutagenesis of codons 71 (silent) and 73 of the chicken hepatic lectin (gHL) gene in pMaCHL. 2/ The gHL-mTnf fusion gene was created by inserting the small EcoRI-BclI fragment of the pMaCHL mutant (containing the gHL anchor) and a synthetic linker (containing the 4 last codons of the presequence of mTnf) between the EcoRI and BglII sites of the pMcmTNFmBglII. 3/ The fusion gene was then inserted as a SacI-EcoRI fragment (filled-in with Klenow DNA polymerase) between the SacI and StuI sites of pSV23SMTNF. Restriction-enzyme digestions revealed that approx. 60 nucleotides are missing from the 3' UTR region of the mTnf gene, including the 3'-terminal (supposedly restored) EcoRI site. The plasmid contains the SV40 small t-antigen splicing signal. Other name of the plasmid is pSV23SCHL.TNF. |
EMBL Accession number: | M11731, view at EMBL, GenBank, DDBJ |
Latest sequence update: | 11/03/1996 |
Sequence detail: | Fusion region between the gallus gallus (chicken) hepatic lectin anchor and mature, murine Tnf: | mTnf gHL | presequence | mature mTnf -70 | -4 -3 -2 -1| 1 2 3 4 5 6 ...TTC.TCC.AGC.CGT.GAT.CAG.ACC.CTC.ACA.CTC.AGA.TCT.TCT.TCT.CAA... * * BclI BglII * : mutated nucleotide linker sequence : 5' GAT.CAG.ACC.CTC.ACA.CTC.A 3' 3' TC.TGG.GAG.TGT.GAG.TCT.AG 5' BclI BglII |
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 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: | - |
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 |
Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pSV23SgHLmTNF (LMBP 3415) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Beyaert . |
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.