Last data update: 24 January 2024 16:39 CET
Plasmid name: pSV23SgHLmTNFd (LMBP 3416)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p3416.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Gallus gallus hepatic lectin cDNA (HL, GeneID 396477) Mouse tumor necrosis factor cDNA (Tnf, TNF); mature sequence 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; pMacmTNFΔ1-9K11E; 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 was created by inserting the small EcoRI-BclI fragment of the pMaCHL mutant (containing the gHL anchor) and a synthetic linker between the EcoRI and FspI sites of pMacmTNFΔ1-9K11E. 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. The mature mTnf sequence contains a deletion of amino acid codons 1 to 9 and a mutation of amino acid 11 from K to E. 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.TNFΔ1-9K11E. |
| EMBL Accession number: | M11731, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 12/03/1996 |
| Sequence detail: | Fusion region between the gallus gallus (chicken) hepatic lectin anchor
and mature, murine Tnf:
g HL | |mature mTnf with 1-9 deletion
-70 -69 -68 10 11 12 13 14
...TTC.TCC.AGC.CGT.GAT.CAG.GAC.GAG.CCT.GTT.GCG...
* * * *
Gln Glu
BclI FspI
* : mutated nucleotide
linker sequence : 5' GAT.CAG.GAC.GAG.CCT.GTT.GC 3'
3' TC.CTG.CTC.GGA.CAA.CG 5'
BclI FspI |
| 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: |
pSV23SgHLmTNFd (LMBP 3416) 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.