Last data update: 24 January 2024 16:39 CET
Plasmid name: pSVSPORTcHLhTNFE (LMBP 3796)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p3796.gb
(View with Genome Compiler) p3796.txt p3796.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Gallus gallus hepatic lectin cDNA (HL, GeneID 396477); presequence Human tumor necrosis factor cDNA (TNF); mature sequence E-tag; C-terminal |
| Promoter: | Phage SP6 promoter Phage T7 gene 10 promoter (T7g10) Escherichia coli lac operon promoter; mutant (lacUV5) 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: | pSV-Sport-Etag |
| Further information: | The HL-TNF fusion gene was constructed by fusion PCR and inserted as an EcoRI-NotI fragment into the EcoRI-NotI opened pSV-Sport-Etag vector. As a result, the E-tag is C-terminally fused to the fusion gene. This plasmid contains the chicken hepatic lectin membrane anchor sequence fused to the E-tagged, human TNF mature cDNA. When cloning a fragment downstream from the lac promoter it may be advisable to use lacI(q) strains in order to prevent fortuitous expression of a possibly noxious polypeptide. The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727394.1. The nucleotide sequence of the human TNF mature sequence corresponds with the EMBL Nucleotide Sequence Database accession number X01394.1. Other names of the plasmid are pSVSPORT1.CHL.HTNF-Etag and pSV-SPORT-1.CHL.HTNF E-tag. |
| EMBL Accession number: | X01394.1, view at EMBL, GenBank, DDBJ LT727394.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 31/01/2000 |
| Sequence detail: | Nucleotide sequence at the end of the fusion gene:
hTNF --> --> E-tag -->
5' GCC.CTG.GCG.GCC.GCA.GGT.GCG.CCG.GTG.CCG.TAT.CCG.GAT.CCG.CTG.GAA.CCG.CGT.GCC.GCA.TAG 3'
NotI +++
+++: termination codon.
Punctuation indicates reading frame. |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BanI, EcoRI, HindII, KpnI/XbaI, NotI/SspI, PstI and XmnI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Dr E. Decoster(1) (2) and 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: | Decoster et al., J. Cell Biol. 143 (1998), 2057-2065 [PMID: 9864375] |
| 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: |
pSVSPORTcHLhTNFE (LMBP 3796) is available at BCCM/GeneCorner. This plasmid was deposited by Dr E. Decoster and Prof. Dr R. Beyaert 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.