Last data update: 24 January 2024 16:39 CET
Plasmid name: pSP65HTNF (LMBP 2922)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p2922.gb
(View with Genome Compiler) p2922.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human tumor necrosis factor cDNA (TNF) |
| Promoter: | Phage SP6 promoter Escherichia coli lac operon promoter |
| Ribosome binding site: |
- |
| Terminator: | - |
| Selection marker: | Ampicillin (amp) |
| Replicon: | Escherichia coli plasmid pMB1 origin |
| Host range: | Escherichia coli |
| Parental clone: | pSP65; pATHTNF |
| Further information: | This plasmid was constructed as follows: 1) the FokI fragment of pATHTNF containing the human tumor necrosis factor (hTNF) coding sequence (except the termination codon) was blunted (Klenow exonuclease) and inserted in the unique SmaI site of the polylinker of pSP65; 2) To complete the coding sequence, the ApaI-DraI (AhaIII) fragment of pATHTNF (position 228-1511 of the hTNF cDNA) was used to replace the ApaI-HindII fragment of the intermediary construct. 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 LT727558.1. |
| EMBL Accession number: | LT727558.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 08/07/1993 |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: EcoRI, HindII, HindIII and RsaI. Referring to the expected fragment sizes deduced from the compiled nucleotide sequence, the presence, the length and the orientation of the hTNF insert could be confirmed, but the vector part of the plasmid seems to be larger. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by 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: | Muller et al., FEBS Lett. 197 (1986), 99-104 [PMID: 3512310] |
| Restricted distribution: | - BCCM MTA |
| Distributed as: | H/P active culture or plasmid DNA |
| Host for distribution: | Escherichia coli K12 RR1ZΔM15 |
| Host reference: | Altieri et al., J. Bacteriol. 168 (1986), 648-654 [PMID: 2946661] |
| Cultivation medium: | LB-Lennox + ampicillin (100 μg/ml) |
| Cultivation temperature: | 28°C |
| Biosafety level: | L1 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pSP65HTNF (LMBP 2922) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr W. Fiers and was published in Muller et al., 1986. |
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.