Last data update: 24 January 2024 16:39 CET
Plasmid name: pGALMFHIFNB2 (LMBP 2473)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p2473.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Saccharomyces cerevisiae α-mating factor 1 gene (MFα1); prepro secretion signal sequence (ppMF) Human interferon β cDNA (IFNB); mature sequence |
| Promoter: | Saccharomyces cerevisiae galactokinase promoter (GAL1) 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: | pGALMFHIFNB1 |
| Further information: | This plasmid was derived from pGALMFHIFNB1 by introducing 4 extra codons encoding Glu-Ala-Glu-Ala (EAEA) between the prepro secretion signal sequence (ppMF) of the Saccharomyces cerevisiae α-mating factor 1 gene (MFα1) and the mature human interferon β gene (hIFNB) via insertion mutagenesis (Morinaga et al. (1984)) using the synthetic oligonucleotide 5'-GGATAAAAGGGAAGCTGAAGCTATGAGCTAC-3'. The hIFNB gene is placed under control of the inducible S. cerevisiae galactokinase promoter (GAL1). Attention: the vector part of the plasmid is probably not entirely correct: the restriction sites were not all experimentally verified. 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 at the 3' end of the GAL1 promoter is not exactly known. Other name of the plasmid is pSCGMFISHIFNB. |
| EMBL Accession number: | - |
| Latest sequence update: | 09/06/1994 |
| Sequence detail: | Nucleotide sequence at the fusion of the prepro secretion signal sequence (ppMF) of the Saccharomyces cerevisiae alpha-mating factor 1 gene (MFα1) to the mature human interferon ß sequence (hIFNB).
ppMF -----> -- mature hIFNB ->
5' ... GGG.GTA.TCT.TTG.GAT.AAA.AGG.GAA.GCT.GAA.GCT.ATG.AGC.TAC ... 3'
Gly Val Ser Leu Asp Lys Arg Glu Ala Glu Ala Met Ser Tyr
---------------
EAEA insert
Punctuation indicates reading frame. |
| 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. Contreras(1) (2). (1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium (2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | - |
| Related plasmid reference: | Morinaga et al., Biotechnology 2 (1984), 636-639 |
| Restricted distribution: | - BCCM MTA |
| Distributed as: | H/P active culture or plasmid DNA |
| Host for distribution: | Escherichia coli K12 |
| Host reference: | - |
| 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: |
pGALMFHIFNB2 (LMBP 2473) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Contreras . |
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.