Last data update: 24 January 2024 16:39 CET
Plasmid name: pPLcMIFNG3 (LMBP 1939)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p1939.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Mouse interferon γ cDNA (IFNG); secretion variant of the mature sequence |
| Promoter: | Phage λ major leftward promoter (λ PL) |
| Ribosome binding site: |
Ribosome binding site (RBS) of the Escherichia coli tryptophan operon attenuator peptide (trp) |
| Terminator: | Phage T4 gene 32 terminator (T4g32) |
| Selection marker: | Ampicillin (amp) |
| Replicon: | Escherichia coli plasmid pMB1 origin |
| Host range: | Escherichia coli; use strains with a cI function, cIts for PL controlled expression |
| Parental clone: | pPLcMIFNG; pPLc28 |
| Further information: | The plasmid was constructed as follows: (i) the ClaI-OxaNI fragment (mouse IFNG) was isolated from pPLcMIFNG and ligated with a synthetic sequence (24 bp) at the OxaNI (=MstII) site (replacing 5 of the 7 terminal codons of the mouse IFNG gene and adding a BamHI site);(ii) this fragment was inserted in the unique ClaI and BamHI sites of pPLc28HIIF-52T4. pPLcMFNG3 is an expression plasmid, designed for PL-controlled expression of mature mouse IFNG. It was intended for periplasmic secretion. This linker results in the deletion of the Arg-Cys residues at the carboxyl end of mouse IFNG. The nucleotide sequence of the mouse IFNG cDNA corresponds with the EMBL Nucleotide Sequence Database accession number K00083.1, except for the modified 3' and 5' side. Other name of the plasmid is pPLcMIFNG24-T4. |
| EMBL Accession number: | K00083.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 19/10/1989 |
| Sequence detail: | signal sequence ->|-> mature mouse IFNG
Val Ser Gly Cys Tyr Cys His Gly Thr Val Ile Glu Ser Leu Glu Ser Leu
wild type: 5'...GTT.TCT.GGC|TGT.TAC.TGC.CAC.GGC.ACA.GTC.ATT.GAA.AGC.CTA.GAA.AGT.CTG...3'
MaeI
Glu Ser Ser Leu Arg Lys Arg Lys Arg Ser Arg Cys |-> 3'UTR of mouse IGNG
...GAA.TCC.AGC.CTC.AGG.AAG.CGG.AAA.AGG.AGT.CGC.TGC.TGA.TTCGGGGTGGGGA...
OxaNI
Met His Gly Thr Val Ile Glu Ser Leu Glu Ser Leu
pPLcMIFNG3: 5'...TAAAAAGGGTATCGATTCC.ATG.CAC.GGC.ACA.GTC.ATT.GAA.AGC.CTA.GAA.AGT.CTG...3'
-----------------------------------
linker MaeI
Glu Ser Ser Leu Arg Lys Arg Lys Arg Ser |-> T4 terminator
...GAA.TCC.AGC.CTC.AGG.AAG.CGT.AAA.CGT.TCA.TGA.GGATCCGGTGGATG...
OxaNI BamHI |
| 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 E. Remaut(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: | Simons et al., Gene 28 (1984), 55-64 [PMID: 6234201] |
| Restricted distribution: | - BCCM MTA |
| Distributed as: | H/P active culture or plasmid DNA |
| Host for distribution: | Escherichia coli K12 MC1061[pcI857] |
| Host reference: | Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493] |
| Helper plasmid: | pcI857 |
| Cultivation medium: | LB-Lennox + ampicillin (100 μg/ml) + kanamycin (50 μg/ml) |
| Cultivation temperature: | 28°C |
| Biosafety level: | L1 |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pPLcMIFNG3 (LMBP 1939) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr E. Remaut . |
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.