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LMBP plasmid details

Last data update: 24 November 2020 04:18 CET

Plasmid name: pPLcMIFNG3 (LMBP 1939)

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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: -

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.

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