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LMBP plasmid details

Last data update: 24 November 2020 04:18 CET

Plasmid name: pPLcMIFNG (LMBP 1598)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p1598.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Mouse interferon γ cDNA (IFNG); (slightly modified) 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: -
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: pPLc321; pATMIFNG
Further information: The plasmid was constructed as follows: (i) the MaeI-Sau3AI fragment (mouse mature IFNG fragment + 3UTR) from pATMIFNG was isolated, and ligated with a synthetic sequence (26 bp) at the MaeI site (restoring 7 of the first 10 amino acids of the mature mouse IFNG CDS and adding a NcoI compatible sequence);(ii) this fragment was inserted between the NcoI and BamHI sites of pPLc321.
pPLcMFNG is an expression plasmid, designed for PL-controlled expression of mature mouse IFNG.
The length of the 3' untranslated region of the mouse IFNγ insert is not completely known.
The nucleotide sequence of the mouse IFNG cDNA corresponds with the EMBL Nucleotide Sequence Database accession number K00083.1, except for the slightly modified 5' side.
Other name of the plasmid is pPLcMIFNG15.
EMBL Accession number: K00083.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 26/05/1989
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 

                                    Met His Gly Thr Val Ile Glu Ser Leu Glu Ser Leu
pPLcMIFNG: 5'...TAAAAAGGGTATCGATTCC.ATG.CAC.GGC.ACA.GTC.ATT.GAA.AGC.CTA.GAA.AGT.CTG...3'
                          ClaI    -----------------------------------
                                           linker                   MaeI
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: -
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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