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GeneCorner plasmid details

Last data update: 24 January 2024 16:39 CET

Plasmid name: pGAPNorMNN4B (LMBP 11112)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p11112.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Pichia pastoris Mnn4Bp cDNA (MNN4B)
Promoter: Pichia pastoris glyceraldehyde-3-phosphate dehydrogenase promoter (GAP)
Saccharomyces cerevisiae translation elongation factor 1α promoter (TEF1α)
Synthetic prokaryotic EM7 promoter
Ribosome
binding site:
-
Terminator: Saccharomyces cerevisiae iso-1-cytochrome C terminator (CYC1)
Pichia pastoris alcohol oxidase 1 terminator (AOX1)
Selection marker: Nourseothricin (ntc)
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli
Pichia pastoris; integrative
Parental clone: pGAPNor
Further information: The plasmid was constructed by PCR amplifying the P. pastoris MNN4B coding sequence from P. pastoris GS115deltaOCH1, and cloning it into the pGAPNor vector.
The plasmid is intended for overexpression of P. pastoris MNN4B after chromosomal integration of the linearised plasmid.
The nucleotide sequence of the P. pastoris MNN4B coding sequence corresponds with the EMBL Nucleotide Sequence Database accession number FN392320.1.
Other name of the plasmid is pGAPNOR-MNN4B-cloneEZ.
EMBL Accession number: FN392320.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 11/09/2018
Sequence detail:
Primers used to amplify the P. pastoris MNN4B coding sequence: 

Forward: 5' TGTCCCTATTTCAATCAATTGAACAACTAT.ATG.TTC.AAA.GAA.ACG.TCA.AAG.AAC.TTG
                                           ***

Reverse: 5' ATGATGATGATGATGATGGTCGAC.GTC.GTT.GTG.TTC.AAG.TTT.GAC

***: Starting codon
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 N. Callewaert(1) (2).
(1) VIB-UGent Center for Medical Biotechnology, VIB, Ghent, Belgium
(2) Department of Biochemistry and Microbiology, Ghent University, Ghent, Belgium
Plasmid reference: PhD thesis Charlotte De Visscher (2015)
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 DH5α
Host reference: Focus 8 (1986), 9
Related host reference: Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660]
Rodriguez-Quinones et al., Focus 15 (1993), 110-112
Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791] [DOI: 10.1016/s0022-2836(83)80284-8]
Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187]
Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051]
Cultivation medium: LB-Lennox + nourseothricin (50 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
Other culture collection numbers: -

Refer in your Materials and Methods:

pGAPNorMNN4B (LMBP 11112) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr N. Callewaert and was published in Charlotte De Visscher, 2015.

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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