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LMBP plasmid details

Last data update: 24 November 2020 04:18 CET

Plasmid name: pPIC9-sspLAL (LMBP 11122)

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

-

Cloned DNA: Human lipase A, lysosomal acid type cDNA (LIPA, CESD, LAL, GeneID 3988); codon optimised sequence
Saccharomyces cerevisiae α-mating factor 1 gene (MFα1); prepro secretion signal sequence (ppMF)
Histidine tag (His-tag); C-terminal
Promoter: Pichia pastoris alcohol oxidase 1 promoter (AOX1)
Ribosome
binding site:
-
Terminator: Pichia pastoris alcohol oxidase 1 terminator (AOX1)
Selection marker: Ampicillin (amp)
Pichia pastoris HIS4; auxotrophic
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli
Pichia pastoris
Parental clone: pPIC9
Further information: The plasmid was constructed by cloning the LIPA coding sequence, codon optimised for P. pastoris, as an AvrII/NotI fragment into the AvrII/NotI opened pPIC9 vector.
The plasmid is meant to overexpress codon optimised LIPA in Pichia pastoris.
The presence of the P. pastoris HIS4 gene and two P. pastoris AOX1 regions (the AOX1 promoter and the AOX1 3' flanking region) allows integration of the linearized vector into the Pichia genome via homologous recombination at the his4 locus or AOX1 locus respectively.
The nucleotide sequence of the wild type human LIPA coding sequence corresponds with the EMBL Nucleotide Sequence Database accession number M74775.1.
Other name of the plasmid is pPic9_sspLAL.
EMBL Accession number: M74775.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 06/09/2018
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)
Related plasmid reference: Cregg et al., Mol. Cell. Biol. 9 (1989), 1316-1323 [PMID: 2657390]
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: Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051]
Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187]
Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791]
Rodriguez-Quinones et al., Focus 15 (1993), 110-112
Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660]
Cultivation medium: LB-Lennox + ampicillin (100 μg/ml)
Cultivation temperature: 37°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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