Last data update: 24 January 2024 16:39 CET
Plasmid name: pPIC9-hLAL (LMBP 11124)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p11124.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human lipase A, lysosomal acid type cDNA (LIPA, CESD, LAL, GeneID 3988); mature 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; his4(-), integrative |
| Parental clone: | pPIC9 |
| Further information: | The plasmid was constructed by cloning the human LIPA mature sequence as an SpeI/NotI fragment into the AvrII/NotI opened pPIC9 vector. The plasmid is meant to overexpress human LIPA in Pichia pastoris. The human LIPA coding sequence does not contain a signal sequence. 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 human LIPA coding sequence corresponds with the EMBL Nucleotide Sequence Database accession number M74775.1. Other name of the plasmid is pPic9_hLAL. |
| 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: | 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 + ampicillin (100 μg/ml) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pPIC9-hLAL (LMBP 11124) 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.