Last data update: 24 January 2024 16:39 CET
Plasmid name: pGAPNorPNO1 (LMBP 11106)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p11106.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Pichia pastoris RNA-binding protein Pno1p cDNA (PNO1) |
| 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: | Pichia pastoris alcohol oxidase 1 terminator (AOX1) Saccharomyces cerevisiae iso-1-cytochrome C terminator (CYC1) |
| 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 PNO1 coding sequence from P. pastoris GS115deltaOCH1, and cloning it into the pGAPNor vector. The plasmid is intended for overexpression of P. pastoris PNO1 after chromosomal integration of the linearised plasmid, providing extra mannophosphorylation on glycans. The nucleotide sequence of the P. pastoris PNO1 coding sequence corresponds with the EMBL Nucleotide Sequence Database accession number FN392320.1. Other name of the plasmid is pGAPNOR-PNO1-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 PNO1 coding sequence:
Forward: 5' TGTCCCTATTTCAATCAATTGAACAACTAT.ATG.ACA.CTC.AGG.AGT.GCC.ATA.AAA.G
***
Reverse: 5' ATGATGATGATGATGATGGTCGAC.CTG.TTT.TCT.ATG.CTT.AGA.AAG.GTC
***: 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: | Tiels et al., Nat. Biotechnol. 30 (2012), 1225-1231 [PMID: 23159880] [DOI: 10.1038/nbt.2427] |
| Related 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: |
pGAPNorPNO1 (LMBP 11106) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr N. Callewaert and was published in Tiels et al., 2012. |
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.