Last data update: 24 January 2024 16:39 CET
Plasmid name: pPTK079_P8_Lox71-ZeoR-Lox66 (LMBP 12793)
New search | Print data sheet |
Price category: | Cat. 3 (cf. price list) |
Status: | GeneCorner core plasmid |
GeneCorner sequence: |
p12793.gb
(View with Genome Compiler) p12793.txt p12793.pdf |
Sequence analysis results Genecorner: |
Sanger: ef32243454.fasta Sanger: ef32708293.fasta |
Cloned DNA: |
Discosoma sp. red fluorescent protein DNA (DsRed1); mutated monomeric variant (mRFP1) |
Promoter: | Escherichia coli tetracycline inducible promoter (PLtetO-1) Pichia ketol-acid reductoisomerase promoter (ILV5) Synthetic prokaryotic EM72 promoter |
Ribosome binding site: |
Ribosome binding site (RBS); synthetic sequence |
Terminator: | Escherichia coli rrnB operon T1 terminator Phage T7 early transcription terminator (T7Te) Eremothecium gossypii translation elongation factor 1α terminator (TEF1α) |
Selection marker: | Bleomycin (bleo; zeomycin (zeo; Zeocin); phleomycin (phleo)) |
Replicon: | Escherichia coli plasmid pMB1 origin |
Host range: | Escherichia coli Pichia pastoris; as part of the OPENPichia plasmid kit |
Parental clone: | pPTK_8 |
Further information: | The plasmid was constructed by cloning the zeocin resistance cassette, flanked by Lox71 and Lox66 sites, in the synthetic pPTK_8 vector. This plasmid is a level 1 destination vector (OPENPichia part 8) for the OPENPichia modular cloning technology. OPENPichia parts 1 to 8 can be combined via BsaI restriction and ligation of the parts into a Pichia expression plasmid. Alternatively, a transcriptional unit, consisting of OPENPichia parts 1 to 5 can be assembled via BsaI restriction and ligation. Several transcriptional units can be combined in a multigene destination vector via BsmBI restriction and ligation. The PLtetO-1 promoter has 2 tetO operators and can be repressed by the Tet repressor (TetR) or activated by anhydrotetracycline (aTc). The zeocin resistance cassette is flanked by two single-mutant LoxP sites (Lox71 and Lox66), and can be removed by Cre recombinase, leaving a double mutant LoxP site (Lox72). Other name of the plasmid is pPTK079. |
EMBL Accession number: | - |
Latest sequence update: | 07/05/2021 |
Authenticity test: | Restriction enzyme pattern analysed at BCCM/GeneCorner: HindIII, NcoI and PvuII. The mRFP1 gene was sequenced at BCCM/GeneCorner. The plasmid has also been fully sequenced. |
Class: | Recombinant plasmid |
Type: | Plasmid |
History of deposit: | This plasmid was deposited by S. Vanmarcke(1)(2) and Prof. Dr N. Callewaert(1)(2). It was constructed by Dr K. Vandewalle(1)(2) and Dr R. Vanluchene(1)(2). (1) VIB-UGent Center for Medical Biotechnology, VIB, Ghent, Belgium (2) Department of Biochemistry and Microbiology, Ghent University, Ghent, Belgium |
Plasmid reference: | Claes et al., Nature (2024) [PMID: 38443579] [DOI: 10.1038/s41564-023-01574-w] |
Restricted distribution: | - BCCM MTA adapted for OPENPichia - RECIPIENT agrees to refer to the 'Plasmid reference' in the first publication making use of the MATERIAL. - When appropriate in accordance with academic customs, RECIPIENT agrees to include the depositor(s) as co-author(s) in the first publication making use of the MATERIAL. - When making use of the MATERIAL, RECIPIENT agrees to refer to ‘OPENPichia’ in the Materials and Methods section and in the acknowledgments of all publications, as well as when launching commercial products. - The depositor will be informed of the customer's identity upon release of a sample outside the depositor's department. |
Distributed as: | H/P active culture or plasmid DNA |
Host for distribution: | Escherichia coli K12 DH5αT1R |
Host reference: | - |
Related host reference: | Killmann et al., J. Bacteriol. 178 (1996), 6913-6920 [PMID: 8955314] [DOI: 10.1128/jb.178.23.6913-6920.1996] |
Cultivation medium: | LB-Lennox + zeocin (25 μg/ml) |
Cultivation temperature: | 28°C |
Biosafety level: | L1 |
Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pPTK079_P8_Lox71-ZeoR-Lox66 (LMBP 12793) is available at BCCM/GeneCorner. This plasmid was deposited by S. Vanmarcke and Prof. Dr N. Callewaert and was published in Claes et al., 2024. |
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.