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GeneCorner plasmid details

Last data update: 24 January 2024 16:39 CET

Plasmid name: pMC_Cre_NourR (LMBP 12799)

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Price category: Cat. 3 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p12799.gb (View with Genome Compiler)
p12799.txt
p12799.pdf
Sequence
analysis results
Genecorner:

Sanger: ef32243536.fasta

Sanger: ef32243541.fasta

NGS: 12799d-ps-gc-jul2023.fasta

NGS: 12799k-ps-gc-jul2023.fasta

Cloned DNA: Phage P1 cre gene encoding recombinase
Modular cloning left assembly connector S (ConLS)
Modular cloning right assembly connector 1 (ConR1)
Promoter: Pichia ketol-acid reductoisomerase promoter (ILV5)
Synthetic prokaryotic EM72 promoter
Saccharomyces phosphopyruvate hydratase ENO1 promoter (ENO1)
Ribosome
binding site:
-
Terminator: Eremothecium gossypii translation elongation factor 1α terminator (TEF1α)
Pichia pastoris alcohol oxidase 1 terminator (AOX1)
Selection marker: Ampicillin (amp)
Nourseothricin (ntc)
Replicon: Escherichia coli plasmid pMB1 origin
Pichia pastoris autonomously replicating sequence (ARS1)
Host range: Escherichia coli
Pichia pastoris
Parental clone: pPTK070_P1_ConLS; pPTK015_P2_pENO1; pPTK_P3_Cre; pPTK047_P4_AOX1tt; pPTK073_P5_ConR1; pPTK004_P7_Stuffer; pPTK080_P8_AmpR; pPTK201_P6_NourR
Further information: The plasmid was constructed by BsaI restriction on pPTK070_P1_ConLS, pPTK015_P2_pENO1, pPTK_P3_Cre, pPTK047_P4_AOX1tt, pPTK073_P5_ConR1, pPTK201_P6_NourR, pPTK004_P7_Stuffer and pPTK080_P8_AmpR, and ligating the OPENPichia parts together.
This plasmid is a level 1 multigene destination vector for the OPENPichia modular cloning technology.
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.
EMBL Accession number: -
Latest sequence update: 17/10/2023
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: EcoRI/PstI and PvuII.

The Cre gene and the region with the nourseothricin resistance gene, TEF1 terminator and ARS1 were 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 + ampicillin (100 µg/ml) + nourseothricin (50 µg/ml)
Cultivation temperature: 28°C
Biosafety level: L1
Other culture collection numbers: -

Refer in your Materials and Methods:

pMC_Cre_NourR (LMBP 12799) 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.

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