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LMBP plasmid details

Last data update: 24 November 2020 04:18 CET

Plasmid name: pPpT4-NSS-TrCel61A-CO (LMBP 10347)

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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: Trichoderma reesei cellulase 61A cDNA (Cel61A, EGIV, egl4, endoglucanase-4)
Histidine tag (His-tag); C-terminal
Promoter: Pichia pastoris alcohol oxidase 1 promoter (AOX1)
Synthetic prokaryotic EM72 promoter
Pichia ketol-acid reductoisomerase promoter (ILV5)
Ribosome
binding site:
-
Terminator: Pichia pastoris alcohol oxidase 1 terminator (AOX1)
Pichia pastoris alcohol oxidase terminator (AOD)
Selection marker: Bleomycin (bleo; zeomycin (zeo; Zeocin); phleomycin (phleo))
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli
Pichia pastoris; integrative
Parental clone: pPpT4
Further information: The plasmid was constructed by PCR amplifying the T. reesei Cel61A coding sequence and cloning it into the E. coli - P. pastoris shuttle vector pPpT4.
The T. reesei Cel61A coding sequence is preceded by its native secretion signal and was codon-optimised for P. pastoris.
The plasmid is also available for expression in P. pastoris, integrated in the genomic DNA, under accession number LMBP 12112.
The nucleotide sequence of the T. reesei Cel61A coding sequence corresponds with the EMBL Nucleotide Sequence Database accession number Y11113.1.
The nucleotide sequence of the pPpT4 parental vector corresponds with the EMBL Nucleotide Sequence Database accession number JQ519689.1 except for some nucleotides upstream of the AOX1 promoter.
Other name of the plasmid is TrCel61A-CO-NSS_CO -- COLI.
EMBL Accession number: Y11113.1, view at EMBL, GenBank, DDBJ
JQ519689.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 24/02/2017
Authenticity test: The plasmid still needs to be subjected to the authenticity test.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: The plasmid was deposited by Prof. Dr T. Desmet(1). It was constructed by M. Tanghe(1).
(1) Center for Synthetic Biology, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium
Plasmid reference: Tanghe et al., Mol Biotechnol. 57 (2015), 1010-1017 [PMID: 26285758] [DOI: 10.1007/s12033-015-9887-9]
Related plasmid reference: Danneels et al., Biotechnol J 14 (2019), e1800211 [PMID: 30238672] [DOI: 10.1002/biot.201800211]
Restricted distribution: - BCCM MTA
- The depositor will be informed of the customer's identity upon release of a sample outside the depositor's department or outside the departments in which BCCM/GeneCorner is embedded, namely UGent-DBMB and VIB-IRC.
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 + zeocin (25 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
Other culture collection numbers: LMBP 12112

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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