Last data update: 24 January 2024 16:39 CET
Plasmid name: pPpT4-NSS-TrCel61A-Y24A-Y211A (LMBP 12111)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | not available |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Trichoderma reesei cellulase 61A cDNA (Cel61A, EGIV, egl4, endoglucanase-4); mutated sequence 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-NSS-TrCel61A-CO |
| Further information: | The plasmid was constructed by introducing Y24A and Y211A amino acid mutations in the T. reesei Cel61A coding sequence of pPpT4-NSS-TrCel61A-CO. Subsequently, the plasmid was linearised and integrated in the P. pastoris genomic DNA. The T. reesei Cel61A coding sequence is preceded by its native secretion signal and was codon-optimised for P. pastoris. The Y24A and Y211A mutations were introduced to study regioselectivity. The nucleotide sequence of the wild type 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 Pichia-NSS-TrCel61A-Y24A-Y211A (sPichExtra108). |
| EMBL Accession number: | Y11113.1, view at EMBL, GenBank, DDBJ JQ519689.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 05/06/2020 |
| 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 Z. Ubiparip(1) and Prof. Dr T. Desmet(1). It was constructed by Dr B. Danneels(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: | active culture |
| Host for distribution: | Pichia pastoris |
| Host reference: | - |
| Cultivation medium: | YPD + zeocin (s: 100 μg/ml; l: 10 μg/ml) |
| Cultivation temperature: | 28°C |
| Biosafety level: | L1 |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pPpT4-NSS-TrCel61A-Y24A-Y211A (LMBP 12111) is available at BCCM/GeneCorner. The plasmid was deposited by Z. Ubiparip and Prof. Dr T. Desmet and was published in Tanghe et al., 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.