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GeneCorner plasmid details

Last data update: 24 January 2024 16:39 CET

Plasmid name: pET-DUET-hACLY-A/B (LMBP 11131)

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Price category: Cat. 3 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p11131.gb (View with Genome Compiler)
p11131.txt
p11131.pdf
Sequence
analysis results
Genecorner:

NGS: f11-gc-dec2018.fasta

Cloned DNA: Human ATP citrate lyase cDNA (ACLY, ATPCL, CLATP, ACL, GeneID 47); without linker
Escherichia coli lac repressor gene (lacI)
Histidine tag (His-tag); C-terminal
S-tag; C-terminal
Promoter: Phage T7 gene 10 promoter (T7g10) with lac operator (T7lac)
Escherichia coli lac repressor promoter (lacI)
Ribosome
binding site:
Ribosome binding site (RBS) of the phage T7 gene 10 (T7g10)
Terminator: Phage T7 gene 10 terminator (T7g10)
Selection marker: Ampicillin (amp)
Replicon: Escherichia coli plasmid pMB1 origin
Phage f1 origin
Host range: Escherichia coli
Parental clone: pETDuet-1
Further information: The plasmid was constructed by cloning the codon-optimized cDNA fragments encoding amino acids 1-425 (ACLY-A) and amino acids 487-1101 (ACLY-B) of human ACLY (uniprot ID P53396-1) into the pETDuet-1 vector.
This plasmid encodes for a heteromeric form of human ACLY without the long linker region (amino acids 426-486, following NM_001096.3).
The human ACLY-B fragment carries a C-terminal His-tag.
The nucleotide sequence of the wild type human ACLY coding sequence corresponds with the Genbank accession number NM_001096.3.
Other names of the plasmid are pETDuet1-hACLB-hACLA and 353_pETDuet-1_hACLB_hACLA.
EMBL Accession number: NM_001096.3, view at GenBank
Latest sequence update: 11/02/2019
Authenticity test: This plasmid has been fully sequenced.

The length of the plasmid has also been confirmed by PvuI restriction enzyme analysis.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: The plasmid was deposited by A. Dansercoer(1)(2), Dr K. Verstraete(1)(2) and Prof. Dr S. Savvides(1)(2).
1) Department of Biochemistry and Microbiology, Ghent University, Ghent, Belgium
(2) VIB-UGent Center for Inflammation Research, VIB, Ghent, Belgium
Plasmid reference: Verschueren et al., Nature 568 (2019), 571-575 [PMID: 30944476] [DOI: 10.1038/s41586-019-1095-5]
Restricted distribution: - VIB/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: 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 + ampicillin (100 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
Other culture collection numbers: -

Refer in your Materials and Methods:

pET-DUET-hACLY-A/B (LMBP 11131) is available at BCCM/GeneCorner. The plasmid was deposited by A. Dansercoer, Dr K. Verstraete and Prof. Dr S. Savvides and was published in Verschueren et al., 2019.

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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