GeneCorner plasmid details

Last data update: 30 July 2021 10:22 CEST

Plasmid name: pCAGGS-E-hOPTN (LMBP 6026)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence:
analysis results


Cloned DNA: Human optineurin cDNA (OPTN, FIP2, GLC1E, HYPL, HIP7, GeneID 10133)
E-tag; N-terminal
Promoter: Chicken β-actin/rabbit β-globin hybrid promoter (AG)
Human cytomegalovirus immediate early promoter (CMV-IE); enhancer only
Escherichia coli lac operon promoter
binding site:
Terminator: Rabbit β-globin polyadenylation signal (β-globin polyA)
Simian virus 40 polyadenylation signal (SV40 polyA)
Selection marker: Ampicillin (amp)
Replicon: Escherichia coli plasmid pMB1 origin
Simian virus 40 bidirectional origin (SV40)
Host range: Escherichia coli
Mammalian cells; SV40 permissive cells
Parental clone: pCAGGS-E-hABIN1
Further information: The plasmid was constructed by inserting a 1742 bp NotI-XhoI PCR fragment, containing the human OPTN coding sequence, into the NotI-XhoI opened pCAGGS-E-hABIN1 vector.
pCAGGS-E-hOPTN is useful for highly efficient expression of human OPTN, fused in phase to the N-terminal E-tag, under the control of the AG promoter and the human CMV-IE enhancer in various mammalian cells.
When cloning a fragment downstream from the lac promoter it may be advisable to use lacI(q) strains in order to prevent fortuitous expression of a possibly noxious polypeptide.
The nucleotide sequence of the human OPTN cDNA corresponds with the EMBL Nucleotide Sequence Database accession number NM_001008213.1, except for a silent mutation at amino acid position 34.
Other names of the plasmid are pCAGGS-E-OPTN-FL and pCAGGS-E-hFIP2.
EMBL Accession number: NM_001008213.1, view at GenBank
Latest sequence update: 21/12/2005
Sequence detail:
Primers used to amplify the human OPTN cDNA:

                         BamHI   NotI      ***

reverse primer: 5' ... GCCGCTCGAG.TTA.AAT.GAT.GCA.ATC.CAT.CAC.GTG.AAT.CTG ... 3'
                           XhoI   +++

                                     32      34      36
WT according to NM_001008213: 5' ... CTG.GAC.ACG.TTT.ACC ... 3'
                                     Leu Asp Thr Phe Thr

                                     32      34      36
silent mutant:                5' ... CTG.GAC.ACA.TTT.ACC ... 3'
                                     Leu Asp Thr Phe Thr
***: start codon.
+++: termination codon.
^: mutated nucleotide.
Punctuation indicates reading frame.
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BstXI/EcoRI, HincII/NcoI, HindIII and NotI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Prof. Dr R. Beyaert(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: -
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 MC1061
Host reference: Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493]
Related host reference: Brigé et al., Biochem. J. 394 (2006), 335-344 [PMID: 16293111]
Cultivation medium: LB-Lennox + ampicillin (100 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
Cultivation remark: -
Other culture collection numbers: LMBP 05125

Refer in your Materials and Methods:

pCAGGS-E-hOPTN (LMBP 6026) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Beyaert .

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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