Last data update: 24 January 2024 16:39 CET
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: | p6026.gb |
Sequence analysis results Genecorner: |
- |
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 |
Ribosome 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: forward primer: 5' ... CGGGATCCTAGCGGCCGCC.ATG.TCC.CAT.CAA.CCT.CTC.AGC.TGC.CTC ... 3' 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.