Last data update: 24 January 2024 16:39 CET
Plasmid name: pCD-F-hCYLD-N (LMBP 6113)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p6113.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human CYLD lysine 63 deubiquitinase cDNA (CYLD, CYLD1, USPL2, GeneID 1540); N-terminal fragment FLAG epitope tag; N-terminal |
| Promoter: | Human cytomegalovirus immediate early promoter (CMV-IE) and enhancer Phage T7 gene 10 promoter (T7g10) Simian virus 40 early promoter (SV40 early) Escherichia coli lac operon promoter |
| Ribosome binding site: |
- |
| Terminator: | Bovine growth hormone polyadenylation signal (BGH polyA) Simian virus 40 polyadenylation signal (SV40 polyA) |
| Selection marker: | Ampicillin (amp) Neomycin (neo; G418) |
| Replicon: | Escherichia coli plasmid pMB1 origin Phage f1 origin Simian virus 40 bidirectional origin (SV40) |
| Host range: | Escherichia coli Mammalian cells; SV40 permissive cells |
| Parental clone: | pCD-F-MLTp20; pCDNA3-Flag-CYLD |
| Further information: | The plasmid was created by PCR amplifying the N-terminal fragment of the hCYLD cDNA from pCDNA3-Flag-CYLD and cloning the fragment into the KpnI-ApaI opened pCD-F-MLTp20 vector. The plasmid contains an N-terminal fragment of hCYLD, consisting of amino acids 1 up to 324. The parental clone of this vector has been shown to provide resistance to kanamycin, although growth was reduced compared to growth on medium containing ampicillin. The nucleotide sequence of the wild type human CYLD cDNA corresponds with the EMBL Nucleotide Sequence database accession numberAJ250014.1. Other name of the plasmid is pCD-F-CYLD-N. |
| EMBL Accession number: | AJ250014.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 02/04/2009 |
| Sequence detail: | Primers used to amplify the N-terminal fragment of the CYLD gene:
Forward: 5' ATGGTACCA.ATG.AGT.TCA.GGC.TTA.TGG.AGC
KpnI ***
Reverse: 5' TAGGGGCCC.CTA.ATT.TCC.AGG.GTC.TGA.GGT.AG
ApaI +++
***: start codon.
+++: termination codon.
Punctuation indicates reading frame. |
| Authenticity test: | The plasmid still needs to be subjected to the authenticity test. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by J. Staal(1) (2) and 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 - Possible extra restrictions defined by the depositor, based on the description of the enduser's research |
| 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: | - |
Refer in your Materials and Methods: |
pCD-F-hCYLD-N (LMBP 6113) is available at BCCM/GeneCorner. This plasmid was deposited by J. Staal and 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.