Last data update: 24 January 2024 16:39 CET
Plasmid name: pCAGGS-E-CYLD (LMBP 6613)
| New search | Print data sheet |
| Price category: | Cat. 3 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p6613.gb
(View with Genome Compiler) p6613.txt p6613.pdf |
| Sequence analysis results Genecorner: |
NGS: c09-gc-dec2018.fasta |
| Cloned DNA: |
Human CYLD lysine 63 deubiquitinase cDNA (CYLD, CYLD1, USPL2, GeneID 1540) 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: | pCD-F-CYLD; pCAGGSEhA20 |
| Further information: | The plasmid was constructed by inserting the NotI/XhoI PCR fragment, amplified from pCD-F-CYLD and containing the human CYLD coding sequence, into the NotI/XhoI opened pCAGGSEhA20 vector, replacing the human A20 coding sequence. The plasmid is useful for highly efficient expression of human CYLD 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 this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT726834.1. The nucleotide sequence of the human CYLD cDNA corresponds with the EMBL Nucleotide Sequence database accession number AJ250014.1. |
| EMBL Accession number: | AJ250014.1, view at EMBL, GenBank, DDBJ LT726834.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 19/04/2011 |
| Sequence detail: | Primers used to amplify the human CYLD coding sequence:
Forward: 5' TCGTGCGGCCGCA.ATG.AGT.TCA.GGC.TTA.TGG.AGC
NotI ***
Reverse: 5' GGCCTCGAGGTC.TTA.TTT.GTA.CAA.ACT.CAT.TGT.TGG.A
XhoI +++
***: start codon.
+++: termination codon.
Punctuation indicates reading frame. |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: NotI/XhoI, PvuI, SalI and XmnI. This plasmid has also been fully sequenced but the NGS sequence data still need to be implemented. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Dr 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: | Staal et al., EMBO J. 30 (2011), 1742-1752 [PMID: 21448133] [DOI: 10.1038/emboj.2011.85] |
| Related plasmid reference: | Staal et al., bioRxiv (2016), 1-42 [DOI: 10.1101/046789] Lork et al., Front. Cell Dev. Biol. 6 (2018), 40 [PMID: 29755980] [DOI: 10.3389/fcell.2018.00040] Afonina et al., EMBO Rep. 17 (2016), 914-927 [PMID: 27113748] [DOI: 10.15252/embr.201642109] |
| 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: | - |
Refer in your Materials and Methods: |
pCAGGS-E-CYLD (LMBP 6613) is available at BCCM/GeneCorner. This plasmid was deposited by Dr J. Staal and Prof. Dr R. Beyaert and was published in Staal et al., 2011. |
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.