GREAT AT SMALL THINGS

0

GeneCorner plasmid details

Last data update: 21 October 2020 04:26 CEST

Plasmid name: pCAGGS-E-CYLD (LMBP 6613)

Add to cart

New search

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: Afonina et al., EMBO Rep. 17 (2016), 914-927 [PMID: 27113748] [DOI: 10.15252/embr.201642109]
Lork et al., Front Cell Dev Biol 6 (2018), 40 [PMID: 29755980] [DOI: 10.3389/fcell.2018.00040]
Staal et al., bioRxiv (2016) [DOI: 10.1101/046789]
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.

New search