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GeneCorner plasmid details

Last data update: 22 October 2020 04:25 CEST

Plasmid name: BiFC3 (LMBP 10466)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p10466.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: B gene of the control of cell death locus of the Escherichia coli F plasmid (ccdB, lethal gene)
Aequorea victoria green fluorescent protein DNA (GFP); enhanced yellow-green fluorescent variant (EYFP), yeast codon-optimised enhanced monomeric version of Venus (yEmVenus), C-terminal fragment, N-terminal
Promoter: Escherichia coli lac operon promoter
Phage SP6 promoter
Escherichia coli β-lactamase promoter (amp)
Candida albicans sulfate adenylyltransferase promoter (CaMET3)
Ribosome
binding site:
-
Terminator: Candida albicans actin terminator (ACT1)
Selection marker: Neomycin (neo; G418)
Candida dubliniensis ARG4 (CdARG4); auxotrophic
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli; use a ccdB-resistant strain for propagation
Candida albicans; arg4(-), integrative
Parental clone: 2016-N
Further information: This plasmid was constructed by cloning the yEmVenus fragment (aa 173-238), preceeded by a start codon, into the BspEI/NheI sites of the 2016-N vector.
The plasmid is used for bimolecular fluorescence complementation in C. albicans, for studying protein-protein interactions.
The C-terminal fragment of the yEmVenus coding sequence is followed by a GGGGSGGGGS-linker, a multiple cloning site and a stop codon.
The plasmid contains an integration sequence, a ±850-bp Candida albicans genomic DNA fragment located between RXT3 and ORF19.3569 on chromosome 2. After restriction with NotI, the linearized vector will be targeted to this genomic site for integration (see Stynen et al. 2010).
The nucleotide sequence of the Candida albicans MET3 promoter and ACT1 terminator corresponds with the EMBL Nucleotide Sequence Database accession number CP017623.1.
The nucleotide sequence of the Candida dubliniensis ARG4 cDNA corresponds with the Genbank accession number NC_012866.1.
EMBL Accession number: CP017623.1, view at EMBL, GenBank, DDBJ
NC_012866.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 24/11/2017
Sequence detail:
Primers used to amplify the yEmVenus fragment (aa 173-238): 

                      173 174 175 176 177 178 179 180
B-7571: CCGTCCGGA.ATG.GAT.GGT.GGT.GTT.CAA.TTA.GCT.GAC
           BspEI  ***

                    238 237 236 235 234 233 232 231
B-7572: CCG.GCT.AGC.TTT.GTA.CAA.TTC.ATC.CAT.ACC.ATG.GG
            NheI        

***: start 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 Prof. Dr P. Van Dijck(1) (2). It was constructed by Dr E. Swinnen(1) (2).
(1) VIB-KU Leuven Center for Microbiology, VIB, Leuven, Belgium
(2) Laboratory of Molecular Cell Biology, Institute of Botany and Microbiology, KU Leuven, Leuven, Belgium
Plasmid reference: Subotic et al., G3 (Bethesda) 7 (2017), 3509-3520 [PMID: 28860184] [DOI: 10.1534/g3.117.300149]
Related plasmid reference: Stynen et al., Nucleic Acids Res. 38 (2010), e184 [PMID: 20719741] [DOI: 10.1093/nar/gkq725]
Restricted distribution: - BCCM MTA
- The depositor will be informed of the customer's identity upon release of a sample outside the depositor's department or outside the departments in which BCCM/GeneCorner is embedded, namely UGent-DBMB and VIB-IRC.
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 DH5α
Host reference: Focus 8 (1986), 9
Related host reference: Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051]
Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187]
Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791]
Rodriguez-Quinones et al., Focus 15 (1993), 110-112
Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660]
Cultivation medium: LB-Lennox + kanamycin (50 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
Other culture collection numbers: -

Refer in your Materials and Methods:

BiFC3 (LMBP 10466) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr P. Van Dijck and was published in Subotic et al., 2017.

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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