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GeneCorner plasmid details

Last data update: 27 October 2020 04:27 CET

Plasmid name: BiFC2 (LMBP 10465)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p10465.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Aequorea victoria green fluorescent protein DNA (GFP); enhanced yellow-green fluorescent variant (EYFP), yeast codon-optimised enhanced monomeric version of Venus (yEmVenus), N-terminal fragment, C-terminal
B gene of the control of cell death locus of the Escherichia coli F plasmid (ccdB, lethal gene)
Promoter: Candida albicans sulfate adenylyltransferase promoter (CaMET3)
Escherichia coli lac operon promoter
Phage SP6 promoter
Phage T7 gene 10 promoter (T7g10)
Escherichia coli β-lactamase promoter (amp)
Ribosome
binding site:
-
Terminator: Candida albicans actin terminator (ACT1)
Selection marker: Neomycin (neo; G418)
Candida maltosa LEU2 (CmLEU2); auxotrophic
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli; use a ccdB-resistant strain for propagation
Candida albicans; leu2(-), integrative
Parental clone: 2015-C
Further information: This plasmid was constructed by cloning the yEmVenus fragment (aa 1-172) followed by a stop codon, into the NheI/MluI sites of the 2015-C vector.
The plasmid is used for bimolecular fluorescence complementation in C. albicans, for studying protein-protein interactions.
The N-terminal fragment of the yEmVenus coding sequence is preceeded by a start codon, a multiple cloning site and a GGGGSGGGGS-linker.
The plasmid contains an integration sequence, a ±1100-bp Candida albicans genomic DNA fragment located between XOG1 and HOL1 on chromosome 1. 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.
EMBL Accession number: CP017623.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 17/11/2017
Sequence detail:
Primers used to amplify the yEmVenus fragment (aa 1-172): 

                   1   2   3   4   5   6   7   8   9
B-7569: CCGGCTAGC.ATG.TCT.AAA.GGT.GAA.GAA.TTA.TTC.ACT.GG
           NheI   ***

                      172 171 170 169 168 167 166 165 164 163
B-7570: CCGACGCGT.TTA.TTC.AAT.GTT.GTG.TCT.AAT.TTT.GAA.GTT.AGC
           MluI   +++

***: 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 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:

BiFC2 (LMBP 10465) 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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