Versatile approach for functional analysis of human proteins and efficient stable cell line generation using FLP-mediated recombination system.

Szczesny RJ, Kowalska K, Klosowska-Kosicka K, Chlebowski A, Owczarek EP, Warkocki Z, Kulinski TM, Adamska D, Affek K, Jedroszkowiak A, Kotrys AV, Tomecki R, Krawczyk PS, Borowski LS, Dziembowski A
PLoS One. 2018 Mar 28;13(3):e0194887. doi: 10.1371/journal.pone.0194887. eCollection 2018. PubMed Article

Plasmids from Article

ID	Plasmid	Purpose
105767	pKK-NoTag	Control vector for pKK series (encodes only SLIC arms (TEV-L and TEV-R)); expression without tag. Useful to design other vectors; any tag can be added.
105768	pKK-FLAG-TEV	Expression of your protein of interest in fusion with FLAG at the N-terminus. The tag is cleavable by TEV protease or enterokinase.
105769	pKK-HA-TEV	Expression of your protein of interest in fusion with HA at the N-terminus. The tag is cleavable by TEV protease.
105770	pKK-FLAG-BirA-TEV	Expression of your protein of interest in fusion with FLAG-BirA. FLAG-tagged at the N-terminus (cleavable by TEV). Search for protein partners using the BioID (PMID: 15707685, 22412018).
105771	pKK-MBP-TEV	Expression of your protein of interest in fusion with MBP at the N-terminus (cleavable by TEV). The maltose binding protein tag allows convenient purification of proteins (PMID: 22442034, 19935684).
105772	pKK-mTagBFP-TEV	Expression of your protein of interest in fusion with blue fluorescent protein at the N-terminus (cleavable by TEV) (PMID: 18940671, 25378625). Useful for multi-channel live cell imaging.
105773	pKK-mCerulean-TEV	Expression of your protein of interest in fusion with cyan fluorescent protein at the N-terminus (cleavable by TEV). Useful for FRET experiments (PMID: 14990965, 17040988, 22264545).
105774	pKK-mVenus-TEV	Expression of your protein of interest in fusion with yellow fluorescent protein at the N-terminus (cleavable by TEV). Useful for FRET experiments (PMID: 11753368, 17040988, 22264545).
105775	pKK-mAmber-TEV	Expression of your protein of interest in fusion with non-absorbing, non-emitting Venus mutant at the N-terminus (cleavable by TEV). Negative control for FRET measurements (PMID: 17040988, 22264545).
105776	pKK-EGFP-TEV	Expression of your protein of interest in fusion with green fluorescent protein at the N-terminus (cleavable by TEV), (PMID: 8885998, 8805248). EGFP is not truly monomeric (PMID: 11988576, 22869113)
105777	pKK-mEGFP-TEV	Expression of your protein of interest in fusion with green fluorescent protein at the N-terminus (cleavable by TEV). mEGFP is an EGFP A206K mutant with reduced dimerization (PMID: 11988576,22869113).
105778	pKK-mClover3-TEV	Expression of your protein of interest in fusion with green fluorescent protein at the N-terminus (cleavable by TEV). mClover3 is a brighter derivative of mEGFP (PMID: 26879144).
105779	pKK-mCherry-TEV	Expression of your protein of interest in fusion with red fluorescent protein at the N-terminus (cleavable by TEV), (PMID: 15558047). mCherry descends from the first truly monomeric red FP mRFP1.
105780	pKK-mRuby3-TEV	Expression of your protein of interest in fusion with red fluorescent protein at the N-terminus (cleavable by TEV), (PMID: 26879144). mRuby3 is brighter than mCherry and has shifted ex and em spectra.
105781	pKK-mCardinal-TEV	Expression of your protein of interest in fusion with red fluorescent protein at the N-terminus (cleavable by TEV), (PMID: 24633408). mCardinal is suitable for deep-tissue imaging.
105782	pKK-CyOFP1-TEV	Expression of your protein of interest in fusion with orange fluorescent protein at the N-terminus (cleavable by TEV) (PMID: 27240196). CyOFP1 is useful for multi-channel imaging.
105783	pKK-TEV-FLAG	Expression of your protein of interest in fusion with FLAG at the C-terminus. The tag is cleavable by TEV protease.
105784	pKK-TEV-3XFLAG	Expression of your protein of interest in fusion with 3xFLAG at the C-terminus. The tag is cleavable by TEV protease.
105785	pKK-TEV-HA	Expression of your protein of interest in fusion with HA at the C-terminus. The tag is cleavable by TEV protease.
105786	pKK-TEV-BirA-FLAG	Expression of your protein of interest in fusion with BirA-FLAG at the C-terminus (cleavable by TEV). Search for protein partners using the BioID (PMID: 15707685, 22412018).
105787	pKK-TEV-MBP	Expression of your protein of interest in fusion with MBP at the C-terminus (cleavable by TEV). The maltose binding protein tag allows convenient purification of proteins (PMID: 22442034, 19935684).
105788	pKK-TEV-ProteinA	Expression of your protein of interest in fusion with two IgG binding domains of Staphylococcus aureus protein A at the C-terminus (cleavable by TEV) (PMID: 3507693, 10504710). Protein purification.
105789	pKK-TEV-mTagBFP	Expression of your protein of interest in fusion with blue fluorescent protein at the C-terminus (cleavable by TEV) (PMID: 18940671, 25378625). Useful for multi-channel live cell imaging.
105790	pKK-TEV-mCerulean	Expression of your protein of interest in fusion with cyan fluorescent protein at the C-terminus (cleavable by TEV). Useful for FRET experiments (PMID: 14990965, 17040988, 22264545).
105791	pKK-TEV-mVenus	Expression of your protein of interest in fusion with yellow fluorescent protein at the C-terminus (cleavable by TEV). Useful for FRET experiments (PMID: 11753368, 17040988, 22264545).
105792	pKK-TEV-mAmber	Expression of your protein of interest in fusion with non-absorbing, non-emitting Venus mutant at the C-terminus (cleavable by TEV). Negative control for FRET measurements (PMID: 17040988, 22264545).
105793	pKK-TEV-EGFP	Expression of your protein of interest in fusion with green fluorescent protein at the C-terminus (cleavable by TEV), (PMID: 8885998, 8805248). EGFP is not truly monomeric (PMID: 11988576, 22869113)
105794	pKK-TEV-mEGFP	Expression of your protein of interest in fusion with green fluorescent protein at the C-terminus (cleavable by TEV). mEGFP is an EGFP A206K mutant with reduced dimerization (PMID: 11988576,22869113).
105795	pKK-TEV-mClover3	Expression of your protein of interest in fusion with green fluorescent protein at the C-terminus (cleavable by TEV). mClover3 is a brighter derivative of mEGFP (PMID: 26879144).
105796	pKK-TEV-mCherry	Expression of your protein of interest in fusion with red fluorescent protein at the C-terminus (cleavable by TEV), (PMID: 15558047). mCherry descends from the first truly monomeric red FP mRFP1.
105797	pKK-TEV-mRuby3	Expression of your protein of interest in fusion with red fluorescent protein at the C-terminus (cleavable by TEV) (PMID: 26879144). mRuby3 is brighter than mCherry and has shifted ex and em spectra.
105798	pKK-TEV-mCardinal	Expression of your protein of interest in fusion with red fluorescent protein at the C-terminus (cleavable by TEV), (PMID: 24633408). mCardinal is suitable for deep-tissue imaging.
105799	pKK-TEV-CyOFP1	Expression of your protein of interest in fusion with orange fluorescent protein at the C-terminus (cleavable by TEV) (PMID: 27240196). CyOFP1 is useful for multi-channel imaging.
105800	pKK-BI16-TEV-mCherry	Concomitant expression of two proteins. One protein is expressed with mCherry fusion tag (at the C-terminus), cleavable by TEV protease; second tag introduced by user.
105801	pKK-BI16-FLAG-3C-ORF1_mClover3-TEV-ORF2	Concomitant expression of two proteins. One protein expressed with FLAG, cleavable by 3C or enterokinase; second protein expressed with mClover3, cleavable by TEV protease; tags positions: N-termini.
105802	pKK-BI16-ORF1-3C-mRuby3_ORF2-TEV-mClover3	Concomitant expression of two proteins. One protein expressed with mRuby3, cleavable by 3C; second protein expressed with mClover3, cleavable by TEV protease; tags positions: C-termini.
105803	pKK-BI16-ORF1-3C-FLAG_ORF2-TEV-mClover3	Concomitant expression of two proteins. One protein expressed with FLAG, cleavable by 3C protease; second protein expressed with mClover3, cleavable by TEV protease; tags positions: C-termini.
105804	pKK-BiFC-Venus	Concomitant expression of two proteins in fusion with fragments of Venus protein. Protein interactions studies by bimolecular fluorescence complementation (BiFC), including BiFC-FRET assays.
105805	pKK-FRET-ORF1-3C-Cerulean_ORF2-TEV-Venus	Concomitant expression of two proteins. One protein expressed with Cerulean, cleavable by 3C; second protein expressed with Venus, cleavable by TEV. Useful for protein interaction analysis by FRET.
105806	pKK-FRET-ORF1-3C-Cerulean_ORF2-TEV-Amber	Concomitant expression of two proteins. One protein expressed with Cerulean, cleavable by 3C; second protein expressed with Amber, cleavable by TEV. Control for protein interaction analysis by FRET.
105807	pKK-RNAi-nucEGFPmiR-FLAG-TEV	Functional studies using RNAi; rescue experiments; substituting protein for its different form. miRNA cassette with nuclear EGFP expression marker; your protein of interest with N-terminal FLAG tag.
105808	pKK-RNAi-nucEGFPmiR-mCherry-TEV	Functional studies using RNAi; rescue experiments; substituting protein for its different form. miRNA cassette with nuclear EGFP expression marker; your protein of interest with N-terminal mCherry.
105809	pKK-RNAi-nucCHERRYmiR-FLAG-TEV	Functional studies using RNAi; rescue experiments; substituting protein for its different form. miRNA cassette with nuclear mCherry expression marker; your protein of interest with N-terminal FLAG.
105810	pKK-RNAi-nucCHERRYmiR-EGFP-TEV	Functional studies using RNAi; rescue experiments; substituting protein for its different form. miRNA cassette with nuclear mCherry expression marker; your protein of interest with N-terminal EGFP.
105811	pKK-RNAi-nucEGFPmiR-TEV-FLAG	Functional studies using RNAi; rescue experiments; substituting protein for its different form. miRNA cassette with nuclear EGFP expression marker; your protein of interest with C-terminal FLAG.
105812	pKK-RNAi-nucEGFPmiR-TEV-mCherry	Functional studies using RNAi; rescue experiments; substituting protein for its different form. miRNA cassette with nuclear EGFP expression marker; your protein of interest with C-terminal mCherry.
105813	pKK-RNAi-nucCHERRYmiR-TEV-FLAG	Functional studies using RNAi; rescue experiments; substituting protein for its different form. miRNA cassette with nuclear mCherry expression marker; your protein of interest with C-terminal FLAG.
105814	pKK-RNAi-nucCHERRYmiR-TEV-EGFP	Functional studies using RNAi; rescue experiments; substituting protein for its different form. miRNA cassette with nuclear mCherry expression marker; your protein of interest with C-terminal EGFP.