Table 1.

Dual clonase efficiencies: results of all dual clonase reactions using vectors designed to prevent misrecombination by virtue of the bacterial drug resistance marker positioned between the two tandem DEST cassettes

Vector	pENTR1	Size (kb)	pENTR2	Size (kb)	Correct Clones	% Correct
pDual DEST Shuttle (Cm)	DsRED2	0.82	EGFP	0.78	2/2	100
pDual DEST Shuttle (Cm)	Venus	0.78	AmCyan	0.75	3/4	75
pPB-Dual DEST (Kan)	EGFR-GFP	4.53	shP53-GFP	3.23	4/4	100
pPB-Dual DEST (Kan)	DsRED2	0.82	EGFP	0.78	4/4	100
pPB-Dual DEST (Kan)	EGFP	0.78	DsRED2	0.82	4/4	100
pPB-Dual DEST (Kan)	DsRED2	0.82	EGFP-IRES-Puro/TK	3.32	3/3	100
pPB-Dual DEST (Kan)	LargeT Antigen	2.81	NRASV12G	0.61	4/4	100
pPB-Dual DEST (Kan)	shP53-GFP	3.23	rtTA-IRES-Luc	3.22	4/4	100
pDual DEST Shuttle (Cm)	EGFP	0.78	Luciferase	1.91	3/3	100
pPB-Dual DEST (Kan)	EGFR	3.70	shP53-GFP	3.23	2/2	100
pDual DEST Shuttle (Cm)	EGFP	0.78	DsRED2	0.82	2/2	100
pPB-IDual DEST (Kan)	DsRED2	0.82	EGFP-IRES-Puro/TK	3.32	2/2	100
pDual DEST Shuttle (Cm)	EGFP	0.78	DsRED2	0.82	2/2	100
pDual DEST Shuttle (Spec)	mYFP	0.73	AmCyan	0.75	1/2	50
pIDual DEST Shuttle (Spec)	mYFP	0.73	AmCyan	0.75	2/2	100
pPB-IDual DEST (Kan)	DsRED2	0.82	EGFP-IRES-Puro/TK	3.32	2/2	100
pPB-Dual DEST (Kan)	shCREBBP-RFP	1.26	shPTEN-GFP	3.23	2/2	100
pPB-IDual DEST (Kan)	mKate	0.73	Puro-TK	1.99	2/2	100
pPB-Dual DEST (Kan)	EGFR-GFP	4.53	FOXR2-IRES-Puro/TK	3.56	2/2	100
pPB-IDual DEST (Kan)	mKATE	0.73	Puro-TK	1.99	2/2	100
pDual DEST Shuttle (Spec)	mKATE	0.73	Puro-TK	1.99	2/2	100
pPB-Dual DEST (Kan)	mKATE	0.73	Puro-TK	1.99	2/2	100
pPB-IDual DEST (Kan)	LargeT Antigen	2.81	NRASV12G-IRES-Luciferase	3.42	2/2	100
pDual DEST Shuttle (Spec)	eBFP2	0.73	mOrange2	0.90	2/2	100
pIDual DEST Shuttle (Spec)	eBFP2	0.73	mOrange2	0.90	2/2	100
pPB-IDual DEST (Kan)	M2-rtTA	0.77	Puro-TK	1.99	2/2	100
pDual DEST Shuttle (Cm)	EGFP	0.78	mKATE	0.73	2/2	100
pPB-Dual DEST (Kan)	mKATE	0.73	EGFP	0.78	2/2	100
pPB-IDual DEST (Kan)	mKATE	0.73	EGFP	0.78	2/2	100
pDual DEST Shuttle (Cm)	mKATE	0.73	EGFP	0.78	2/2	100
pDual DEST Shuttle (Cm)	EGFP	0.78	mKATE	0.73	2/2	100
pPB-IDual DEST (Kan)	EGFR-GFP	4.53	FOXR2-IRES-Puro/TK	3.56	2/2	100
pIDual DEST Shuttle (Cm)	shTrp53-GFP	3.63	NRASV12G	0.61	2/2	100
pIDual DEST Shuttle (Cm)	EGFP	0.78	Luciferase	1.91	2/2	100
pPB-IDual DEST (Kan)	Fah	1.30	Luciferase	1.91	1/1	100
pPB-IDual DEST (Kan)	LargeT Antigen	2.81	NRASV12G	0.61	2/2	100
pDual DEST Shuttle (Cm)	mYFP	0.73	AmCyan	0.75	2/2	100
pIDual DEST Shuttle (Cm)	mYFP	0.73	AmCyan	0.75	2/2	100
pPB-Dual DEST (Kan)	mYFP	0.73	mKATE	0.73	2/2	100
pPB-IDual DEST (Kan)	mYFP	0.73	mKATE	0.73	2/2	100
					91/93	98.10%