Modification of the Creator recombination system for proteomics applications – improved expression by addition of splice sites

Table 1 donor vectors constructs

*Vector ID*	*Vector Name*	*Potential Splicing Events*	*Description*
V1	pDNR Dual	Splice for 3' tags	Designed for Fusions at both N- and C- termini (Clontech)
V7	pDNR-MCS	Splice for 3' tags	Alternate MCS that adds AscI and PacI sites to the MCS of V1
V37	pDNR MCS SA	Splice for 5' and 3' tags	A splice acceptor introduced into V7 before the AscI site
V308	pDNR MCS lacZ alpha	Splice for 3' tags	LacZ alpha inserted between AscI and PacI sites of V7
V309	pDNR MCS SA lacZ alpha	Splice for 5' and 3' tags	LacZ alpha inserted between AscI and PacI sites of V37
V624	pDNR MCS BE	Splice for 3' tags	Alternate MCS with BamHI, SalI, SmaI, HindIII, EcoRI sites
V677	pDNR MCS BE lacZ alpha	Splice for 3' tags	LacZ alpha inserted between SalI and HindII sites of V624
V678	pDNR MCS SA ES	Splice for 5' and 3' tags	Alternate MCS Splice Acceptor with AscI, EcoRI, NotI, BamHI, SalI, PacI sites.
V795	pDNR MCS ES	Splice for 3' tags	Alternate MCS with AscI, EcoRI, NotI, BamHI, SalI, PacI sites
V954	pDNR MCS SA no SA CmR	Splice for 5' and 3' tags	Silent mutations introduced in the CmR ORF of V309 to remove potential splice acceptor

*Abbreviations used: SA – splice acceptor, MCS – multiple cloning site, CmR – chloramphenicol resistance, BE – BamHI/EcoRI, ES – EcoRI/SalI

ISSN: 1472-6750