Table II

Plasmid Construction

Plasmid ConstructionReference
pRS313 Backbone
 pSW363NUP145 locus lacking base pairs 2,191–2,850 (the GLFG region)Emtage et al., 1997
pRS315 Backbone
 pSW503Entire KAP95 locus in BamHI in pRS315This study
 pSW509Entire kap95-L63A locus in BamHI in pRS315This study*
 pSW696Entire kap95-ΔNES locus in BamHI in pRS315This study*
pRS316 Backbone
 pSW131Entire NUP116 locus in SalI/Xbal of pRS316Wente and Blobel, 1994
 pSW132Entire NUP100 locus is SalI/Xbal of pRS316Wente and Blobel, 1994
 pSW190Entire NUP145 locus in BamHI/SalI of pRS316Emtage et al., 1997
 pSW271Entire KAP95 locus in BamHI of pRS316Iovine et al., 1995
 pSW461Insertion of five tandem IgG-binding domains into NsiI site at base pair 990 of NUP116 locusThis study*
pRS304 Backbone
 pSW251NUP116 locus lacking base pairs 1,260–2,873 (the GLFG region)This study
pRS305 Backbone
 pSW263NUP100 locus lacking base pair 890–2,657 (the GLFG region)This study
 pSW645Insertion of five tandem IgG-binding domains into NsiI site at base pair 1,253 of NUP100 locusThis study
pRS306 Backbone
 pSW280NUP145 locus lacking base pairs 2,191–2,850 (the GLFG region)
pCH436 Backbone
 pSW511kap95-L63A coding region in BamHIThis study
pCH358 Backbone
 pSW368SRP1 coding region in BamHIMurphy et al., 1995
 pSW392Fragment from base pair 890–2,657 (GLFG domain) of NUP100 locus in BamHIMurphy and Wente, 1996
 pSW608Fragment from base pair 1,294–2,446 of (FXFG domain) NUP1 locus in BamHIThis study
pGEX-3X Backbone
 pSW304Fragment from base pair 1,260–2,873 (GLFG domain) of NUP116 locus in BamHIThis study
 pSW568Fragment from base pair 3,383–3,641 (encoding amino acids 1–77) of KAP95 locus in BamHI/EcoRIThis study
 pSW570Fragment from base pair 3,383–3,641 (encoding amino acids 1–77) of kap95-ΔNES locus in BamHI/EcoRIThis study
pGEX-2T Backbone
 pPS826GSP1 coding regionCorbett et al., 1995
pQE-32 Backbone
 pSW329KAP95 coding region in BamHIThis study
 pSW571kap95-L63A coding region in BamHIThis study
  • Vector backbone references for pRS313, pRS315, pRS316, pRS304, pRS305, and pRS306 (Sikorski and Hieter, 1989); pCH436 for LexABD fusion proteins, and pCH358 for GAL4AD fusion proteins (generous gifts of C. Hardy); pQE-32 (Qiagen); pGEX-3X (Pharmacia Fine Chemicals).  

  • * See Materials and Methods section for a more detailed description.