Table I.

Plasmids used in this study

Plasmid name (ASP) Construct 
451 pGEX-6P-1 (GE Healthcare) containing a modified MCS for bacterial expression of GST-tagged proteins 
1,633 GST–xM18BP1-2161–415 
1,980 GST–xM18BP1-1161–375 
1,639 GST-xHJURP 
1,938 6His-xHJURP42–194 
447 pCS2+ containing a modified MCS for IVT of the protein in rabbit reticulocyte lysate or in vitro transcription of RNA 
459 pCS2+ containing 6×myc sequence upstream of the MCS for IVT of the protein in rabbit reticulocyte lysate or in vitro transcription of RNA 
1,730 pCS2+ containing 3×FLAG sequence upstream of the MCS for IVT of the protein in rabbit reticulocyte lysate or in vitro transcription of RNA 
1,853 pCS2+ containing 3×FLAG sequence downstream of the MCS for IVT of protein in rabbit reticulocyte lysate or in vitro transcription of RNA 
1,592 p447-xHJURP for production of xHJURP RNA 
1,815 p447-xHJURP (codon optimized) for production of the xHJURP IVT protein 
1,970 p447-hHJURP (codon optimized) 
1,968 p1853-xHJURP (codon optimized) 
1,969 p1853-hHJURP (codon optimized) 
559 p459–xCENP-A 
1,819 p1730–xCENP-A 
1,870 p1853–xM18BP1-1 
1,871 p1853–xM18BP1-2 
867 p459–xCENP-C (codon optimized) 
1,330 p459–xCENP-C amino acids 1–380 (codon optimized) 
1,925 p459–xCENP-C amino acids 200–579 (codon optimized) 
1,928 p459–xCENP-C amino acids 381–711 (codon optimized) 
1,926 p459–xCENP-C amino acids 580–949 (codon optimized) 
1,920 p459–xCENP-C amino acids 712–1,190 (codon optimized) 
1,927 p459–xCENP-C amino acids 950–1,291 (codon optimized) 
1,150 p459–xCENP-C amino acids 1,191–1,400 (codon optimized) 
1,943 p459–xCENP-C amino acids 1,204–1,400 (codon optimized) 
1,935 p459–xCENP-C amino acids 1,229–1,400 (codon optimized) 
1,921 p459–xCENP-C amino acids 1,191–1,350 (codon optimized) 
1,936 p459–xCENP-C amino acids 1,191–1,310 (codon optimized) 
866 p447–xCENP-C (codon optimized) 
1,350 p447–xM18BP1-1 
1,351 p447–xM18BP1-2 
844 p447–hCENP-C 
1,828 p447-hM18BP1 (codon optimized) 
572 p447–mCherry-tubulin 
1,981 p459-dTm2 (Drosophila melanogaster tropomyosin 2) 
Plasmid name (ASP) Construct 
451 pGEX-6P-1 (GE Healthcare) containing a modified MCS for bacterial expression of GST-tagged proteins 
1,633 GST–xM18BP1-2161–415 
1,980 GST–xM18BP1-1161–375 
1,639 GST-xHJURP 
1,938 6His-xHJURP42–194 
447 pCS2+ containing a modified MCS for IVT of the protein in rabbit reticulocyte lysate or in vitro transcription of RNA 
459 pCS2+ containing 6×myc sequence upstream of the MCS for IVT of the protein in rabbit reticulocyte lysate or in vitro transcription of RNA 
1,730 pCS2+ containing 3×FLAG sequence upstream of the MCS for IVT of the protein in rabbit reticulocyte lysate or in vitro transcription of RNA 
1,853 pCS2+ containing 3×FLAG sequence downstream of the MCS for IVT of protein in rabbit reticulocyte lysate or in vitro transcription of RNA 
1,592 p447-xHJURP for production of xHJURP RNA 
1,815 p447-xHJURP (codon optimized) for production of the xHJURP IVT protein 
1,970 p447-hHJURP (codon optimized) 
1,968 p1853-xHJURP (codon optimized) 
1,969 p1853-hHJURP (codon optimized) 
559 p459–xCENP-A 
1,819 p1730–xCENP-A 
1,870 p1853–xM18BP1-1 
1,871 p1853–xM18BP1-2 
867 p459–xCENP-C (codon optimized) 
1,330 p459–xCENP-C amino acids 1–380 (codon optimized) 
1,925 p459–xCENP-C amino acids 200–579 (codon optimized) 
1,928 p459–xCENP-C amino acids 381–711 (codon optimized) 
1,926 p459–xCENP-C amino acids 580–949 (codon optimized) 
1,920 p459–xCENP-C amino acids 712–1,190 (codon optimized) 
1,927 p459–xCENP-C amino acids 950–1,291 (codon optimized) 
1,150 p459–xCENP-C amino acids 1,191–1,400 (codon optimized) 
1,943 p459–xCENP-C amino acids 1,204–1,400 (codon optimized) 
1,935 p459–xCENP-C amino acids 1,229–1,400 (codon optimized) 
1,921 p459–xCENP-C amino acids 1,191–1,350 (codon optimized) 
1,936 p459–xCENP-C amino acids 1,191–1,310 (codon optimized) 
866 p447–xCENP-C (codon optimized) 
1,350 p447–xM18BP1-1 
1,351 p447–xM18BP1-2 
844 p447–hCENP-C 
1,828 p447-hM18BP1 (codon optimized) 
572 p447–mCherry-tubulin 
1,981 p459-dTm2 (Drosophila melanogaster tropomyosin 2) 

ASP, A.F. Straight plasmid.

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