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For FAT10–Eos, FAT10–Eos–tail, FAT10 ΔUBL2 –Eos, FAT10 ΔUBL2 –Eos–tail, FAT10 ΔUBL1 –Eos and FAT10 ΔUBL1 –Eos–tail, the wild-type FAT10 vector (His–Smt3–FAT10) was linearized and PCR fragments of Eos–intein–CBD or Eos–tail–intein–CBD were inserted by HiFi assembly