Part:BBa_K4423021:Design

pTEF_MaSp_RPL2Att

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 1065
  Illegal PstI site found at 1260
  Illegal PstI site found at 1272
  Illegal PstI site found at 1419
  Illegal PstI site found at 1614
  Illegal PstI site found at 1626
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 1065
  Illegal PstI site found at 1260
  Illegal PstI site found at 1272
  Illegal PstI site found at 1419
  Illegal PstI site found at 1614
  Illegal PstI site found at 1626
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 819
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 1065
  Illegal PstI site found at 1260
  Illegal PstI site found at 1272
  Illegal PstI site found at 1419
  Illegal PstI site found at 1614
  Illegal PstI site found at 1626
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 1065
  Illegal PstI site found at 1260
  Illegal PstI site found at 1272
  Illegal PstI site found at 1419
  Illegal PstI site found at 1614
  Illegal PstI site found at 1626
  Illegal NgoMIV site found at 983
  Illegal AgeI site found at 2303
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

The main strain spider silk can not be synthesized in one piece due to a high number of repeats. Promoter and terminator are specific for K. phaffii

Source

Promoter and terminator sequences were provided by our PI Dr. Hans Marx, Department for Biotechnology of the University for Natural Ressources and Life Sciences, Vienna The sequence for the main strain spider silk protein was taken out of the paper Bin Liu, Tao Wang, Liyan Xiao, Guilan Zhang, Guangshen Li, Jingzhi Luo & Xiaobing Liu (2018) A directed self-assembly quasi-spider silk protein expressed in Pichia pastoris, Biotechnology & Biotechnological Equipment, 32:2, 451-461, DOI: 10.1080/13102818.2017.1327823