Part:BBa_J119002

BsaI-GFP2-BsmBI

This is the second half of the GFP gene split for use in solving HPP (Hamiltonian Path Problems). It is the necessary complement to GFP-1 part name: BBa_J119001 It will function as a reporter if both genes are transcribed in proper order after re-assembly in E Coli. It is also intended to be mixed with other gene halves to attempt a multi-node HPP problem that will show correct completion if all reporters are present (resistance, fluorescence, etc.). All groupings must have both halves of a specific reporter gene to function properly.

Here is the design of the forward and reverse sequences minus the entire gene sequence.

GFP_2_For (36mer) GCAT GAATTC GGTCTC A ACCC TTGTTAATAGAATCG

(4nt, EcoRI , BsaI, a, 1234, 15mer of GFP_2)

GFP_2_Rev (158mer) ATGC CTGCAG CGTCTC A CCAC GTGCTCAGTATCTCTATCACTGATAGGGATGTCAATCTCTATCACTGATAGGGA TTGC

(4nt , PstI , BsmBI , a,1234, pTet Reverse Complement , 4nt)

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 1
  Illegal PstI site found at 452
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 1
  Illegal PstI site found at 452
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 1
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 1
  Illegal PstI site found at 452
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 1
  Illegal PstI site found at 452
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 7
  Illegal BsaI.rc site found at 306