Part:BBa_K1138002

pAW50-linker-mcherry(+RBS) (ugaiGEM12)

This part is similar to part BBa_K1138001 with the difference that an RBS sequence is inserted between the XbaI and NsiI restriction sites. This part can serve as a control while doing experiments utilizing part BBa_K1138001. This plasmid consists of the mcherry gene (encoding red fluorescence protein) inserted in the backbone of pAW50 vector. EcoRI-NotI-XbaI-RBS-NsiI restriction sites have been introduced followed by a linker sequence followed by the mcherry sequence. The use of these sites will allow users to introduce any desired gene of their choice to be fused with the linker which is already fused with the mcherry sequence. This way any protein can be qualitatively studied in Methanogens via red fluorescence. Note:- the users will have to introduce the RBS sequence and start codon, delete the terminator sequence of their gene while cloning.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 199
  Illegal EcoRI site found at 205
  Illegal XbaI site found at 220
  Illegal SpeI site found at 975
  Illegal PstI site found at 615
  Illegal PstI site found at 982
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 199
  Illegal EcoRI site found at 205
  Illegal SpeI site found at 975
  Illegal PstI site found at 615
  Illegal PstI site found at 982
  Illegal NotI site found at 211
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 199
  Illegal EcoRI site found at 205
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal prefix found at 205
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 199
  Illegal SpeI site found at 975
  Illegal PstI site found at 615
  Illegal PstI site found at 982
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal prefix found at 205
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 199
  Illegal XbaI site found at 220
  Illegal SpeI site found at 975
  Illegal PstI site found at 615
  Illegal PstI site found at 982
  Illegal AgeI site found at 993
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal BsaI site found at 3151
  Illegal BsaI.rc site found at 1534
  Illegal BsaI.rc site found at 1863
  Illegal SapI site found at 4815