Part:BBa_K4364000

mRFP1-based cassette for TA cloning with dual T7 promotors

This part can be used to create a T-vector that is compatible with various TA cloning protocols. In order to do this, it should be cloned in a suitable pSB vector, this plasmid must be extracted and then treated with the restriction endonuclease AhdI. It will produce a linearized vector with single T overhangs. Moreover, the TA-cassette is integrated into the reading frame of mRFP1 under the control of the lac promoter. The empty vector produces notable red-colored colonies. If the TA cloning is successful the positive transformants will be white.

Please be careful if you transfer this part to pSB_A_ plasmid backbones - the beta-lactamase encoded by these vectors contains an AhdI site that should be destroyed first.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NotI site found at 199
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 207
  Illegal BamHI site found at 316
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 906
  Illegal AgeI site found at 1018
  
• 1000
  COMPATIBLE WITH RFC[1000]