Part:pSB1T3:Design

High copy BioBrick assembly plasmid

Assembly Compatibility:

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

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 2440
  Illegal NheI site found at 1268
  Illegal SpeI site found at 2
  Illegal PstI site found at 16
  Illegal NotI site found at 9
  Illegal NotI site found at 2446
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 2440
  Illegal BamHI site found at 1414
  Illegal XhoI site found at 1033
  Illegal XhoI site found at 2316
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal prefix found at 2440
  Illegal suffix found at 2
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal prefix found at 2440
  Plasmid lacks a suffix.
  Illegal XbaI site found at 2455
  Illegal SpeI site found at 2
  Illegal PstI site found at 16
  Illegal NgoMIV site found at 1440
  Illegal NgoMIV site found at 1808
  Illegal NgoMIV site found at 1968
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  

Design Notes

The ampicillin resistance marker needed to be eliminated.

In conversation with Austin Che, I gather this plasmid was made by taking a plasmid that was probably pSB1A3 and replacing the antibiotic resistance marker and sequence between two XhoI sites with a PCR product that encoded the Tetracycline resistance marker and could be digested with XhoI. This was done in a non-directional manner. Sequence data for a plasmid derived from this plasmid suggests that the Tet marker was cloned in the reverse orientation to that shown here. Sequencing of the Registry clone would likely confirm this, at which point the sequence of this molecule should be changed.

Source

Austin Che constructed this plamid from pSB1AT3-P1010.