Part:BBa_K2649002:Design

RFP SB1C3 w/ SapI

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 247
  Illegal XbaI site found at 220
  Illegal PstI site found at 208
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 247
  Illegal PstI site found at 208
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 247
  Illegal BamHI site found at 226
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 247
  Illegal XbaI site found at 220
  Illegal PstI site found at 208
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 247
  Illegal XbaI site found at 220
  Illegal PstI site found at 208
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal BsaI site found at 1
  Illegal BsaI.rc site found at 598
  

Design Notes

Primer Information

Forward (iGEM 46 RFP SapI Primer): gaattcgcggccgcttctagagagcggaagagccaatacgcaaaccgcc

Reverse (iGEM 40 RFP SapI Reverse Primer): tactagtcactgaagagctataaacgcagaaaggccc

Tm: 62 degrees for both primers

PCR at 63 degrees - Adds the SapI binding and cut sites to the beginning and end of the RFP construct to allow for insertion of Lab parts and adds certain BioBrick sites (EcoRI, NotI, and XbaI to the beginning, SpeI to the end) for insertion into the iGEM SB1C3 backbone. XbaI and SpeI have the same sticky ends, so had to extend to use whole prefix.