Part:BBa_B0103

SapI restriction enzyme site, reversed (offset cutter)

SapI restriction site in the reverse orientation (recognition sequence on the 3' strand). SapI is an offsite cutter. From left to right this part includes the designed 3 bp 5' overhang (ATG), a single bp spacer, the 7 bp recognition sequence (5'-GCTCTTCT-3'), and two bp spacer. Cleavage at this site results in a 3 bp, 5' overhang (left side, top strand), which makes it compatible with Heather Keller's RBS assembly scheme for creating RBS/CDS junctions without a mixed site. This part is specific for CDSs with an ATG start site and is not compatible with GTG start sites (via HKs assembly scheme). The single bp spacer on the 3' end allows RBSes construct with this part to be used in standard biobricks assembly and is necessary to maintain the reading frame of a downstream CDS. Note that Standard Biobricks Assembly with this site results in the creation of a new ATG start site, and a 7 amino acid fusion at the N terminus of the downstream protein.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 5