Part:BBa_K2201280

spacer with BseRI recognition site

Recursive Directional Ligation by Plasmid (PRe-RDL) uses three restriction sites of a parent plasmid which contains the gene of interest (goi) and the subsequently ligation of fragments of two different restricted parent plasmids. The first step involves the restriction of a parent plasmid at the 3'-end of the goi and in the backbone. The second step is the restriction of a parent plasmid at the 5'-end of the goi and at the same position of the backbone as in the first step. The final step is the ligation of both generated fragments containing the goi. The result is a plasmid with two copies of the goi. Applied to pSB1C3 containing the consensus sequences of monomers (like K2201282 or K2201283) and the spacers K2201280 and K2201281 between the BioBrick prefix and suffix shown in following figure, it is possible to use the PRe-RDL in combination with the restriction enzymes, AcuI, BseRI, and BspEI, to build repetitive sequences of monomers.

Design of K2201250 and K2201251.

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
  COMPATIBLE WITH RFC[1000]