Part:BBa_K1807000:Design

Above is a diagram of our Adapter BioBrick in the pSB1C3 vector. Our design incorporates a SmaI restriction digestion step and subsequent use of the cut Adapter as a vector in a Gibson Assembly Reaction or any other assembly method based on flanking overlapping sequences (as depicted below).

BBa_K1807000 was designed as a blue-white screening device that would also be easily used in Gibson Assembly. Surrounding the lacZ alpha coding sequence are two SmaI restriction sites (CCC/GGG). SmaI is a blunt-end endonuclease- we used it to simulatenously linearize our vector and remove the lacZ alpha coding sequence. Our BioBricks contained overhangs that make them compatible with the SmaI-digested BBa_K1807000.

The Overhang Sequences are as follows:

BBa_K1807000 Assembly 5'end Overhang (complementary to pSB1C3): cgctaaggatgatttctgGAATTCGCGGCCGCTTCTAGAG

BBa_K1807000 Assembly 3'end Overhang (complementary to pSB1C3): TACTAGTAGCGGCCGCTGCAGtccggcaaaaaagggcaag

The subparts of the device are as follows: BBa_R0011, BBa_B0034, BBa_E0038, BBa_B0015.