Difference between revisions of "Part:BBa K1807000:Design"
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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.
 
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.
  
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The Overhang Sequences are as follows:  
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The overhang sequences used to make this part are as follows:  
  
 
'''BBa_K1807000 Assembly''' 5'end Overhang (complementary to pSB1C3): cgctaaggatgatttctgGAATTCGCGGCCGCTTCTAGAG
 
'''BBa_K1807000 Assembly''' 5'end Overhang (complementary to pSB1C3): cgctaaggatgatttctgGAATTCGCGGCCGCTTCTAGAG
  
 
'''BBa_K1807000 Assembly''' 3'end Overhang (complementary to pSB1C3): TACTAGTAGCGGCCGCTGCAGtccggcaaaaaagggcaag
 
'''BBa_K1807000 Assembly''' 3'end Overhang (complementary to pSB1C3): TACTAGTAGCGGCCGCTGCAGtccggcaaaaaagggcaag
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The '''Splice-In Flanking Sequences''' that allow any Protein Coding Sequence to be inserted in place of lacZ alpha:
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5' end: '''v'''
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3' end: '''v'''
  
  

Revision as of 16:11, 18 September 2015

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 used to make this part are as follows:

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

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

The Splice-In Flanking Sequences that allow any Protein Coding Sequence to be inserted in place of lacZ alpha:

5' end: v

3' end: v


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