Part:BBa_K2961006

Template DNA sequence coding for HBB rs334 mutation (A to T) in humans

This sequence encodes for the HBB gene with the rs334 mutation, which is the most common mutation that leads to the sickle cell anemia phenotype (Glu6Val). One of the nucleotides of the gene was modified so that the Cpf1 PAM site could be added adjacent to a target site for our gRNA, but the overall template largely remains the same.

This part is a direct improvement on a part designed last year with the same intention (BBa_K2872881). This year's edition only modifies one nucleotide instead of adding four nucleotides artificially like the previous one. Furthermore, the PAM site is directly adjacent to the target gRNA sequence that was designed for this template, which is more appropriate for Cpf1, compared to last year where the PAM site was 20 nucleotides away from the unique target site that distinguished the wildtype and mutant alleles.

We have characterized this part but have failed to see any downstream fluorescence effects that we were hoping to observe.

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 BsaI.rc site found at 174