Difference between revisions of "Part:BBa K3791014:Design"
Line 7: Line 7:
  
 
===Design Notes===
 
===Design Notes===
−
The repeat sequence must be placed before the spacer sequence, that is, in the 5’ extreme.
+
The repeat sequence must be placed before the spacer sequence, that is, in the 5’ extreme. It is also required to have a length between 18 and 24 nucleotides, in this case adjusting it to a consensus G-C content of approximately 50%.
  
 
===Source===
 
===Source===
  
−
The repeat and the T7 promoter sequences were extracted from the parts registry ([https://parts.igem.org/wiki/index.php?title=Part:BBa_K2927006 BBa_K2927006]) ([https://parts.igem.org/Part:BBa_K1614000 BBa_K1614000]), whereas the spacer one was obtained from being the complementary sequence of an spectinomycin-resistant gene DNA fragment. (as explained in its own part page: [https://parts.igem.org/Part:BBa_K3791000 BBa_K3791004]).
+
The repeat and the T7 promoter sequences were extracted from the parts registry ([https://parts.igem.org/wiki/index.php?title=Part:BBa_K2927006 BBa_K2927006]) ([https://parts.igem.org/Part:BBa_K1614000 BBa_K1614000]), whereas the spacer one was designed with the constraint that it had to be complementary to the sequence of the spectinomycin-resistance gene aiming to be detected (as explained in its own part page: [https://parts.igem.org/Part:BBa_K3791000 BBa_K3791004]).
  
 
===References===
 
===References===
 
[1] New England Biolabs. (s/f). <b>How do I design a guide RNA for use with EnGen Lba Cas12a?</b> Neb.com. From: https://international.neb.com/faqs/2018/05/03/how-do-i-design-a-guide-rna-for-use-with-engen-lba-cas12a
 
[1] New England Biolabs. (s/f). <b>How do I design a guide RNA for use with EnGen Lba Cas12a?</b> Neb.com. From: https://international.neb.com/faqs/2018/05/03/how-do-i-design-a-guide-rna-for-use-with-engen-lba-cas12a

Revision as of 01:21, 13 October 2021


gRNA Spectinomycin construct


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]


Design Notes

The repeat sequence must be placed before the spacer sequence, that is, in the 5’ extreme. It is also required to have a length between 18 and 24 nucleotides, in this case adjusting it to a consensus G-C content of approximately 50%.

Source

The repeat and the T7 promoter sequences were extracted from the parts registry (BBa_K2927006) (BBa_K1614000), whereas the spacer one was designed with the constraint that it had to be complementary to the sequence of the spectinomycin-resistance gene aiming to be detected (as explained in its own part page: BBa_K3791004).

References

[1] New England Biolabs. (s/f). How do I design a guide RNA for use with EnGen Lba Cas12a? Neb.com. From: https://international.neb.com/faqs/2018/05/03/how-do-i-design-a-guide-rna-for-use-with-engen-lba-cas12a