Other

Part:BBa_K3208012

Designed by: Slavil Peykov   Group: iGEM19_Bulgaria   (2019-10-21)
Revision as of 00:11, 22 October 2019 by Registry (Talk | contribs)

(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)


Modified gRNA expression vector

This part is a modified version of part BBa_K3208000. This new gRNA expression vector contains red expression cassette (BBa_ K3208010) inserted between the two Eco31I sites. This allows selection of positive transformants based on colour when one uses these 2 sites for cloning of new gRNAs.

We performed 3 cloning experiments (with 3 different gRNAs that were given to us as overlapping oligos by team iGEM Bulgaria 2017) ) and we found that when using the right color as a selection tool, we increased the cloning efficiency to the range of 4-6 positive colonies per 10 white colonies. This is a huge improvement compared to the previous version (see part BBa_K2515002 for details).


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 8
    Illegal NheI site found at 31
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 824
    Illegal AgeI site found at 936
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1073
    Illegal BsaI.rc site found at 38


[edit]
Categories
Parameters
None