Part:BBa_K4058025:Design
ICE2 sgRNA C in pHSN6A01
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 7833
Illegal XbaI site found at 2961
Illegal SpeI site found at 9996
Illegal PstI site found at 2032
Illegal PstI site found at 3326
Illegal PstI site found at 3872
Illegal PstI site found at 5294
Illegal PstI site found at 5498
Illegal PstI site found at 6740
Illegal PstI site found at 12904 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 7833
Illegal NheI site found at 4538
Illegal SpeI site found at 9996
Illegal PstI site found at 2032
Illegal PstI site found at 3326
Illegal PstI site found at 3872
Illegal PstI site found at 5294
Illegal PstI site found at 5498
Illegal PstI site found at 6740
Illegal PstI site found at 12904
Illegal NotI site found at 2953 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 7833
Illegal BglII site found at 4128
Illegal BglII site found at 4818
Illegal BamHI site found at 2930
Illegal BamHI site found at 4422
Illegal BamHI site found at 6460
Illegal BamHI site found at 7557
Illegal XhoI site found at 2922
Illegal XhoI site found at 3746
Illegal XhoI site found at 4928
Illegal XhoI site found at 6092
Illegal XhoI site found at 8657
Illegal XhoI site found at 9751 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 7833
Illegal XbaI site found at 2961
Illegal SpeI site found at 9996
Illegal PstI site found at 2032
Illegal PstI site found at 3326
Illegal PstI site found at 3872
Illegal PstI site found at 5294
Illegal PstI site found at 5498
Illegal PstI site found at 6740
Illegal PstI site found at 12904 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 7833
Illegal XbaI site found at 2961
Illegal SpeI site found at 9996
Illegal PstI site found at 2032
Illegal PstI site found at 3326
Illegal PstI site found at 3872
Illegal PstI site found at 5294
Illegal PstI site found at 5498
Illegal PstI site found at 6740
Illegal PstI site found at 12904
Illegal NgoMIV site found at 1457
Illegal NgoMIV site found at 4690
Illegal NgoMIV site found at 10733
Illegal AgeI site found at 9030 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1746
Illegal BsaI.rc site found at 533
Illegal SapI site found at 3664
Illegal SapI.rc site found at 5131
Design Notes
pHSN6A01 was a gift from Qi-Jun Chen (Addgene plasmid # 50586 ; http://n2t.net/addgene:50586 ; RRID:Addgene_50586)
The gRNA sequence was designed using the guide RNA design software CHOPCHOP. The parameters were set to find optimal CRISPR/Cas9 target sites for CRISPR activation. Target sequences were selected based on the predicted binding efficiency provided by CHOPCHOP.
Source
pHSN6A01 was a gift from Qi-Jun Chen (Addgene plasmid # 50586 ; http://n2t.net/addgene:50586 ; RRID:Addgene_50586)
References
A CRISPR/Cas9 toolkit for multiplex genome editing in plants. Xing HL, Dong L, Wang ZP, Zhang HY, Han CY, Liu B, Wang XC, Chen QJ. BMC Plant Biol. 2014 Nov 29;14(1):327. 10.1186/s12870-014-0327-y PubMed 25432517
Labun, K., Montague, T. G., Krause, M., Torres Cleuren, Y. N., Tjeldnes, H., & Valen, E. CHOPCHOP v3: expanding the CRISPR web toolbox beyond genome editing. Nucleic Acids Research (2019).