Difference between revisions of "Part:BBa K3982001"
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To know more about how this part is used in project CODE M, visit our wikipage. [https://2021.igem.org/Team:IISER_Berhampur/Team IISER_Berhampur 2021] | To know more about how this part is used in project CODE M, visit our wikipage. [https://2021.igem.org/Team:IISER_Berhampur/Team IISER_Berhampur 2021] | ||
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+ | ===References=== | ||
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+ | 1) Harrington, L. B., Burstein, D., Chen, J. S., Paez-Espino, D., Ma, E., Witte, I. P., Cofsky, J. C., Kyrpides, N. C., Banfield, J. F., & Doudna, J. A. (2018). Programmed DNA destruction by miniature CRISPR-Cas14 enzymes. Science (New York, N.Y.), 362(6416), 839–842. https://doi.org/10.1126/science.aav4294 | ||
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+ | 2) Takeda, S. N., Nakagawa, R., Okazaki, S., Hirano, H., Kobayashi, K., Kusakizako, T., Nishizawa, T., Yamashita, K., Nishimasu, H., & Nureki, O. (2021). Structure of the miniature type V-F CRISPR-Cas effector enzyme. Molecular cell, 81(3), 558–570.e3. https://doi.org/10.1016/j.molcel.2020.11.035 | ||
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+ | 3) Karvelis, T., Bigelyte, G., Young, J. K., Hou, Z., Zedaveinyte, R., Budre, K., Paulraj, S., Djukanovic, V., Gasior, S., Silanskas, A., Venclovas, Č., & Siksnys, V. (2020). PAM recognition by miniature CRISPR-Cas12f nucleases triggers programmable double-stranded DNA target cleavage. Nucleic acids research, 48(9), 5016–5023. https://doi.org/10.1093/nar/gkaa208 | ||
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Revision as of 16:03, 12 October 2021
Cas14a1 expression gene
This sequence encodes the Cas14a1 protein. It is a highly compact RNA guided nuclease used by Team IISER_Berhampur 2021 in Project CODE M.
Usage and Biology
Cas14a1 (also called Un1Cas12f1) is a miniature cas protein used in CRISPR/Cas technology identified from archaebacteria and bacteriophages. It is a RNA guided nuclease that cleaves ssDNA without any Protospacer Adjacent Motif (PAM) specificity i.e. PAM independent ssDNA cleavage. This enables high-fidelity Single-nucleotide polymorphism (SNP) genotyping. It is guided by a single guide RNA (sgRNA) which has two components - CRISPR RNA (crRNA) and Trans-activating crispr RNA (tracrRNA).
To know more about how this part is used in project CODE M, visit our wikipage. IISER_Berhampur 2021
References
1) Harrington, L. B., Burstein, D., Chen, J. S., Paez-Espino, D., Ma, E., Witte, I. P., Cofsky, J. C., Kyrpides, N. C., Banfield, J. F., & Doudna, J. A. (2018). Programmed DNA destruction by miniature CRISPR-Cas14 enzymes. Science (New York, N.Y.), 362(6416), 839–842. https://doi.org/10.1126/science.aav4294
2) Takeda, S. N., Nakagawa, R., Okazaki, S., Hirano, H., Kobayashi, K., Kusakizako, T., Nishizawa, T., Yamashita, K., Nishimasu, H., & Nureki, O. (2021). Structure of the miniature type V-F CRISPR-Cas effector enzyme. Molecular cell, 81(3), 558–570.e3. https://doi.org/10.1016/j.molcel.2020.11.035
3) Karvelis, T., Bigelyte, G., Young, J. K., Hou, Z., Zedaveinyte, R., Budre, K., Paulraj, S., Djukanovic, V., Gasior, S., Silanskas, A., Venclovas, Č., & Siksnys, V. (2020). PAM recognition by miniature CRISPR-Cas12f nucleases triggers programmable double-stranded DNA target cleavage. Nucleic acids research, 48(9), 5016–5023. https://doi.org/10.1093/nar/gkaa208
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 1357
Illegal PstI site found at 147
Illegal PstI site found at 313 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1357
Illegal PstI site found at 147
Illegal PstI site found at 313 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1357
Illegal BamHI site found at 951 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 1357
Illegal PstI site found at 147
Illegal PstI site found at 313 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1357
Illegal PstI site found at 147
Illegal PstI site found at 313
Illegal AgeI site found at 607 - 1000COMPATIBLE WITH RFC[1000]