Part:BBa_K3001023
Lsh crRNA targeting RNA Mango with T7 promoter
This CRISPR RNA (crRNA) is designed to interact with Cas13a isolated from Leptotrichia shahii (Lsh). It is designed to target snR30-RNA Mango II RNA transcript. It is designed to be in vitro transcribed using T7 polymerase. This construct was synthesized in pUC19.
Wetlab data
CRISPR RNA (crRNA) is a crucial aspect of the CRISPR Cas13a system. It seeks out the target RNA sequence so that the Cas13a can cleave the RNA. Our crRNA sequences were synthesized by IDT into the plasmid pUC19. Then we successfully PCR amplified the DNA out of the pUC19 plasmid.
Figure 1. 10% DNA PAGE of crRNA products for Lsh and Lwa. Left to right: lane 1: 100 bp ladder (Thermo Scientific. Lane 2: Lwa 56.5; Lane 3: Lwa 56.8; lane 4: Lsh 56.5; lane 5: Lsh 56.8. Note that all lanes are from the same gel.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 67
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 67
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 67
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 67
- 1000COMPATIBLE WITH RFC[1000]
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