Precise cut-and-paste DNA insertion using engineered type V-K CRISPR-associated transposases.
Tou CJ, Orr B, Kleinstiver BP
Nat Biotechnol. 2023 Jan 2. doi: 10.1038/s41587-022-01574-x.
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Article
Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 181781 | pY2ShHELIX_sgRNA_entry (CJT30) | Expresses Y2-ShHELIX containing a nicking Y2 I-AniI fusion to ShTnsB. New sgRNA spacer sequences can be added using Golden Gate assembly (via SapI sites). |
| 181782 | pShHELIX_sgRNA_entry (CJT32) | Expresses ShHELIX containing a nicking I-AniI fusion to ShTnsB. New sgRNA spacer sequences can be added using Golden Gate assembly (via SapI sites). |
| 181783 | pAcHELIX-sgRNA_entry (CJT94) | Expresses AcHELIX containing a nicking I-AniI fusion to AcTnsB. New sgRNA spacer sequences can be added using Golden Gate assembly (via SapI sites). |
| 181784 | pShoHELIX-sgRNA_entry (BO3) | Expresses ShoHELIX containing a nicking I-AniI fusion to ShoTnsB. New sgRNA spacer sequences can be added using Golden Gate assembly (via SapI sites). |
| 181785 | pAcCAST-sgRNA_entry (CJT83) | Expresses AcCAST. New sgRNA spacer sequences can be added using Golden Gate assembly (via SapI sites). |
| 181786 | pShoCAST-sgRNA_entry (BO1) | Expresses ShoCAST. New sgRNA spacer sequences can be added using Golden Gate assembly (via SapI sites). |
| 181787 | pShCAST(Cas12k-TniQ)-sgRNA_entry (CJT11) | Expresses 3-component ShCAST containing a Cas12k-TniQ fusion. New sgRNA spacer sequences can be added using Golden Gate assembly (via SapI sites). |
| 181788 | pShCAST(Cas12k-TniQ-TniQ)-sgRNA_entry (CJT12) | Expresses 3-component ShCAST containing a Cas12k-TniQ-TniQ fusion. New sgRNA spacer sequences can be added using Golden Gate assembly (via SapI sites). |
| 181789 | pShCAST(Cas12k-TnsC)-sgRNA_entry (CJT28) | Expresses 3-component ShCAST containing a Cas12k-TnsC fusion. New sgRNA spacer sequences can be added using Golden Gate assembly (via SapI sites). |
| 181790 | pShHELIX(Cas12k-TniQ)-sgRNA_entry (CJT111) | Expresses 3-component ShHELIX containing a Cas12k-TniQ fusion. New sgRNA spacer sequences can be added using Golden Gate assembly (via SapI sites). |
| 181791 | pShHELIX(Cas12k-TniQ-TniQ)-sgRNA_entry (CJT112) | Expresses 3-component ShHELIX containing a Cas12k-TniQ-TniQ fusion. New sgRNA spacer sequences can be added using Golden Gate assembly (via SapI sites). |
| 181792 | pShHELIX(Cas12k-TnsC)-sgRNA_entry (CJT113) | Expresses 3-component ShHELIX containing a Cas12k-TnsC fusion. New sgRNA spacer sequences can be added using Golden Gate assembly (via SapI sites). |
| 181793 | pDonor14-ShHELIX-KanR (CJT70) | pDonor for ShHELIX containing 14bp of spacing between the I-AniI site and LE/RE and encoding a kanamycin resistance gene as cargo |
| 181794 | pDonor14-AcHELIX-KanR (CJT96) | pDonor for AcHELIX containing 14bp of spacing between the I-AniI site and LE/RE using ShCAST flanking sequence and encoding a kanamycin resistance gene as cargo |
| 181795 | pDonor14-ShoHELIX-KanR (BO4) | pDonor for ShoHELIX containing 14bp of spacing between the I-AniI site and LE/RE using ShCAST flanking sequence and encoding a kanamycin resistance gene as cargo |
| 190661 | pN7CAST | Expresses polycistronic N7 CAST where Cas12k-T2A-TnsC and TniQ-T2A-TnsB are separated by an EMCV IRES |
| 190662 | pN7HELIX | Expresses polycistronic N7 HELIX where Cas12k-T2A-TnsC and TniQ-T2A-nAniI_TnsB are separated by an EMCV IRES |
| 190663 | pDonor14-N7HELIX-KanR | Kanamycin resistance donor for N7HELIX containing 14bp of spacing between the I-AniI site and LE/RE using ShCAST flanking sequence |
| 190664 | pU6_N7-sgRNA2 | Expresses U6-driven N7 sgRNA2 (poly-T mutated out), BsmbI dropout cassette for cloning |
| 190665 | pN7-S15 | Expresses N7 ribosomal protein S15 |