一 质粒简介：
1. 真核细胞基因敲除/基因编辑
用CRISPR-Cas12f的DNA靶向切割活性，在哺乳动物细胞内完成内源基因敲除、基因片段敲入、位点修饰；
Cas12f体积极小，特别适配腺相关病毒AAV载体的体内递送，适合体内动物基因编辑。
2. 紧凑型CRISPR工具基础研究
质粒可用来研究小型CRISPR系统的编辑效率、靶向特异性、PAM偏好，优化新一代基因编辑工具。
3. 蛋白表达与功能验证
可在细胞内直接表达enAsCas12f融合蛋白，验证蛋白稳定性、定位、酶活，也可用于体外Cas蛋白表达纯化、体外切割实验。
- 序列：
- ATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTAT
- TTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGGCTACCACGCT
- CACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCC
- GGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCA
- GCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCG
- CAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCT
- GAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTT
- CTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCG
- TTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAA
- GCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTG
- ACGTCGACGGATCGGGAGATCGATCTCCCGATCCCCTAGGGTCGACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGTATCTGCTCCCTGCTTGTG
- TGTTGGAGGTCGCTGAGTAGTGCGCGAGCAAAATTTAAGCTACAACAAGGCAAGGCTTGACCGACAATTGCATGAAGAATCTGCTTAGGGTTAGGCGTTTTGCGC
- TGCTTCGCGATGTACGGGCCAGATATACGCGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGT
- TCCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAG
- GGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATG
- ACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGC
- GGTTTTGGCAGTACATCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATC
- AACGGGACTTTCCAAAATGTCGTAACAACTCCGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTGGTTTAGTGAACC
- GTCAGATCCGCTAGAGATCCGCGGCCGCTAATACGACTCACTATAGGGAGAGCCGCCACCATGAAACGGACAGCCGACGGAAGCGAGTTCGAGTCACCAAAGAAG
- AAGCGGAAAGTCATGATCAAGGTGTACAGATACGAGATCGTGAAGCCCCTGGACCTGGATTGGAAGGAATTCGGCACCATCCTGAGACAGCTGCAACAAGAAACC
- AGATTCGCCCTGAACAAGGCCACACAGCTGGCCTGGGAGTGGATGGGCTTCAGCAGCGACTACAAGGACAACCATGGCGAGTACCCTAAGAGCAAGGACATCCTG
- GGCTACACCAATGTGCACGGCTACGCCTACCACACAATCAAAACAAAGGCCTACAGACTGAACAGCGGCAATCTGTCCCAGACCATCAAGCGGGCAACCGACCGG
- TTTAAGGCTTATCAGAAAGAAATCCTGCGGGGCGACATGTCCATCCCCAGCTACAAGCGGGACATTCCTCTGGACCTGATCAAGGAGAACATCTCTGTGAACAGA
- ATGAACCACGGCGATTACATCGCCAGCCTGAGCCTGCTGAGCAACCCCGCCAAGCAGGAGATGAACGTGAAGCGGAAGATCAGCGTGATCATCATAGTTCGGGGT
- GCTGGAAAGACCATCATGGACAGAATCCTCAGCGGCGAATACCAGGTGTCCGCCTCTCAGATCATCCACGACAAGAGAAAGAAGAAATGGTACCTGAATATTTCT
- TATGATTTCGAGCCTCAGACAAGAGTGCTGGATCTCAACAAGATTATGGGCATCGACCTGGGCGTGGCCGTGGCTGTGTACATGGCTTTTCAGCACACCCCTGCC
- AGATACAAGCTGGAAGGCGGAGAGATCGAGAACTTCCGTAGGCAGGTCGAGTCCCGCAGAATCAGCATGCTGCGGCAGGGCAAGTACGCCGGCGGCGCCAGAAGA
- GGCCACGGCAGAGATAAGCGGATCAAGCCTATCGAGCAGCTGAGAGATAAGATCGCTAACTTCAGAGACACCACCAACCACAGATACAGCCGGTACATCGTCGAC
- ATGGCCATCAAGGAGGGCTGCGGCACAATCCAGATGGAAGATCTGACCGGCATCAGAGATATTGGATCTAGATTCCTGCAGAACTGGACCTACTACGACCTGCAG
- CAGAAGATCATCTACAAAGCCGAGGAAGCCGGCATCAAAGTGATCAAAATTAGACCTCAATACACAAGCCAGCGGTGCAGCGAGTGCGGCAACATCGACAGCGGA
- AATAGAATCGGCCAGGCCATCTTCAAGTGCAGAGCGTGTGGATATGAGGCCAATGCCGACTACAACGCCGCCCGCAACATCGCTATCCCAAACATCGACAAGATC
- ATCGCCGAAAGCATCAAAAAAAGAACCGCCGACGGCAGCGAATTCGAGCCCAAGAAGAAGAGGAAAGTCTAACCGGTCATCATCACCATCACCATTGAGTTTAAA
- CCCGCTGATCAGCCTCGACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTT
- CCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATA
- GCAGGCATGCTGGGGATGCGGTGGGCTCTATGGCTTCTGAGGCGGAAAGAACCAGCTGGGGCTCGATACCGTCGACCTCTAGCTAGAGCTTGGCGTAATCATGGT
- CATAGCTGTTTCCTGTGTGAAATTGTTATCCGCTCACAATTCCACACAACATACGAGCCGGAAGCATAAAGTGTAAAGCCTAGGGTGCCTAATGAGTGAGCTAAC
- TCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTA
- TTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAG
- AATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCC
- CTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTC
- CTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGG
- TCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACT
- TATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAA
- GAACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTT
- TTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACACTCAGTGGAACGAAAACTCACGTTAAG
- GGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAA
