LOCUS Exported 2305 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION S. pyogenes pegRNA for CTT insertion at the HEK3 site of human cells using prime editing. ACCESSION . VERSION . KEYWORDS pU6-Sp-pegRNA-HEK3_CTT_ins SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2305) AUTHORS Anzalone AV, Randolph PB, Davis JR, Sousa AA, Koblan LW, Levy JM, Chen PJ, Wilson C, Newby GA, Raguram A, Liu DR TITLE Search-and-replace genome editing without double-strand breaks or donor DNA. JOURNAL Nature. 2019 Oct 21. pii: 10.1038/s41586-019-1711-4. doi: 10.1038/s41586-019-1711-4. PUBMED 31634902 REFERENCE 2 (bases 1 to 2305) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2305 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 82..322 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 82..102 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 253..272 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_RNA 351..426 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" rep_origin complement(556..1144) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(636..655) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1315..2175) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 1938..1957 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2176..2280) /gene="bla" /label=AmpR promoter ORIGIN 1 gacgtcgcta gctgtacaaa aaagcaggct ttaaaggaac caattcagtc gactggatcc 61 ggtaccaagg tcgggcagga agagggccta tttcccatga ttccttcata tttgcatata 121 cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta 181 gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta 241 tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct 301 ttatatatct tgtggaaagg acgaaacacc ggcccagact gagcacgtga gttttagagc 361 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt gggaccgagt 421 cggtcctctg ccatcaaagc gtgctcagtc tgttttttta agcttgggcc gctcgaggta 481 cctctctaca tatgacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc 541 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg 601 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg 661 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt 721 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt 781 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg 841 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact 901 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt 961 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct 1021 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac 1081 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc 1141 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg 1201 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta 1261 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca 1321 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc 1381 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc 1441 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc 1501 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat 1561 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt 1621 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc 1681 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag 1741 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt 1801 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac 1861 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg 1921 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat 1981 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc 2041 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc 2101 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa 2161 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg 2221 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg 2281 cacatttccc cgaaaagtgc cacct //