LOCUS       sequence-354425-        2302 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  S. pyogenes pegRNA for +5 G to T edit at the RNF2 site of human 
            cells using prime editing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2302)
  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 2302)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            2019 Oct 21. pii: 10.1038/s41586-019-1711-4. doi: 
            10.1038/s41586-019-1711-4."
COMMENT     Sequence Label: pU6-Sp-pegRNA-RNF2_+5GtoT
FEATURES             Location/Qualifiers
     source          1..2302
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(31..822,823..891))
                     /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     654..673
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(892..996)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1103..1351
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1103..1123
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1274..1293
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        1372..1447
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(1574..2162)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1654..1673)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
       61 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
      121 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagatccacg
      181 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
      241 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
      301 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
      361 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
      421 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
      481 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
      541 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
      601 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
      661 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
      721 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
      781 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
      841 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
      901 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
      961 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     1021 tgacgtcgct agctgtacaa aaaagcaggc tttaaaggaa ccaattcagt cgactggatc
     1081 cggtaccaag gtcgggcagg aagagggcct atttcccatg attccttcat atttgcatat
     1141 acgatacaag gctgttagag agataattag aattaatttg actgtaaaca caaagatatt
     1201 agtacaaaat acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt
     1261 atgttttaaa atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc
     1321 tttatatatc ttgtggaaag gacgaaacac cgtcatctta gtcattacct ggttttagag
     1381 ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag
     1441 tcggtgcaac gaacacatca ggtaatgact aagatgtttt tttgggccgc tcgaggtacc
     1501 tctctacata tgacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1561 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1621 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     1681 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     1741 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     1801 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     1861 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     1921 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     1981 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     2041 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2101 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2161 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2221 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     2281 aatgaagttt taaatcaatc ta
//
