LOCUS       sequence-390157-        2305 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pegRNA used to induce LMNA (c.1824C > T) mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2305)
  AUTHORS   Li M, Zhong A, Wu Y, Sidharta M, Beaury M, Zhao X, Studer L, Zhou T
  TITLE     Transient inhibition of p53 enhances prime editing and cytosine 
            base-editing efficiencies in human pluripotent stem cells.
  JOURNAL   Nat Commun. 2022 Oct 27;13(1):6354. doi: 10.1038/s41467-022-34045-7.
  PUBMED    36302757
REFERENCE   2  (bases 1 to 2305)
  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: "Nat 
            Commun."; date: "2022-10-27"; volume: "13(1):6354. doi"; pages: " 
            10.1038/s41467-022-34045-7"
COMMENT     Sequence Label: pegRNA_LMNA-1
FEATURES             Location/Qualifiers
     source          1..2305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..275
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     27..47
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     198..217
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        297..372
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(501..1089)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(581..600)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1260..2051,2052..2120))
                     /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     1883..1902
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2121..2225)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gatccggtac caaggtcggg caggaagagg gcctatttcc catgattcct tcatatttgc
       61 atatacgata caaggctgtt agagagataa ttagaattaa tttgactgta aacacaaaga
      121 tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa
      181 aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct
      241 tggctttata tatcttgtgg aaaggacgaa acaccgggct caggagccca ggtggggttt
      301 tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggga
      361 ccgagtcggt ccatgggtcc acccacctgg gctcctgttt ttttaagctt gggccgctcg
      421 aggtacctct ctacatatga catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
      481 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
      541 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
      601 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
      661 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
      721 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
      781 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
      841 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
      901 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
      961 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     1021 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     1081 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     1141 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     1201 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     1261 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     1321 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     1381 agtgctgcaa tgataccgcg agatccacgc tcaccggctc cagatttatc agcaataaac
     1441 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     1501 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     1561 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     1621 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     1681 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     1741 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     1801 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     1861 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     1921 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     1981 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     2041 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     2101 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     2161 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     2221 ccgcgcacat ttccccgaaa agtgccacct gacgtcgcta gctgtacaaa aaagcaggct
     2281 ttaaaggaac caattcagtc gactg
//
