LOCUS       sequence-390156-        2307 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pegRNA used to induce LRRK2 (c. 6055 G > A) mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2307)
  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 2307)
  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_LRRK2-3
FEATURES             Location/Qualifiers
     source          1..2307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     414..433
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(652..756)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        863..1111
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     863..883
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1034..1053
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        1133..1208
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(1339..1927)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1419..1438)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2098..2307,1..582,583..651))
                     /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"
ORIGIN
        1 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
       61 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
      121 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
      181 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
      241 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
      301 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
      361 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
      421 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
      481 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
      541 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
      601 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
      661 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
      721 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
      781 tgacgtcgct agctgtacaa aaaagcaggc tttaaaggaa ccaattcagt cgactggatc
      841 cggtaccaag gtcgggcagg aagagggcct atttcccatg attccttcat atttgcatat
      901 acgatacaag gctgttagag agataattag aattaatttg actgtaaaca caaagatatt
      961 agtacaaaat acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt
     1021 atgttttaaa atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc
     1081 tttatatatc ttgtggaaag gacgaaacac cgattgcaaa gattgctgac tagttttaga
     1141 gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtgggaccga
     1201 gtcggtccag caatgctgta gtcagcaatc tttgcttttt ttaagcttgg gccgctcgag
     1261 gtacctctct acatatgaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1321 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1381 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1441 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1501 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1561 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1621 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1681 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1741 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     1801 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1861 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1921 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1981 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2041 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2101 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2161 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     2221 tgctgcaatg ataccgcgag atccacgctc accggctcca gatttatcag caataaacca
     2281 gccagccgga agggccgagc gcagaag
//
