LOCUS       sequence-390155-        2310 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pegRNA used to induce GBA (c. 1226 A > G) mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2310)
  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 2310)
  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-GBA-10
FEATURES             Location/Qualifiers
     source          1..2310
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        16..264
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     16..36
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     187..206
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        286..361
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(495..1083)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(575..594)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1254..2045,2046..2114))
                     /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     1877..1896
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2115..2219)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaggtcgggc aggaagaggg cctatttccc atgattcctt catatttgca tatacgatac
       61 aaggctgtta gagagataat tagaattaat ttgactgtaa acacaaagat attagtacaa
      121 aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt
      181 aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat
      241 atcttgtgga aaggacgaaa caccgagccg accacatggt acagggtttt agagctagaa
      301 atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtgggac cgagtcggtc
      361 cttaccctag agcctcctgt accatgtggt ctttttttaa gcttgggccg ctcgaggtac
      421 ctctctacat atgacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
      481 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
      541 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
      601 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
      661 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
      721 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
      781 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
      841 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
      901 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
      961 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     1021 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     1081 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     1141 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     1201 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     1261 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     1321 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     1381 gcaatgatac cgcgagatcc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     1441 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     1501 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     1561 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     1621 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     1681 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     1741 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     1801 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     1861 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     1921 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     1981 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     2041 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     2101 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     2161 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     2221 acatttcccc gaaaagtgcc acctgacgtc gctagctgta caaaaaagca ggctttaaag
     2281 gaaccaattc agtcgactgg atccggtacc
//
