LOCUS       sequence-390158-        2310 bp    DNA     circular SYN 14-APR-2025
DEFINITION  pegRNA used to correct 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 Apr 14, 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-4
FEATURES             Location/Qualifiers
     source          1..2310
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        12..260
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     12..32
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     183..202
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        282..357
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(491..1079)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(571..590)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1250..2041,2042..2110))
                     /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     1873..1892
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2111..2215)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgggcagga agagggccta tttcccatga ttccttcata tttgcatata cgatacaagg
       61 ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta gtacaaaata
      121 cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa
      181 tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct
      241 tgtggaaagg acgaaacacc gagccgacca catggtacag ggttttagag ctagaaatag
      301 caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tgggaccgag tcggtcctta
      361 ccctagaacc tcctgtacca tgtggtcttt ttttaagctt gggccgctcg aggtacctct
      421 ctacatatga catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
      481 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
      541 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
      601 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
      661 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
      721 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
      781 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
      841 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
      901 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
      961 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     1021 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     1081 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     1141 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     1201 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     1261 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     1321 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     1381 tgataccgcg agatccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     1441 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     1501 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     1561 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     1621 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     1681 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     1741 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     1801 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     1861 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     1921 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     1981 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     2041 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     2101 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     2161 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     2221 ttccccgaaa agtgccacct gacgtcgcta gctgtacaaa aaagcaggct ttaaaggaac
     2281 caattcagtc gactggatcc ggtaccaagg
//
