LOCUS       sequence-390159-        2328 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pegRNA used to correct LMNA (c.1824C > T) mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2328)
  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 2328)
  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_CR-2-5
FEATURES             Location/Qualifiers
     source          1..2328
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        20..268
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     20..40
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     191..210
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        290..365
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(517..1105)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(597..616)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1276..2067,2068..2136))
                     /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     1899..1918
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2137..2241)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 taccaaggtc gggcaggaag agggcctatt tcccatgatt ccttcatatt tgcatatacg
       61 atacaaggct gttagagaga taattagaat taatttgact gtaaacacaa agatattagt
      121 acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg
      181 ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt
      241 atatatcttg tggaaaggac gaaacaccgg ccgacaaggc atctgccagg ttttagagct
      301 agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg ggaccgagtc
      361 ggtccagatg ggtccgccca cctgggctcc tgagccgctg gcagatgcct tgtttttttt
      421 aagcttgggc cgctcgaggt acctctctac atatgacatg tgagcaaaag gccagcaaaa
      481 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
      541 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
      601 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
      661 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
      721 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
      781 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
      841 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
      901 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
      961 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1021 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1081 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1141 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     1201 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1261 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     1321 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     1381 cttaccatct ggccccagtg ctgcaatgat accgcgagat ccacgctcac cggctccaga
     1441 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     1501 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     1561 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     1621 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     1681 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     1741 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     1801 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     1861 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     1921 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     1981 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     2041 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     2101 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     2161 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     2221 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgctagctg
     2281 tacaaaaaag caggctttaa aggaaccaat tcagtcgact ggatccgg
//
