LOCUS       sequence-416936-        2380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dnmt1 targeting epegRNA with MS2 insertion in gRNA scaffold for v3 
            PE-eVLP production.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2380)
  AUTHORS   An M, Raguram A, Du SW, Banskota S, Davis JR, Newby GA, Chen PZ, 
            Palczewski K, Liu DR
  TITLE     Engineered virus-like particles for transient delivery of prime 
            editor ribonucleoprotein complexes in vivo.
  JOURNAL   Nat Biotechnol. 2024 Jan 8. doi: 10.1038/s41587-023-02078-y.
  PUBMED    38191664
REFERENCE   2  (bases 1 to 2380)
  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 
            Biotechnol. 2024 Jan 8. doi: 10.1038/s41587-023-02078-y."
COMMENT     Sequence Label: MS2-epegRNA-Dnmt1
FEATURES             Location/Qualifiers
     source          1..2380
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        82..330
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     82..102
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     253..272
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        418..436
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     rep_origin      complement(631..1219)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(711..730)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1390..2181,2182..2250))
                     /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     2013..2032
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2251..2355)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcgcta gctgtacaaa aaagcaggct ttaaaggaac caattcagtc gactggatcc
       61 ggtaccaagg tcgggcagga agagggccta tttcccatga ttccttcata tttgcatata
      121 cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta
      181 gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta
      241 tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct
      301 ttatatatct tgtggaaagg acgaaacacc cgggctggag ctgttcgcgc gtttaagagc
      361 tatgctggaa acagcatagc aagtttaaat aaggctagtc cgttatcaac ttggccaaca
      421 tgaggatcac ccatgtctgc agggccaagt ggcaccgagt cggtgctgca agatgcgttg
      481 gcgaacagct ccaggataat atcgcggttc tatctagtta cgcgttaaac caactagaat
      541 tttttaagct tgggccgctc gaggtacctc tctacatatg acatgtgagc aaaaggccag
      601 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
      661 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
      721 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
      781 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
      841 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
      901 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
      961 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     1021 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     1081 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     1141 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     1201 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     1261 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     1321 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     1381 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     1441 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     1501 atctggcccc agtgctgcaa tgataccgcg agatccacgc tcaccggctc cagatttatc
     1561 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     1621 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     1681 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     1741 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     1801 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     1861 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     1921 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     1981 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     2041 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     2101 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     2161 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     2221 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     2281 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     2341 aataggggtt ccgcgcacat ttccccgaaa agtgccacct
//
