LOCUS       sequence-369051-        6998 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  p3-PE2-C-term(1153-dRnH) in AAV2 backbone (ITRs) targeting DNMT1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6998)
  AUTHORS   Bock D, Rothgangl T, Villiger L, Schmidheini L, Matsushita M, Mathis
            N, Ioannidi E, Rimann N, Grisch-Chan HM, Kreutzer S, Kontarakis Z, 
            Kopf M, Thony B, Schwank G
  TITLE     In vivo prime editing of a metabolic liver disease in mice.
  JOURNAL   Sci Transl Med. 2022 Mar 16;14(636):eabl9238. doi: 
            10.1126/scitranslmed.abl9238. Epub 2022 Mar 16.
  PUBMED    35294257
REFERENCE   2  (bases 1 to 6998)
  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: "Sci Transl 
            Med."; date: "2022-03-16"; volume: "14(636):eabl9238. doi"; pages: "
            10.1126/scitranslmed.abl9238. Epub 2022 Mar 16"
COMMENT     Sequence Label: AAV-p3-PE(1153-dRnH)-C-term
FEATURES             Location/Qualifiers
     source          1..6998
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      392..980
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     881..900
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    1268..1289
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1304..1309,1310..1327,1328..1334)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1342..1358
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     1347..1369
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1366..1382
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1366..1382
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   1409..1538
                     /label=AAV2 ITR
     CDS             1918..1938
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4318..4338
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4404..4424)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4615..4736
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4626..4645)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4680..4699
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4743..4991
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     4743..4763
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4914..4933
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        5013..5088
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        5128..5342
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     5263..5282
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     misc_RNA        5372..5447
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     repeat_region   5467..5596
                     /label=AAV2 ITR
     primer_bind     complement(5615..5632)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5615..5631)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5624..5646)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5773..6228
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5860..5879)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6070..6091
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6254..6358
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6359..6427,6428..6998,1..221)
                     /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     complement(6577..6596)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
       61 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      121 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
      181 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
      241 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
      301 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
      361 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
      421 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
      481 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
      541 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
      601 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
      661 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
      721 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
      781 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
      841 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
      901 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
      961 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1021 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1081 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1141 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     1201 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     1261 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     1321 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     1381 accatgatta cgccagattt aattaaggct gcgcgctcgc tcgctcactg aggccgcccg
     1441 ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg
     1501 cagagaggga gtggccaact ccatcactag gggttcctgc ggcccctaga tcagaccgcg
     1561 cgccggggga ggctgctggt gaatattaac caaggtcacc ccagttatcg gaggagcaaa
     1621 caggggctaa gtccacacgc gtggtaccgt ctgtctgcac atttcgtaga gcgagtgttc
     1681 cgatactcta atctccctag gcaaggttca tatttgtgta ggttacttat tctccttttg
     1741 ttgactaagt caataatcag aatcagcagg tttggagtca gcttggcagg gatcagcagc
     1801 ctgggttgga aggagggggt ataaaagccc cttcaccagg agaagccgtc acacagatcc
     1861 acaagctcct gggccgccac catgaaacgg acagccgatg gatctgagtt tgagtctccg
     1921 aagaaaaaac gcaaggtcat caagattgct acacggaaat acctgggaaa gcagaacgtg
     1981 tacgacatcg gcgtggagcg ggatcacaac ttcgcgctga aaaatggatt catagcgagc
     2041 aattccggcg gtggaggctc cggaggcggt ggcagcggag gtggaggtag ctccaagaaa
     2101 ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag
     2161 aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc
     2221 aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct
     2281 gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg
     2341 tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag
     2401 ctgtttgtgg aacagcacaa gcactacctg gacgagatca tcgagcagat cagcgagttc
     2461 tccaagagag tgatcctggc cgacgctaat ctggacaaag tgctgtccgc ctacaacaag
     2521 caccgggata agcccatcag agagcaggcc gagaatatca tccacctgtt taccctgacc
     2581 aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagaggtac
     2641 accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac
     2701 gagacacgga tcgacctgtc tcagctggga ggtgactctg gaggatctag cggaggatcc
     2761 tctggcagcg agacaccagg aacaagcgag tcagcaacac cagagagcag tggcggcagc
     2821 agcggcggca gcagcaccct aaatatagaa gatgagtatc ggctacatga gacctcaaaa
     2881 gagccagatg tttctctagg gtccacatgg ctgtctgatt ttcctcaggc ctgggcggaa
     2941 accgggggca tgggactggc agttcgccaa gctcctctga tcatacctct gaaagcaacc
     3001 tctacccccg tgtccataaa acaatacccc atgtcacaag aagccagact ggggatcaag
     3061 ccccacatac agagactgtt ggaccaggga atactggtac cctgccagtc cccctggaac
     3121 acgcccctgc tacccgttaa gaaaccaggg actaatgatt ataggcctgt ccaggatctg
     3181 agagaagtca acaagcgggt ggaagacatc caccccaccg tgcccaaccc ttacaacctc
     3241 ttgagcgggc tcccaccgtc ccaccagtgg tacactgtgc ttgatttaaa ggatgccttt
     3301 ttctgcctga gactccaccc caccagtcag cctctcttcg cctttgagtg gagagatcca
     3361 gagatgggaa tctcaggaca attgacctgg accagactcc cacagggttt caaaaacagt
     3421 cccaccctgt ttaatgaggc actgcacaga gacctagcag acttccggat ccagcaccca
     3481 gacttgatcc tgctacagta cgtggatgac ttactgctgg ccgccacttc tgagctagac
     3541 tgccaacaag gtactcgggc cctgttacaa accctaggga acctcgggta tcgggcctcg
     3601 gccaagaaag cccaaatttg ccagaaacag gtcaagtatc tggggtatct tctaaaagag
     3661 ggtcagagat ggctgactga ggccagaaaa gagactgtga tggggcagcc tactccgaag
     3721 acccctcgac aactaaggga gttcctaggg aaggcaggct tctgtcgcct cttcatccct
     3781 gggtttgcag aaatggcagc ccccctgtac cctctcacca aaccggggac tctgtttaat
     3841 tggggcccag accaacaaaa ggcctatcaa gaaatcaagc aagctcttct aactgcccca
     3901 gccctggggt tgccagattt gactaagccc tttgaactct ttgtcgacga gaagcagggc
     3961 tacgccaaag gtgtcctaac gcaaaaactg ggaccttggc gtcggccggt ggcctacctg
     4021 tccaaaaagc tagacccagt agcagctggg tggccccctt gcctacggat ggtagcagcc
     4081 attgccgtac tgacaaagga tgcaggcaag ctaaccatgg gacagccact agtcattctg
     4141 gccccccatg cagtagaggc actagtcaaa caaccccccg accgctggct ttccaacgcc
     4201 cggatgactc actatcaggc cttgcttttg gacacggacc gggtccagtt cggaccggtg
     4261 gtagccctga acccgagcgg aggcagtaaa agaacagcag acgggagtga gtttgagccc
     4321 aagaaaaaga gaaaggtgta agaattcgat aatcaacctc tggattacaa aatttgtgaa
     4381 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     4441 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     4501 tcctggttag ttcttgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     4561 ggggctcggc tgttgggcac tgacaattcc gtggtgcgag aattccagac atgataagat
     4621 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     4681 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaagc
     4741 ttgagggcct atttcccatg attccttcat atttgcatat acgatacaag gctgttagag
     4801 agataattag aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta
     4861 gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc
     4921 atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag
     4981 gacgaaacac cgcgggctgg agctgttcgc gcgttttaga gctagaaata gcaagttaaa
     5041 ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgcaa gatggcagcg
     5101 cgaacagctc cagttttttt ttctcgagaa cgctgacgtc atcaacccgc tccaaggaat
     5161 cgcgggccca gtgtcactag gcgggaacac ccagcgcgcg tgcgccctgg caggaagatg
     5221 gctgtgaggg acaggggagt ggcgccctgc aatatttgca tgtcgctatg tgttctggga
     5281 aatcaccata aacgtgaaat gtctttggat ttgggaatct tataagttct gtatgagacc
     5341 actctttccc agtcgtctgc aacctgcaag agttttagag ctagaaatag caagttaaaa
     5401 taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt tttctcgagc
     5461 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     5521 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     5581 gcgagcgagc gcgcagcctt aattaaccta attcactggc cgtcgtttta caacgtcgtg
     5641 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     5701 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     5761 atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     5821 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     5881 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     5941 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     6001 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     6061 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     6121 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     6181 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
     6241 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     6301 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     6361 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     6421 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     6481 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     6541 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     6601 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     6661 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     6721 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     6781 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     6841 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     6901 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     6961 ccggcaacaa ttaatagact ggatggaggc ggataaag
//
