LOCUS       sequence-369050-        7489 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  p3-PE2-N-term(1152) in AAV2 backbone (ITRs).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7489)
  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 7489)
  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(1152)-N-term
FEATURES             Location/Qualifiers
     source          1..7489
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      390..978
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     879..898
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    1266..1287
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1302..1307,1308..1325,1326..1332)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1340..1356
                     /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     1345..1367
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1364..1380
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1364..1380
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   1407..1536
                     /label=AAV2 ITR
     CDS             1917..1937
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1938..3653
                     /codon_start=1
                     /product="N-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(N)
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
     CDS             5787..5807
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5826..5947
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5837..5856)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5891..5910
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   5956..6085
                     /label=AAV2 ITR
     primer_bind     complement(6104..6121)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6104..6120)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6113..6135)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6262..6717
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6349..6368)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6559..6580
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6743..6847
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6848..6916,6917..7489,1..219)
                     /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(7066..7085)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
       61 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
      121 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
      181 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
      241 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
      301 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
      361 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
      421 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
      481 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
      541 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
      601 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
      661 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
      721 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
      781 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
      841 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
      901 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
      961 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     1021 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     1081 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     1141 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     1201 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     1261 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     1321 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     1381 catgattacg ccagatttaa ttaaggctgc gcgctcgctc gctcactgag gccgcccggg
     1441 caaagcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca
     1501 gagagggagt ggccaactcc atcactaggg gttcctgcgg cctctagatc agaccggcgc
     1561 gccgggggag gctgctggtg aatattaacc aaggtcaccc cagttatcgg aggagcaaac
     1621 aggggctaag tccacacgcg tggtaccgtc tgtctgcaca tttcgtagag cgagtgttcc
     1681 gatactctaa tctccctagg caaggttcat atttgtgtag gttacttatt ctccttttgt
     1741 tgactaagtc aataatcaga atcagcaggt ttggagtcag cttggcaggg atcagcagcc
     1801 tgggttggaa ggagggggta taaaagcccc ttcaccagga gaagccgtca cacagatcca
     1861 caagctcctg ggccgccacc atgaaacgga cagccgatgg atctgagttt gagtctccga
     1921 agaaaaaacg caaggtcgac aagaagtaca gcatcggcct ggacatcggc accaactctg
     1981 tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg
     2041 gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg
     2101 gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga
     2161 agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca
     2221 gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc
     2281 accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct
     2341 accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc
     2401 tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc
     2461 ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt
     2521 tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac
     2581 tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg
     2641 gcctgttcgg aaacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact
     2701 tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg
     2761 acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc
     2821 tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc
     2881 ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga
     2941 aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca
     3001 agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca
     3061 tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag
     3121 aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc
     3181 tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca
     3241 accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg
     3301 ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct
     3361 ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga
     3421 ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg
     3481 agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa
     3541 agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca
     3601 accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg
     3661 actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg
     3721 atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc
     3781 tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc
     3841 tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat
     3901 acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg
     3961 gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc
     4021 tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc
     4081 agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg
     4141 gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc
     4201 ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga
     4261 acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc
     4321 tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc
     4381 tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact
     4441 acgatgtgga cgctatcgtg cctcagagct ttctgaagga cgactccatc gacaacaagg
     4501 tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg
     4561 tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt acccagagaa
     4621 agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct
     4681 tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg
     4741 actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa gtgaaagtga
     4801 tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc
     4861 gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg
     4921 ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt
     4981 acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt
     5041 acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg
     5101 agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata
     5161 agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga
     5221 aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca
     5281 gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca
     5341 gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagggtgggg
     5401 gaggttctgg agggggaggg tcaggtggcg gaggatcttg tcttagctac gagacagaga
     5461 ttcttacagt ggagtatgga cttcttccca tcggtaaaat tgttgaaaag aggatcgagt
     5521 gcacggtcta cagtgtcgat aataatggta acatatatac gcaacccgtc gcgcagtggc
     5581 atgatagagg cgagcaggaa gtatttgagt attgtctgga ggatggatct ctgattcgcg
     5641 cgactaagga ccataagttt atgacggttg acggacaaat gctcccaata gacgagatat
     5701 ttgaacgaga actggatctt atgagagtcg ataacttgcc gaatagcgga ggcagtaaaa
     5761 gaacagcaga cgggagtgag tttgagccca agaaaaagag aaaggtgtaa gaattccaga
     5821 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     5881 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     5941 acaagttgcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     6001 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     6061 cctcagtgag cgagcgagcg cgcagcctta attaacctaa ttcactggcc gtcgttttac
     6121 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     6181 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     6241 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     6301 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     6361 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     6421 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     6481 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     6541 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     6601 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     6661 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     6721 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     6781 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     6841 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     6901 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     6961 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     7021 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     7081 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     7141 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     7201 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     7261 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     7321 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     7381 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     7441 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagtt
//
