LOCUS       sequence-365390-        7559 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Deliver a split compact Prime Editor in vivo.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7559)
  AUTHORS   Zheng C, Liang SQ, Liu B, Liu P, Kwan SY, Wolfe SA, Xue W
  TITLE     A flexible split prime editor using truncated reverse transcriptase 
            improves dual-AAV delivery in mouse liver.
  JOURNAL   Mol Ther. 2022 Mar 2;30(3):1343-1351. doi: 
            10.1016/j.ymthe.2022.01.005. Epub 2022 Jan 5.
  PUBMED    34998953
REFERENCE   2  (bases 1 to 7559)
  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"; doi: 
            "10.1016/j.ymthe.2022.01.005"; journalName: "Mol Ther"; date: 
            "2022-03-2- 2"; volume: "30"; issue: "3"; pages: "1343-1351"
COMMENT     Sequence Label: AAV8-cPE-C
FEATURES             Location/Qualifiers
     source          1..7559
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             514..2898
                     /codon_start=1
                     /product="C-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(C)
                     /translation="CFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILED
                     IVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGK
                     TILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKG
                     ILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQI
                     LKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDN
                     KVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDK
                     AGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQF
                     YKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGK
                     ATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMP
                     QVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKV
                     EKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELEN
                     GRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLD
                     EIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKY
                     FDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD"
     CDS             4531..4551
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4560..4577
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4600..4617)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4606..4813
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4822..4962
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4822..4951
                     /label=AAV2 ITR
     rep_origin      5037..5492
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5124..5143)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5334..5355
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5509..5528)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5628..5650
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5688..5706)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5774..5878
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5879..5947,5948..6739)
                     /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(6097..6116)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6910..7498
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7399..7418
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggcctcta gaatggaggc ggtactatgt agatgagaat tcaggagcaa
      181 actgggaaaa gcaactgctt ccaaatattt gtgattttta cagtgtagtt ttggaaaaac
      241 tcttagccta ccaattcttc taagtgtttt aaaatgtggg agccagtaca catgaagtta
      301 tagagtgttt taatgaggct taaatattta ccgtaactat gaaatgctac gcatatcatg
      361 ctgttcaggc tccgtggcca cgcaactcat actaccggtg ccaccatgat caagattgct
      421 acacggaaat acctgggaaa gcagaacgtg tacgacatcg gcgtggagcg ggatcacaac
      481 ttcgccctga agaatggctt tatcgccagc aattgcttcg actccgtgga aatctccggc
      541 gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag
      601 gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc
      661 ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg
      721 ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg
      781 agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc
      841 ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg
      901 acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag
      961 cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag
     1021 gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa
     1081 atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag
     1141 cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa
     1201 aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg
     1261 tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga cgctatcgtg
     1321 cctcagagct ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag
     1381 aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac
     1441 tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag
     1501 gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg
     1561 gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag
     1621 tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg
     1681 gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac
     1741 cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct
     1801 aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc
     1861 gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc
     1921 atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg
     1981 atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc
     2041 gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca
     2101 ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga
     2161 aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct
     2221 gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag
     2281 ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg
     2341 gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc
     2401 ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag
     2461 ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat
     2521 gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac
     2581 aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg
     2641 gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc
     2701 agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc
     2761 gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg
     2821 ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg
     2881 tctcagctgg gaggtgactc tggaggatct agcggaggat cctctggcag cgagacacca
     2941 ggaacaagcg agtcagcaac accagagagc agtggcggca gcagcggcgg cagcagcacc
     3001 ctaaatatag aagatgagta tcggctacat gagacctcaa aagagccaga tgtttctcta
     3061 gggtccacat ggctgtctga ttttcctcag gcctgggcgg aaaccggggg catgggactg
     3121 gcagttcgcc aagctcctct gatcatacct ctgaaagcaa cctctacccc cgtgtccata
     3181 aaacaatacc ccatgtcaca agaagccaga ctggggatca agccccacat acagagactg
     3241 ttggaccagg gaatactggt accctgccag tccccctgga acacgcccct gctacccgtt
     3301 aagaaaccag ggactaatga ttataggcct gtccaggatc tgagagaagt caacaagcgg
     3361 gtggaagaca tccaccccac cgtgcccaac ccttacaacc tcttgagcgg gctcccaccg
     3421 tcccaccagt ggtacactgt gcttgattta aaggatgcct ttttctgcct gagactccac
     3481 cccaccagtc agcctctctt cgcctttgag tggagagatc cagagatggg aatctcagga
     3541 caattgacct ggaccagact cccacagggt ttcaaaaaca gtcccaccct gtttaatgag
     3601 gcactgcaca gagacctagc agacttccgg atccagcacc cagacttgat cctgctacag
     3661 tacgtggatg acttactgct ggccgccact tctgagctag actgccaaca aggtactcgg
     3721 gccctgttac aaaccctagg gaacctcggg tatcgggcct cggccaagaa agcccaaatt
     3781 tgccagaaac aggtcaagta tctggggtat cttctaaaag agggtcagag atggctgact
     3841 gaggccagaa aagagactgt gatggggcag cctactccga agacccctcg acaactaagg
     3901 gagttcctag ggaaggcagg cttctgtcgc ctcttcatcc ctgggtttgc agaaatggca
     3961 gcccccctgt accctctcac caaaccgggg actctgttta attggggccc agaccaacaa
     4021 aaggcctatc aagaaatcaa gcaagctctt ctaactgccc cagccctggg gttgccagat
     4081 ttgactaagc cctttgaact ctttgtcgac gagaagcagg gctacgccaa aggtgtccta
     4141 acgcaaaaac tgggaccttg gcgtcggccg gtggcctacc tgtccaaaaa gctagaccca
     4201 gtagcagctg ggtggccccc ttgcctacgg atggtagcag ccattgccgt actgacaaag
     4261 gatgcaggca agctaaccat gggacagcca ctagtcattc tggcccccca tgcagtagag
     4321 gcactagtca aacaaccccc cgaccgctgg ctttccaacg cccggatgac tcactatcag
     4381 gccttgcttt tggacacgga ccgggtccag ttcggaccgg tggtagccct gaacccggct
     4441 acgctgctcc cactgcctga ggaagggctg caacacaact gccttgattc tggcggctca
     4501 aaaagaaccg ccgacggcag cgaattcgag cccaagaaga agaggaaagt ctaaccggtc
     4561 atcatcacca tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     4621 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     4681 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     4741 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gagaatagca
     4801 ggcatgctgg ggagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     4861 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     4921 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
     4981 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     5041 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     5101 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     5161 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     5221 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     5281 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     5341 cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg
     5401 gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     5461 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     5521 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     5581 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     5641 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     5701 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     5761 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     5821 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     5881 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     5941 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     6001 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     6061 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     6121 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     6181 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     6241 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     6301 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     6361 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     6421 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     6481 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     6541 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtggaagccg
     6601 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     6661 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     6721 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     6781 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     6841 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     6901 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     6961 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     7021 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     7081 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     7141 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     7201 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     7261 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     7321 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     7381 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     7441 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     7501 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt
//
