LOCUS       sequence-324011-        5981 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the N-terminal split-intein fragment of PE2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5981)
  AUTHORS   Liu P, Liang SQ, Zheng C, Mintzer E, Zhao YG, Ponnienselvan K, Mir 
            A, Sontheimer EJ, Gao G, Flotte TR, Wolfe SA, Xue W
  TITLE     Improved prime editors enable pathogenic allele correction and 
            cancer modelling in adult mice.
  JOURNAL   Nat Commun. 2021 Apr 9;12(1):2121. doi: 10.1038/s41467-021-22295-w.
  PUBMED    33837189
REFERENCE   2  (bases 1 to 5981)
  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: "2021-04-9"; volume: "12(1):2121. doi"; pages: " 
            10.1038/s41467-021-22295-w"
COMMENT     Sequence Label: pAAV-PE2-N
FEATURES             Location/Qualifiers
     source          1..5981
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   49..178
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      570..579
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             585..611
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             612..2327
                     /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"
     primer_bind     complement(3081..3098)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3087..3294
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3303..3432
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      3507..3962
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3594..3613)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3804..3825
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3979..3998)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4098..4120
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4158..4176)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4244..4348
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4349..4417,4418..5209)
                     /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(4567..4586)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5380..5968
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5869..5888
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtcc tgcaggcagc
       61 tgcgcgctcg ctcgctcact gaggccgccc gggcgtcggg cgacctttgg tcgcccggcc
      121 tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc
      181 ggcctctaga ggtaccgcta gcccgggtcg actagtgatc agtgtgaggg agtgtaaagc
      241 tggtttaaac gttgttgttt ggggttgaat tactcttcta gaccgcggcg cgcctccatg
      301 gatatcaagc ttatggaggc ggtactatgt agatgagaat tcaggagcaa actgggaaaa
      361 gcaactgctt ccaaatattt gtgattttta cagtgtagtt ttggaaaaac tcttagccta
      421 ccaattcttc taagtgtttt aaaatgtggg agccagtaca catgaagtta tagagtgttt
      481 taatgaggct taaatattta ccgtaactat gaaatgctac gcatatcatg ctgttcaggc
      541 tccgtggcca cgcaactcat actaccggtg ccaccatggc tagccccgcc gccaagcgcg
      601 tgaagctgga cgacaagaag tacagcatcg gcctggacat cggcaccaac tctgtgggct
      661 gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca
      721 ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac agcggcgaaa
      781 cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga cggaagaacc
      841 ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac gacagcttct
      901 tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag cggcacccca
      961 tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc atctaccacc
     1021 tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc tatctggccc
     1081 tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg aaccccgaca
     1141 acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag ctgttcgagg
     1201 aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc agactgagca
     1261 agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt
     1321 tcggaaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc
     1381 tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc
     1441 tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg
     1501 acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga
     1561 gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc
     1621 tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg
     1681 gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag ttcatcaagc
     1741 ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc
     1801 tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag
     1861 agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg
     1921 aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg
     1981 gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact
     2041 tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg atgaccaact
     2101 tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact
     2161 tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg
     2221 ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag accaaccgga
     2281 aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg
     2341 tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc
     2401 tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag
     2461 atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa
     2521 cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg
     2581 gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga
     2641 caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc
     2701 acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtgc ctgtcctacg
     2761 agacagagat cctgacagtg gagtatggcc tgctgccaat cggcaagatc gtggagaaga
     2821 ggatcgagtg taccgtgtac tctgtggata acaatggcaa catctataca cagcccgtgg
     2881 cacagtggca cgatagggga gagcaggagg tgttcgagta ttgcctggag gacggcagcc
     2941 tgatcagggc aaccaaggac cacaagttca tgacagtgga tggccagatg ctgcccatcg
     3001 acgagatttt cgagcgggag ctggacctga tgagagtgga taacctgcct aattgagaat
     3061 tcctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     3121 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3181 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     3241 ggggcaggac agcaaggggg aggattggga agagaatagc aggcatgctg gggagcggcc
     3301 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     3361 ggccgggcga ccaaaggtcg cccgacgccc gggcggcctc agtgagcgag cgagcgcgca
     3421 gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     3481 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
     3541 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt
     3601 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3661 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3721 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3781 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc
     3841 tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct attggttaaa
     3901 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     3961 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     4021 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     4081 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     4141 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     4201 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4261 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4321 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4381 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4441 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4501 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4561 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     4621 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4681 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4741 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     4801 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4861 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     4921 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4981 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5041 taaatctgga gccggtgagc gtggaagccg cggtatcatt gcagcactgg ggccagatgg
     5101 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5161 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5221 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5281 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5341 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5401 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5461 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5521 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5581 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5641 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5701 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5761 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5821 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5881 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5941 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac g
//
