LOCUS       sequence-350105-        6993 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminus of PE2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6993)
  AUTHORS   Jang H, Jo DH, Cho CS, Shin JH, Seo JH, Yu G, Gopalappa R, Kim D, 
            Cho SR, Kim JH, Kim HH
  TITLE     Application of prime editing to the correction of mutations and 
            phenotypes in adult mice with liver and eye diseases.
  JOURNAL   Nat Biomed Eng. 2021 Aug 26. pii: 10.1038/s41551-021-00788-9. doi: 
            10.1038/s41551-021-00788-9.
  PUBMED    34446856
REFERENCE   2  (bases 1 to 6993)
  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 Biomed 
            Eng. 2021 Aug 26. pii: 10.1038/s41551-021-00788-9. doi: 
            10.1038/s41551-021-00788-9."
COMMENT     Sequence Label: TS-PE2-C-WPRE
FEATURES             Location/Qualifiers
     source          1..6993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   155..284
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             3548..3568
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    3588..4176
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3641..3661)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4059..4070)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4177..4401
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4410..4550
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4410..4539
                     /label=AAV2 ITR
     rep_origin      4625..5080
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4712..4731)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4922..4943
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5097..5116)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5216..5238
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5276..5294)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5362..5466
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5467..5535,5536..6327)
                     /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(5685..5704)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6498..6993,1..93)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     join(6987..6993,1..13)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
       61 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
      121 ttcctggcct tttgctggcc ttttgctcac atgtcctgca ggcagctgcg cgctcgctcg
      181 ctcactgagg ccgcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg
      241 agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctgcggcc tctagacttg
      301 tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac tgacatccac
      361 tttgcctttc tctccacagt tcttctacag caacatcatg aactttttca agaccgagat
      421 taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg
      481 ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc
      541 ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat
      601 cctgcccaag aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa
      661 gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga
      721 aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga
      781 aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt
      841 gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg
      901 gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc
      961 caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga
     1021 ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat
     1081 cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt
     1141 gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat
     1201 ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac
     1261 catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca
     1321 gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag gtgactctgg
     1381 aggatctagc ggaggatcct ctggcagcga gacaccagga acaagcgagt cagcaacacc
     1441 agagagcagt ggcggcagca gcggcggcag cagcacccta aatatagaag atgagtatcg
     1501 gctacatgag acctcaaaag agccagatgt ttctctaggg tccacatggc tgtctgattt
     1561 tcctcaggcc tgggcggaaa ccgggggcat gggactggca gttcgccaag ctcctctgat
     1621 catacctctg aaagcaacct ctacccccgt gtccataaaa caatacccca tgtcacaaga
     1681 agccagactg gggatcaagc cccacataca gagactgttg gaccagggaa tactggtacc
     1741 ctgccagtcc ccctggaaca cgcccctgct acccgttaag aaaccaggga ctaatgatta
     1801 taggcctgtc caggatctga gagaagtcaa caagcgggtg gaagacatcc accccaccgt
     1861 gcccaaccct tacaacctct tgagcgggct cccaccgtcc caccagtggt acactgtgct
     1921 tgatttaaag gatgcctttt tctgcctgag actccacccc accagtcagc ctctcttcgc
     1981 ctttgagtgg agagatccag agatgggaat ctcaggacaa ttgacctgga ccagactccc
     2041 acagggtttc aaaaacagtc ccaccctgtt taatgaggca ctgcacagag acctagcaga
     2101 cttccggatc cagcacccag acttgatcct gctacagtac gtggatgact tactgctggc
     2161 cgccacttct gagctagact gccaacaagg tactcgggcc ctgttacaaa ccctagggaa
     2221 cctcgggtat cgggcctcgg ccaagaaagc ccaaatttgc cagaaacagg tcaagtatct
     2281 ggggtatctt ctaaaagagg gtcagagatg gctgactgag gccagaaaag agactgtgat
     2341 ggggcagcct actccgaaga cccctcgaca actaagggag ttcctaggga aggcaggctt
     2401 ctgtcgcctc ttcatccctg ggtttgcaga aatggcagcc cccctgtacc ctctcaccaa
     2461 accggggact ctgtttaatt ggggcccaga ccaacaaaag gcctatcaag aaatcaagca
     2521 agctcttcta actgccccag ccctggggtt gccagatttg actaagccct ttgaactctt
     2581 tgtcgacgag aagcagggct acgccaaagg tgtcctaacg caaaaactgg gaccttggcg
     2641 tcggccggtg gcctacctgt ccaaaaagct agacccagta gcagctgggt ggcccccttg
     2701 cctacggatg gtagcagcca ttgccgtact gacaaaggat gcaggcaagc taaccatggg
     2761 acagccacta gtcattctgg ccccccatgc agtagaggca ctagtcaaac aaccccccga
     2821 ccgctggctt tccaacgccc ggatgactca ctatcaggcc ttgcttttgg acacggaccg
     2881 ggtccagttc ggaccggtgg tagccctgaa cccggctacg ctgctcccac tgcctgagga
     2941 agggctgcaa cacaactgcc ttgatatcct ggccgaagcc cacggaaccc gacccgacct
     3001 aacggaccag ccgctcccag acgccgacca cacctggtac acggatggaa gcagtctctt
     3061 acaagaggga cagcgtaagg cgggagctgc ggtgaccacc gagaccgagg taatctgggc
     3121 taaagccctg ccagccggga catccgctca gcgggctgaa ctgatagcac tcacccaggc
     3181 cctaaagatg gcagaaggta agaagctaaa tgtttatact gatagccgtt atgcttttgc
     3241 tactgcccat atccatggag aaatatacag aaggcgtggg tggctcacat cagaaggcaa
     3301 agagatcaaa aataaagacg agatcttggc cctactaaaa gccctctttc tgcccaaaag
     3361 acttagcata atccattgtc caggacatca aaagggacac agcgccgagg ctagaggcaa
     3421 ccggatggct gaccaagcgg cccgaaaggc agccatcaca gagactccag acacctctac
     3481 cctcctcata gaaaattcat caccctctgg cggctcaaaa agaaccgccg acggcagcga
     3541 attcgagccc aagaagaaga ggaaagtcta accggtcatc atcaccaaat caacctctgg
     3601 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     3661 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     3721 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     3781 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     3841 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     3901 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     3961 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca
     4021 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     4081 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     4141 agacgagtcg gatctccctt tgggccgcct ccccgcctgt gccttctagt tgccagccat
     4201 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     4261 tttcctaata aaatgagaaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     4321 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     4381 gggatgcggt gggctctatg ggcggccgca ggaaccccta gtgatggagt tggccactcc
     4441 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     4501 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat
     4561 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca
     4621 tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4681 accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc
     4741 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4801 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt
     4861 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4921 agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat
     4981 ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa
     5041 tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca
     5101 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     5161 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     5221 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     5281 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     5341 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5401 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5461 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     5521 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     5581 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     5641 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     5701 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     5761 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5821 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5881 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5941 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     6001 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     6061 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     6121 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     6181 ggaagccgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     6241 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     6301 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     6361 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6421 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6481 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6541 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6601 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     6661 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6721 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6781 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6841 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6901 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6961 ggagagcgca cgagggagct tccaggggga aac
//
