LOCUS       sequence-437218-        6443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV genome expressing ZFcharm targeting Prnp; single mismatch 
            self-silencing binding site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6443)
  AUTHORS   Neumann EN, Bertozzi TM, Wu E, Serack F, Harvey JW, Brauer PP, 
            Pirtle CP, Coffey A, Howard M, Kamath N, Lenz K, Guzman K, Raymond 
            MH, Khalil AS, Deverman BE, Minikel EV, Vallabh SM, Weissman JS
  TITLE     Brainwide silencing of prion protein by AAV-mediated delivery of an 
            engineered compact epigenetic editor.
  JOURNAL   Science. 2024 Jun 28;384(6703):ado7082. doi: 
            10.1126/science.ado7082. Epub 2024 Jun 28.
  PUBMED    38935715
REFERENCE   2  (bases 1 to 6443)
  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: "Science."; 
            date: "2024-06-28"; volume: "384(6703):ado7082. doi"; pages: " 
            10.1126/science.ado7082. Epub 2024 Jun 28"
COMMENT     Sequence Label: pTMB398_ZFcharm_Prnp_SMM
FEATURES             Location/Qualifiers
     source          1..6443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        173..384
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      392..401
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1523..1543
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2174..2194
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2243..2269
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    2291..2879
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2344..2364)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2762..2773)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2909..2957
                     /note="synthetic polyadenylation signal"
     repeat_region   3706..3846
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3706..3835
                     /label=AAV2 ITR
     rep_origin      3921..4376
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4008..4027)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4218..4239
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4393..4412)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4512..4534
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4572..4590)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4658..4762
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4763..4831,4832..5623)
                     /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(4981..5000)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5794..6382
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6283..6302
                     /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 gcggcctact agtggatcag cagaccgatt gtgggctcta gagggcagag
      181 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aaccggtgcc
      241 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt
      301 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc
      361 aacgggtttg ccgccagaac acagcctcag cgccaccatg gccaggacca aacaaactgc
      421 cagaaagagc accgggggga aagcgccccg aaaacagctg gcgaccaaag cagcccgcaa
      481 aagcgcaccg ggaggtgcaa gcagtggggc aggaagttcc tcaggaggaa gtgcggcggg
      541 atccggaagc ggtggcgcat caagtggcgc cgcaagtagc agtgccgggt ccgcagctgg
      601 ctccgggtcc tcccctgttg agatctacaa gaccgtgtcc gcctggaagc ggcagccaat
      661 cagagtgctg agcctgttcg gcaatattga caaggaactg aagtctctgg gcttcctgga
      721 aagcggcagc cacagcgagg gcggcacact gaagtacgtg gaagatgtga ccaacgtggt
      781 gcggagagat gtggaaaagt ggggaccttt cgacctgctg tacggcagca cccagcctct
      841 gggcggaagc tgcgacaggt gtcctggctg gtacatgttc caatttcata gaatcctgca
      901 gtacgccaga cctagacagg agtctcagcc cttcttctgg atcttcatgg ataacctgct
      961 gctcacagag gacgaccaga tgaccaccgt gcggttcctg caaacagagg ccgtcaccct
     1021 gcgcgacgtg agatctagag tgctgcagaa cgccgtccgg gtgtggagca acatccccgg
     1081 tcttaaaagc aagcacgagg ctctgtcccc aaaagaggaa gagagcctgc agggccacgt
     1141 gcggaccaga tctaagctgg ccgctcctaa agtggacccc ctggtgaaga actgcctgct
     1201 gcctctgaga gagtacttta agtatttcag ccagagcagc ctgcccctgg gaggacctag
     1261 cagcggcgcc ccacctccaa gcggaggctc tccagctggc agccctacca gcacagagga
     1321 aggcacaagc gagagcgcca cacctgagag cggccctggc acctccaccg agcccagcga
     1381 gggctctgcc cctggcagcc ccgccggaag ccccacctct acagaagaag gcaccagcac
     1441 agagccttct gaaggctctg cccctggaac atccaccgag cctagcgaga agcggactgc
     1501 cgatggatct gagtttgaga gccctaagaa aaaaagaaag gttggagtgc ctgctgctat
     1561 ggccgagaga ccatttcagt gcagaatctg catgcgcaat ttcagcacca gcggaaatct
     1621 cacaagacac atccggaccc ataccggcga gaagcccttt gcctgcgata tctgcggcag
     1681 aaagttcgct ctgtctcagg acctgaaccg gcacaccaag atccacacac accccagagc
     1741 ccctatcccc aagcctttcc agtgccggat ttgcatgcgg aacttcagcg acaggtccgc
     1801 cctgagcaga catatcagaa cacacaccgg cgaaaagcct ttcgcctgtg atatctgtgg
     1861 cagaaagttc gcccggaacg acgacagaaa aaagcacaca aagatccaca ccggatctca
     1921 gaaacctttc caatgtagaa tctgcatgag aaacttcagc cagagcggca acctggccag
     1981 acacatccgg acccacacag gcgaaaagcc tttcgcctgt gatatctgcg gccggaaatt
     2041 cgcccagaga atcgacctga cccggcacac caagatccac ctgagacaga aggacgccgc
     2101 cagaggcagc ggcggcgacg ccaagagcct gaccgcctgg tccaagagga ccgccgacgg
     2161 cagcgagttc gagcccaaga aaaagcggaa ggtgtccggc tctgaaaccc ctggcacaag
     2221 cgagtctgct acacctgagt cctatcccta tgacgtgccc gattatgcct gatagaattc
     2281 gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     2341 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     2401 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     2461 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     2521 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     2581 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     2641 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     2701 ttggctgctc gcctatgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     2761 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     2821 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca
     2881 tcgataccga gcgctgctcg agagatctaa taaaatatct ttattttcat tacatctgtg
     2941 tgttggtttt ttgtgtggcg gccgctagtg gcagcaccag ctcagggctg gaggtggtgg
     3001 ctcctgaggg tacctcagcc ccaggtggtg gcccgggaac cctggatgac agtgccacca
     3061 tttgccgtgt ctgccagaag ccaggcgatc tggttatgtg caaccagtgt gagttttgtt
     3121 tccacctgga ctgtcacctg ccggccctgc aggatgtacc aggggaggag tggagctgct
     3181 cactctgcca tgtgctccct gacctgaagg aggaggatgg cagcctcagc ctggatggtg
     3241 cagacagcac tggcgtggtg gccaagctct caccagccaa ccagcggaaa tgtgagcgtg
     3301 tactgctggc cctattctgt cacgaaccct gccgccccct gcatcagctg gctaccgact
     3361 ccaccttctc cctggaccag cccggtggca ccctggatct gaccctgatc cgtgcccgcc
     3421 tccaggagaa gttgtcacct ccctacagct ccccacagga gtttgcccag gatgtgggcc
     3481 gcatgttcaa gcaattcaac aagttaactg aggacaaggc agacgtgcag tccatcatcg
     3541 gcctgcagcg cttcttcgag acgcgcatga acgaggcctt cggtgacacc aagttctctg
     3601 ctgtgctggt ggagcccccg ccgatgagcc tgcctggtgc tggcctgagt tcccaggagc
     3661 tgtctggtgg ccctggtgat ggcccctgcc caactttaag cttgcaggaa cccctagtga
     3721 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     3781 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     3841 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     3901 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3961 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     4021 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4081 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     4141 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4201 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     4261 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     4321 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     4381 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     4441 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     4501 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     4561 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     4621 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4681 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4741 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4801 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4861 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4921 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4981 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     5041 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5101 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5161 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     5221 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5281 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     5341 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5401 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5461 tggagccggt gagcgtggaa gccgcggtat cattgcagca ctggggccag atggtaagcc
     5521 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5581 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     5641 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5701 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5761 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5821 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5881 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5941 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6001 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6061 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6121 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6181 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6241 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6301 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6361 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6421 ttgctggcct tttgctcaca tgt
//
