LOCUS       sequence-437221-        6483 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV genome expressing ZFcharm targeting Prnp; double perfect match 
            self-silencing binding site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6483)
  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 6483)
  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: pTMB400_ZFcharm_Prnp_DPM
FEATURES             Location/Qualifiers
     source          1..6483
                     /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      432..441
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1563..1583
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2214..2234
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2283..2309
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    2331..2919
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2384..2404)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2802..2813)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2949..2997
                     /note="synthetic polyadenylation signal"
     repeat_region   3746..3886
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3746..3875
                     /label=AAV2 ITR
     rep_origin      3961..4416
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4048..4067)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4258..4279
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4433..4452)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4552..4574
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4612..4630)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4698..4802
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4803..4871,4872..5663)
                     /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(5021..5040)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5834..6422
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6323..6342
                     /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 ctgggctcta 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 acagaggtgt cgtgagctgg atccggatca gcagaccgat
      421 tctgggccag cgccaccatg gccaggacca aacaaactgc cagaaagagc accgggggga
      481 aagcgccccg aaaacagctg gcgaccaaag cagcccgcaa aagcgcaccg ggaggtgcaa
      541 gcagtggggc aggaagttcc tcaggaggaa gtgcggcggg atccggaagc ggtggcgcat
      601 caagtggcgc cgcaagtagc agtgccgggt ccgcagctgg ctccgggtcc tcccctgttg
      661 agatctacaa gaccgtgtcc gcctggaagc ggcagccaat cagagtgctg agcctgttcg
      721 gcaatattga caaggaactg aagtctctgg gcttcctgga aagcggcagc cacagcgagg
      781 gcggcacact gaagtacgtg gaagatgtga ccaacgtggt gcggagagat gtggaaaagt
      841 ggggaccttt cgacctgctg tacggcagca cccagcctct gggcggaagc tgcgacaggt
      901 gtcctggctg gtacatgttc caatttcata gaatcctgca gtacgccaga cctagacagg
      961 agtctcagcc cttcttctgg atcttcatgg ataacctgct gctcacagag gacgaccaga
     1021 tgaccaccgt gcggttcctg caaacagagg ccgtcaccct gcgcgacgtg agatctagag
     1081 tgctgcagaa cgccgtccgg gtgtggagca acatccccgg tcttaaaagc aagcacgagg
     1141 ctctgtcccc aaaagaggaa gagagcctgc agggccacgt gcggaccaga tctaagctgg
     1201 ccgctcctaa agtggacccc ctggtgaaga actgcctgct gcctctgaga gagtacttta
     1261 agtatttcag ccagagcagc ctgcccctgg gaggacctag cagcggcgcc ccacctccaa
     1321 gcggaggctc tccagctggc agccctacca gcacagagga aggcacaagc gagagcgcca
     1381 cacctgagag cggccctggc acctccaccg agcccagcga gggctctgcc cctggcagcc
     1441 ccgccggaag ccccacctct acagaagaag gcaccagcac agagccttct gaaggctctg
     1501 cccctggaac atccaccgag cctagcgaga agcggactgc cgatggatct gagtttgaga
     1561 gccctaagaa aaaaagaaag gttggagtgc ctgctgctat ggccgagaga ccatttcagt
     1621 gcagaatctg catgcgcaat ttcagcacca gcggaaatct cacaagacac atccggaccc
     1681 ataccggcga gaagcccttt gcctgcgata tctgcggcag aaagttcgct ctgtctcagg
     1741 acctgaaccg gcacaccaag atccacacac accccagagc ccctatcccc aagcctttcc
     1801 agtgccggat ttgcatgcgg aacttcagcg acaggtccgc cctgagcaga catatcagaa
     1861 cacacaccgg cgaaaagcct ttcgcctgtg atatctgtgg cagaaagttc gcccggaacg
     1921 acgacagaaa aaagcacaca aagatccaca ccggatctca gaaacctttc caatgtagaa
     1981 tctgcatgag aaacttcagc cagagcggca acctggccag acacatccgg acccacacag
     2041 gcgaaaagcc tttcgcctgt gatatctgcg gccggaaatt cgcccagaga atcgacctga
     2101 cccggcacac caagatccac ctgagacaga aggacgccgc cagaggcagc ggcggcgacg
     2161 ccaagagcct gaccgcctgg tccaagagga ccgccgacgg cagcgagttc gagcccaaga
     2221 aaaagcggaa ggtgtccggc tctgaaaccc ctggcacaag cgagtctgct acacctgagt
     2281 cctatcccta tgacgtgccc gattatgcct gatagaattc gcttatcgat aatcaacctc
     2341 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     2401 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     2461 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     2521 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     2581 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     2641 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     2701 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctatgttg
     2761 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     2821 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     2881 ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccga gcgctgctcg
     2941 agagatctaa taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtggcg
     3001 gccgctagtg gcagcaccag ctcagggctg gaggtggtgg ctcctgaggg tacctcagcc
     3061 ccaggtggtg gcccgggaac cctggatgac agtgccacca tttgccgtgt ctgccagaag
     3121 ccaggcgatc tggttatgtg caaccagtgt gagttttgtt tccacctgga ctgtcacctg
     3181 ccggccctgc aggatgtacc aggggaggag tggagctgct cactctgcca tgtgctccct
     3241 gacctgaagg aggaggatgg cagcctcagc ctggatggtg cagacagcac tggcgtggtg
     3301 gccaagctct caccagccaa ccagcggaaa tgtgagcgtg tactgctggc cctattctgt
     3361 cacgaaccct gccgccccct gcatcagctg gctaccgact ccaccttctc cctggaccag
     3421 cccggtggca ccctggatct gaccctgatc cgtgcccgcc tccaggagaa gttgtcacct
     3481 ccctacagct ccccacagga gtttgcccag gatgtgggcc gcatgttcaa gcaattcaac
     3541 aagttaactg aggacaaggc agacgtgcag tccatcatcg gcctgcagcg cttcttcgag
     3601 acgcgcatga acgaggcctt cggtgacacc aagttctctg ctgtgctggt ggagcccccg
     3661 ccgatgagcc tgcctggtgc tggcctgagt tcccaggagc tgtctggtgg ccctggtgat
     3721 ggcccctgcc caactttaag cttgcaggaa cccctagtga tggagttggc cactccctct
     3781 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     3841 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     3901 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     3961 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4021 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4081 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4141 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     4201 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4261 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     4321 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     4381 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     4441 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     4501 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     4561 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     4621 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     4681 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4741 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4801 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4861 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4921 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4981 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5041 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5101 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5161 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5221 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5281 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5341 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5401 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5461 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtggaa
     5521 gccgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5581 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5641 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5701 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5761 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5821 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5881 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5941 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6001 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6061 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6121 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6181 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6241 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6301 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6361 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6421 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6481 tgt
//
