LOCUS       sequence-437216-        6678 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV genome expressing ZFcharm Kv1 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 6678)
  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 6678)
  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: pTMB340_ZFcharm_Kv1_Prnp_DPM
FEATURES             Location/Qualifiers
     source          1..6678
                     /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..2468
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEP"
     CDS             2478..2504
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    2526..3114
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2579..2599)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2997..3008)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3144..3192
                     /note="synthetic polyadenylation signal"
     repeat_region   3941..4081
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3941..4070
                     /label=AAV2 ITR
     rep_origin      4156..4611
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4243..4262)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4453..4474
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4628..4647)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4747..4769
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4807..4825)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4893..4997
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4998..5066,5067..5858)
                     /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(5216..5235)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6029..6617
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6518..6537
                     /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 ccagaaccct ggtcaccttc aaagacgtgt tcgtggactt caccagagaa gaatggaagc
     2341 tgctggatac cgctcagcag atcgtgtaca gaaacgtgat gctggaaaac tacaagaatc
     2401 tggtgtccct gggctaccag ctgaccaagc ctgacgtgat cctgagactg gaaaagggcg
     2461 aggaacccgg gagcggctat ccctatgacg tgcccgatta tgcctgatag aattcgctta
     2521 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     2581 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     2641 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     2701 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     2761 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     2821 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     2881 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     2941 tgctcgccta tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     3001 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     3061 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     3121 accgagcgct gctcgagaga tctaataaaa tatctttatt ttcattacat ctgtgtgttg
     3181 gttttttgtg tggcggccgc tagtggcagc accagctcag ggctggaggt ggtggctcct
     3241 gagggtacct cagccccagg tggtggcccg ggaaccctgg atgacagtgc caccatttgc
     3301 cgtgtctgcc agaagccagg cgatctggtt atgtgcaacc agtgtgagtt ttgtttccac
     3361 ctggactgtc acctgccggc cctgcaggat gtaccagggg aggagtggag ctgctcactc
     3421 tgccatgtgc tccctgacct gaaggaggag gatggcagcc tcagcctgga tggtgcagac
     3481 agcactggcg tggtggccaa gctctcacca gccaaccagc ggaaatgtga gcgtgtactg
     3541 ctggccctat tctgtcacga accctgccgc cccctgcatc agctggctac cgactccacc
     3601 ttctccctgg accagcccgg tggcaccctg gatctgaccc tgatccgtgc ccgcctccag
     3661 gagaagttgt cacctcccta cagctcccca caggagtttg cccaggatgt gggccgcatg
     3721 ttcaagcaat tcaacaagtt aactgaggac aaggcagacg tgcagtccat catcggcctg
     3781 cagcgcttct tcgagacgcg catgaacgag gccttcggtg acaccaagtt ctctgctgtg
     3841 ctggtggagc ccccgccgat gagcctgcct ggtgctggcc tgagttccca ggagctgtct
     3901 ggtggccctg gtgatggccc ctgcccaact ttaagcttgc aggaacccct agtgatggag
     3961 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     4021 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     4081 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     4141 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     4201 cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc
     4261 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     4321 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     4381 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     4441 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct
     4501 attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga
     4561 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     4621 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     4681 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     4741 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     4801 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     4861 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4921 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4981 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5041 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5101 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5161 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     5221 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5281 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     5341 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5401 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5461 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5521 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5581 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5641 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5701 ccggtgagcg tggaagccgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5761 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5821 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5881 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5941 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6001 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6061 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6121 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6181 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6241 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6301 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6361 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6421 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6481 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6541 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6601 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6661 ggccttttgc tcacatgt
//
