LOCUS       sequence-469452-        6759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of Venus-P2A-Tau-RING-I18R/M72E from hSyn promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6759)
  AUTHORS   Miller LVC, Papa G, Vaysburd M, Cheng S, Sweeney PW, Smith A, Franco
            C, Katsinelos T, Huang M, Sanford SAI, Benn J, Farnsworth J, 
            Higginson K, Joyner H, McEwan WA, James LC
  TITLE     Co-opting templated aggregation to degrade pathogenic tau assemblies
            and improve motor function.
  JOURNAL   Cell. 2024 Oct 17;187(21):5967-5980.e17. doi: 
            10.1016/j.cell.2024.08.024. Epub 2024 Sep 13.
  PUBMED    39276772
REFERENCE   2  (bases 1 to 6759)
  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"; doi: 
            "10.1016/j.cell.2024.08.024"; journalName: "Cell"; date: 
            "2024-10-17- 17"; volume: "187"; issue: "21"; pages: "5967-5980.e17"
COMMENT     Sequence Label: AAV 9P31-hSyn-Venus-P2A-Tau-RING I18R/M72E
FEATURES             Location/Qualifiers
     source          1..6759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   72..212
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   83..212
                     /label=AAV2 ITR
     polyA_signal    252..728
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     510..529
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    760..1348
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             866..877
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1275..1295
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2810..2866)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(join(2882..3592,3593..3595,3596..3598))
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2924..2945)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3273..3292
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3532..3553
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(3612..4059)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   4104..4233
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(4295..4883)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4375..4394)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5054..5845,5846..5914))
                     /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     5677..5696
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5915..6019)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6087..6105
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6143..6165)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6265..6284
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6301..6756)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6438..6459)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6650..6669
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atggttgctt tgacgtatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
       61 catcaggcgc ccctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa
      121 gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga
      181 gggagtggcc aactccatca ctaggggttc ctgcggccgc tcggtccgca cgtggttacc
      241 tacaaaatca gaaggacagg gaagggagca gtggttcacg cctgtaatcc cagcaatttg
      301 ggaggccaag gtgggtagat cacctgagat taggagttgg agaccagcct ggccaatatg
      361 gtgaaacccc gtctctacca aaaaaacaaa aattagctga gcctggtcat gcatgcctgg
      421 aatcccaaca actcgggagg ctgaggcagg agaatcgctt gaacccagga ggcggagatt
      481 gcagtgagcc aagattgtgc cactgcactc cagcttggtt cccaatagac cccgcaggcc
      541 ctacaggttg tcttcccaac ttgccccttg ctccatacca cccccctcca ccccataata
      601 ttatagaagg acacctagtc agacaaaatg atgcaactta attttattag gacaaggctg
      661 gtgggcactg gagtggcaac ttccagggcc aggagaggca ctggggaggg gtcacaggga
      721 tgccacccgt agatctctcg agcagcgctc ggtatcgatg cggggaggcg gcccaaaggg
      781 agatccgact cgtctgaggg cgaaggcgaa gacgcggaag aggccgcaga gccggcagca
      841 ggccgcggga aggaaggtcc gctggattga gggccgaagg gacgtagcag aaggacgtcc
      901 cgcgcagaat ccaggtggca acataggcga gcagccaagg aaaggacgat gatttccccg
      961 acaacaccac ggaattgtca gtgcccaaca gccgagcccc tgtccagcag cgggcaaggc
     1021 aggcggcgat gagttccgcc gtggcaatag ggagggggaa agcgaaagtc ccggaaagga
     1081 gctgacaggt ggtggcaatg ccccaaccag tgggggttgc gtcagcaaac acagtgcaca
     1141 ccacgccacg ttgcctgaca acgggccaca actcctcata aagagacagc aaccaggatt
     1201 tatacaagga ggagaaaatg aaagccatac gggaagcaat agcatgatac aaaggcatta
     1261 aagcagcgta tccacatagc gtaaaaggag caacatagtt aagaatacca gtcaatcttt
     1321 cacaaatttt gtaatccaga ggttgattat cgataagctt gatatcttac tctctggcct
     1381 cttgggagat ctctttcagg ttgttgacct cgttggccag ctgccggtta ggccgcaggt
     1441 tcttcagcag gaatctctgt ctgcacacag ggcacacaga tccgccgcct ttgccgactt
     1501 ggctgatgca ctcttggcag aagctgtggc cgcattcgat ggacacgggt tccacgaagg
     1561 ggtccagaca cctaggacag gtcacttctt cccacatcat ggtcagtctg gcggcagagg
     1621 ccatcgatcc gccaccaccg gagccgaatt ccaaaccctg cttggccagg gaggcagaca
     1681 cctcgtcagc tagcgtggcg agctggggcg agtctaccat gtcgatgctg ccggtggagg
     1741 agacattgct gagatgccgt ggagacgtgt ccccagacac cactggcgac ttgtacacga
     1801 tctccgcccc gtggtctgtc ttggctttgg cgttctcgcg gaaggtcagc ttgtgggttt
     1861 caatcttttt atttcctccg ccagggacgt gggtgatatt gtccagggac ccaatcttcg
     1921 actggactct gtccttgaag tcaagcttct cagattttac ttccacctgg ccacctcctg
     1981 gtttatgatg gatgttgcct aatgagccac acttagaggt caccttgctc aggtcaactg
     2041 gtttgtagac tatttgcaca ctgccgcctc cactgacgtg tttgatatta tcctttgatc
     2101 cacacttgga ctggacgttg ctaagatcca gcttcttatt aattatctgc accttcccgc
     2161 ctcccggctg gtgcttcagg ttctcagtgg agccgatctt ggacttgaca ttcttcaggt
     2221 ctggcatggg cacgggggct gtctgcaggc ggctcttggc ggaagacggc gacttgggtg
     2281 gagtacggac cactgccacc ttcttgggct cccgggtggg tggggttgga agggacgggg
     2341 tacgggagcg gctgccggga gtgcctgggg agccggggct gctgtagccg ctgcgatccc
     2401 ctgattttgg aggttcacca gagctgggtg gtgtctttgg agcgggcggg gtttttgctg
     2461 gaatcctggt ggcgttggcc tggcccttct ggcctggagg ggctgctccc cgcggtgtgg
     2521 cgatcttcgt tttaccatca gcccccttgg cttttttgtc atcgcttcca gtcccgtctt
     2581 tgcttttact gaccatgcga gcttgggtca cgtgaccagc agcttcgtct tccaggctgg
     2641 gggtgtctcc aatgcctgct tcttcagctt tcaggccagc gtccgtgtca ccctcttggt
     2701 cttggtgcat ggtgtagccc ccctgatctt tcctgtcccc caacccgtac gtcccagcgt
     2761 gatcttccat cacttcgaac tcctggcggg gctcagccat cttaattaaa ggtccagggt
     2821 tctcctccac gtctccagcc tgcttcagca ggctgaagtt agtagctccg cttccgaatt
     2881 ccttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga
     2941 ccatgtgatc gcgcttctcg ttggggtctt tgctcagggc ggactggtag ctcaggtagt
     3001 ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg
     3061 cgagctgcac gccgccgtcc tcgatgttgt ggcggatctt gaagttggcc ttgatgccgt
     3121 tcttctgctt gtcggcggtg atatagacgt tgtggctgtt gtagttgtac tccagcttgt
     3181 gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca
     3241 gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga
     3301 agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct
     3361 tcatgtggtc ggggtagcgg gcgaagcact gcaggccgta gcccagggtg gtcacgaggg
     3421 tgggccaggg cacgggcagc ttgccggtgg tgcagatcag cttcagggtc agcttgccgt
     3481 aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt
     3541 ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgg
     3601 taccttctcg actgcgctct caggcacgac acgactcctc cgctgcccac cgcagactga
     3661 ggcagcgctg agtcgccggc gccgcagcgc agatggtcgc gcccgtgccc ccctatctcg
     3721 cgcctcgcgt ggtgcggtcc ggctgggccg gcggcggcgc ggacgcgacc aaggtggccg
     3781 ggaaggggag tttgcggggg accggcgagt gacgtcagcg cgccttcagt gctgaggcgg
     3841 cggtggcgcg cgccgccagg cgggggcgaa ggcactgtcc gcggtgctga agctggcagt
     3901 gcgcacgcgc ctcgccgcat cctgtttccc ctccccctct ctgatagggg atgcgcaatt
     3961 tggggaatgg gggttgggtg cttgtccagt gggtcggggt cggtcgtcag gtaggcaccc
     4021 ccaccccgcc tcatcctggt cctaaaaccc acttgcactc atacgcaggg ccctctgcag
     4081 tctagacaca cgcgtgcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
     4141 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggcggcct
     4201 cagtgagcga gcgagcgcgc agctgcctgc aggacatgtg agcaaaaggc cagcaaaagg
     4261 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4321 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4381 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4441 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4501 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4561 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4621 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4681 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     4741 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4801 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4861 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4921 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4981 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5041 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5101 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     5161 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5221 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5281 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5341 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5401 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5461 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5521 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5581 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5641 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5701 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5761 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5821 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5881 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5941 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6001 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     6061 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     6121 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     6181 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     6241 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     6301 aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     6361 tttttaacca atagaccgaa atcggcaaaa tcccttataa atcaaaagaa tagcccgaga
     6421 tagagttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     6481 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccca
     6541 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     6601 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     6661 cgaaaggagc gggcgctaag gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     6721 cacccgccgc gcttaatgcg ccgctacagg gcgcgtact
//
