LOCUS       sequence-466808-        6759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of Venus-P2A-Tau-RING 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
FEATURES             Location/Qualifiers
     source          1..6759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     22..41
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(260..364)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     432..450
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(488..510)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     610..629
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(646..1101)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(783..804)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     995..1014
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1176..1316
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1187..1316
                     /label=AAV2 ITR
     polyA_signal    1356..1832
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     1614..1633
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    1864..2452
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             1970..1981
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2379..2399
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3914..3970)
                     /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(3986..4696,4697..4699,4700..4702))
                     /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(4028..4049)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4377..4396
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4636..4657
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(4716..5163)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   5208..5337
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(5399..5987)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5479..5498)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6158..6759,1..190,191..259))
                     /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"
ORIGIN
        1 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
       61 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
      121 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
      181 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
      241 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
      301 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
      361 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
      421 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
      481 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
      541 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
      601 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccataaaatt gtaaacgtta
      661 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaataga
      721 ccgaaatcgg caaaatccct tataaatcaa aagaatagcc cgagatagag ttgagtgttg
      781 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
      841 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc acccaaatca agttttttgg
      901 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
      961 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     1021 ctaaggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     1081 atgcgccgct acagggcgcg tactatggtt gctttgacgt atgcggtgtg aaataccgca
     1141 cagatgcgta aggagaaaat accgcatcag gcgcccctgc aggcagctgc gcgctcgctc
     1201 gctcactgag gccgcccggg caaagcccgg gcgtcgggcg acctttggtc gcccggcctc
     1261 agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg
     1321 ccgctcggtc cgcacgtggt tacctacaaa atcagaagga cagggaaggg agcagtggtt
     1381 cacgcctgta atcccagcaa tttgggaggc caaggtgggt agatcacctg agattaggag
     1441 ttggagacca gcctggccaa tatggtgaaa ccccgtctct accaaaaaaa caaaaattag
     1501 ctgagcctgg tcatgcatgc ctggaatccc aacaactcgg gaggctgagg caggagaatc
     1561 gcttgaaccc aggaggcgga gattgcagtg agccaagatt gtgccactgc actccagctt
     1621 ggttcccaat agaccccgca ggccctacag gttgtcttcc caacttgccc cttgctccat
     1681 accacccccc tccaccccat aatattatag aaggacacct agtcagacaa aatgatgcaa
     1741 cttaatttta ttaggacaag gctggtgggc actggagtgg caacttccag ggccaggaga
     1801 ggcactgggg aggggtcaca gggatgccac ccgtagatct ctcgagcagc gctcggtatc
     1861 gatgcgggga ggcggcccaa agggagatcc gactcgtctg agggcgaagg cgaagacgcg
     1921 gaagaggccg cagagccggc agcaggccgc gggaaggaag gtccgctgga ttgagggccg
     1981 aagggacgta gcagaaggac gtcccgcgca gaatccaggt ggcaacatag gcgagcagcc
     2041 aaggaaagga cgatgatttc cccgacaaca ccacggaatt gtcagtgccc aacagccgag
     2101 cccctgtcca gcagcgggca aggcaggcgg cgatgagttc cgccgtggca atagggaggg
     2161 ggaaagcgaa agtcccggaa aggagctgac aggtggtggc aatgccccaa ccagtggggg
     2221 ttgcgtcagc aaacacagtg cacaccacgc cacgttgcct gacaacgggc cacaactcct
     2281 cataaagaga cagcaaccag gatttataca aggaggagaa aatgaaagcc atacgggaag
     2341 caatagcatg atacaaaggc attaaagcag cgtatccaca tagcgtaaaa ggagcaacat
     2401 agttaagaat accagtcaat ctttcacaaa ttttgtaatc cagaggttga ttatcgataa
     2461 gcttgatatc ttactctctg gcctcctggc tgatttcttt aaggttgttc accatgttgg
     2521 ctagctgtcg attgggccgg agattcttga gcagaaagcg ctgccggcac acaggacaga
     2581 cgctgccccc acctttccca acctgagaga tgcattcctg gcagaagctg tggccacact
     2641 cgatgctcac aggctccacg aaggggtcca ggcagatagg gcatgtgacc tcctcccaca
     2701 tcattgtcaa gcgtgctgct gaagccatgc ctgagccacc tccggatccg aattccaaac
     2761 cctgcttggc cagggaggca gacacctcgt cagctagcgt ggcgagctgg ggcgagtcta
     2821 ccatgtcgat gctgccggtg gaggagacat tgctgagatg ccgtggagac gtgtccccag
     2881 acaccactgg cgacttgtac acgatctccg ccccgtggtc tgtcttggct ttggcgttct
     2941 cgcggaaggt cagcttgtgg gtttcaatct ttttatttcc tccgccaggg acgtgggtga
     3001 tattgtccag ggacccaatc ttcgactgga ctctgtcctt gaagtcaagc ttctcagatt
     3061 ttacttccac ctggccacct cctggtttat gatggatgtt gcctaatgag ccacacttgg
     3121 aggtcacctt gctcaggtca actggtttgt agactatttg cacactgccg cctccactga
     3181 cgtgtttgat attatccttt gagccacact tggactggac gttgctaaga tccagcttct
     3241 tattaattat ctgcaccttc ccgcctcccg gctggtgctt caggttctca gtggagccga
     3301 tcttggactt gacattcttc aggtctggca tgggcacggg ggctgtctgc aggcggctct
     3361 tggcggaaga cggcgacttg ggtggagtac ggaccactgc caccttcttg ggctcccggg
     3421 tgggtggggt tggaagggac ggggtgcggg agcggctgcc gggagtgcct ggggagccgg
     3481 ggctgctgta gccgctgcga tcccctgatt ttggaggttc accagagctg ggtggtgtct
     3541 ttggagcggg cggggttttt gctggaatcc tggtggcgtt ggcctggccc ttctggcctg
     3601 gaggggctgc tccccgcggt gtggcgatct tcgttttacc atcagccccc ttggcttttt
     3661 tgtcatcgct tccagtcccg tctttgcttt tactgaccat gcgagcttgg gtcacgtgac
     3721 cagcagcttc gtcttccagg ctgggggtgt ctccaatgcc tgcttcttca gctttcaggc
     3781 cagcgtccgt gtcaccctct tggtcttggt gcatggtgta gcccccctga tctttcctgt
     3841 cccccaaccc gtacgtccca gcgtgatctt ccatcacttc gaactcctgg cggggctcag
     3901 ccatcttaat taaaggtcca gggttctcct ccacgtctcc agcctgcttc agcaggctga
     3961 agttagtagc tccgcttccg aattccttgt acagctcgtc catgccgaga gtgatcccgg
     4021 cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg tctttgctca
     4081 gggcggactg gtagctcagg tagtggttgt cgggcagcag cacggggccg tcgccgatgg
     4141 gggtgttctg ctggtagtgg tcggcgagct gcacgccgcc gtcctcgatg ttgtggcgga
     4201 tcttgaagtt ggccttgatg ccgttcttct gcttgtcggc ggtgatatag acgttgtggc
     4261 tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc
     4321 ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg gcgcgggtct
     4381 tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg
     4441 cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcgggcgaag cactgcaggc
     4501 cgtagcccag ggtggtcacg agggtgggcc agggcacggg cagcttgccg gtggtgcaga
     4561 tcagcttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg gacacgctga
     4621 acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc ccggtgaaca
     4681 gctcctcgcc cttgctcacc atggtacctt ctcgactgcg ctctcaggca cgacacgact
     4741 cctccgctgc ccaccgcaga ctgaggcagc gctgagtcgc cggcgccgca gcgcagatgg
     4801 tcgcgcccgt gcccccctat ctcgcgcctc gcgtggtgcg gtccggctgg gccggcggcg
     4861 gcgcggacgc gaccaaggtg gccgggaagg ggagtttgcg ggggaccggc gagtgacgtc
     4921 agcgcgcctt cagtgctgag gcggcggtgg cgcgcgccgc caggcggggg cgaaggcact
     4981 gtccgcggtg ctgaagctgg cagtgcgcac gcgcctcgcc gcatcctgtt tcccctcccc
     5041 ctctctgata ggggatgcgc aatttgggga atgggggttg ggtgcttgtc cagtgggtcg
     5101 gggtcggtcg tcaggtaggc acccccaccc cgcctcatcc tggtcctaaa acccacttgc
     5161 actcatacgc agggccctct gcagtctaga cacacgcgtg cggccgcagg aacccctagt
     5221 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     5281 ggtcgcccga cgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggaca
     5341 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5401 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5461 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5521 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5581 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5641 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5701 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5761 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5821 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5881 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5941 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6001 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6061 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6121 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6181 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6241 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6301 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6361 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6421 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6481 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6541 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6601 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6661 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6721 agtcattctg agaatagtgt atgcggcgac cgagttgct
//
