LOCUS       sequence-346595-        6917 bp    DNA     circular SYN 14-APR-2025
DEFINITION  This AAV plasmid vector expresses human 2N4R microtubule-associated 
            protein tau with P301L mutation..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6917)
  AUTHORS   You Y, Botros MB, Enoo AAV, Bockmiller A, Herron S, Delpech JC, 
            Ikezu T
  TITLE     Cre-inducible Adeno Associated Virus-mediated Expression of P301L 
            Mutant Tau Causes Motor Deficits and Neuronal Degeneration in the 
            Substantia Nigra.
  JOURNAL   Neuroscience. 2019 Dec 1;422:65-74. doi: 
            10.1016/j.neuroscience.2019.10.001. Epub 2019 Nov 2.
  PUBMED    31689387
REFERENCE   2  (bases 1 to 6917)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Neuroscience."; date: "2019-12-1"; volume: "422:65-74. doi"; pages:
            " 10.1016/j.neuroscience.2019.10.001. Epub 2019 Nov 2"
COMMENT     Sequence Label: pAAV-FLEX-P301L Tau
FEATURES             Location/Qualifiers
     source          1..6917
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   26..155
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        256..1434
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          487..1425
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1382..1402
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1465..1498)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(1549..1582)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      2913..2922
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2927..2960
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    3011..3044
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3069..3657
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3122..3142)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3540..3551)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3689..4165
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3888..3907)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4205..4345
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4205..4334
                     /label=AAV2 ITR
     rep_origin      4420..4875
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4507..4526)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4717..4738
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4892..4911)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5011..5033
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5071..5089)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5157..5261
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5262..5330,5331..6122)
                     /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(5480..5499)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6293..6881
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6782..6801
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttttgctggc cttttgctca catgtcctgc aggcagctgc gcgctcgctc gctcactgag
       61 gccgcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag
      121 agagggagtg gccaactcca tcactagggg ttcctgcggc cgcacgcgta agctttgcaa
      181 agatggataa agttttaaac agagaggaat ctttgcagct aatggacctt ctaggtcttg
      241 aaaggagtgg gaattggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc
      301 cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg
      361 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa
      421 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga
      481 acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc tctttacggg ttatggccct
      541 tgcgtgcctt gaattacttc cacctggctg cagtacgtga ttcttgatcc cgagcttcgg
      601 gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct
      661 tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc
      721 gcctgtctcg ctgctttcga taagtctcta gccatttaaa atttttgatg acctgctgcg
      781 acgctttttt tctggcaaga tagtcttgta aatgcgggcc aagatctgca cactggtatt
      841 tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg
      901 aggcggggcc tgcgagcgcg gccaccgaga atcggacggg ggtagtctca agctggccgg
      961 cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg
     1021 gcccggtcgg caccagttgc gtgagcggaa agatggccgc ttcccggccc tgctgcaggg
     1081 agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg
     1141 aaaagggcct ttccgtcctc agccgtcgct tcatgtgact ccacggagta ccgggcgccg
     1201 tccaggcacc tcgattagtt ctcgagcttt tggagtacgt cgtctttagg ttggggggag
     1261 gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt taggccagct
     1321 tggcacttga tgtaattctc cttggaattt gccctttttg agtttggatc ttggttcatt
     1381 ctcaagcctc agacagtggt tcaaagtttt tttcttccat ttcaggtgtc gtgaggtacc
     1441 ggatcctcta gagtcgactc cggaataact tcgtatagga tactttatac gaagttatgc
     1501 agaatggtag ctggattgta gctgctatta gcaatatgaa acctcttaat aacttcgtat
     1561 agcatacatt atacgaagtt atggcgcgcc tcacaaaccc tgcttggcca gggaggcaga
     1621 cacctcgtca gctagcgtgg cgagctgggg cgagtctacc atgtcgatgc tgccggtgga
     1681 ggagacattg ctgagatgcc gtggagacgt gtccccagac accactggcg acttgtacac
     1741 gatctccgcc ccgtggtctg tcttggcttt ggcgttctcg cggaaggtca gcttgtgggt
     1801 ttcaatcttt ttatttcctc cgccagggac gtgggtgata ttgtccaggg acccaatctt
     1861 cgactggact ctgtccttga agtcaagctt ctcagatttt acttccacct ggccacctcc
     1921 tggtttatga tggatgttgc ctaatgagcc acacttggag gtcaccttgc tcaggtcaac
     1981 tggtttgtag actatttgca cactgccgcc tcccaggacg tgtttgatat tatcctttga
     2041 gccacacttg gactggacgt tgctaagatc cagcttctta ttaattatct gcaccttccc
     2101 gcctcccggc tggtgcttca ggttctcagt ggagccgatc ttggacttga cattcttcag
     2161 gtctggcatg ggcacggggg ctgtctgcag gcggctcttg gcggaagacg gcgacttggg
     2221 tggagtacgg accactgcca ccttcttggg ctcccgggtg ggtggggttg gaagggacgg
     2281 ggtgcgggag cggctgccgg gagtgcctgg ggagccgggg ctgctgtagc cgctgcgatc
     2341 ccctgatttt ggaggttcac cagagctggg tggtgtcttt ggagcgggcg gggtttttgc
     2401 tggaatcctg gtggcgttgg cctggccctt ctggcctgga ggggctgctc cccgcggtgt
     2461 ggcgatcttc gttttaccat cagccccctt ggcttttttg tcatcgcttc cagtcccgtc
     2521 tttgctttta ctgaccatgc gagcttgggt cacgtgacca gcagcttcgt cttccaggct
     2581 gggggtgtct ccaatgcctg cttcttcagc tgtggttcct tctgggatct ccgtgtgggg
     2641 ctgcgcggca gcctgcttgc cgggagctcc ctcatccact aagggtgctg tcacatcttc
     2701 cgctgttgga gtgctcttag catcagaggt ttcagagccc ggttcctcag atccgtcctc
     2761 agtgggggtc tgcaggggag attctttcag gccagcgtcc gtgtcaccct cttggtcttg
     2821 gtgcatggtg tagcccccct gatctttcct gtcccccaac ccgtacgtcc cagcgtgatc
     2881 ttccatcact tcgaactcct ggcggggctc agccatggtg gctagcataa cttcgtataa
     2941 agtatcctat acgaagttat ttgccttaac ccagaaatta tcactgttat tctttagaat
     3001 ggtgcaaaga ataacttcgt ataatgtatg ctatacgaag ttatgaattc gatatcaagc
     3061 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     3121 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     3181 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     3241 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     3301 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     3361 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     3421 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     3481 ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     3541 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     3601 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc
     3661 gataccgagc gctgctcgag agatctacgg gtggcatccc tgtgacccct ccccagtgcc
     3721 tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag
     3781 ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt
     3841 atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc
     3901 aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc
     3961 gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag
     4021 ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa
     4081 ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg
     4141 aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg gaccgagcgg
     4201 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     4261 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     4321 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     4381 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     4441 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag
     4501 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4561 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4621 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4681 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4741 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     4801 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4861 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     4921 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     4981 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     5041 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     5101 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     5161 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     5221 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     5281 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5341 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     5401 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     5461 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5521 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5581 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5641 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5701 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5761 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5821 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5881 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5941 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     6001 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     6061 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     6121 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     6181 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6241 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6301 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6361 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6421 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6481 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6541 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6601 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6661 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6721 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6781 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6841 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6901 ttttacggtt cctggcc
//
