LOCUS       sequence-426887-        8568 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV.CBA.eGFP.2A.P301L-Tau propagation reporter P301L-2N4R-hTau.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8568)
  AUTHORS   Wegmann S, Bennett RE, Delorme L, Robbins AB, Hu M, McKenzie D, Kirk
            MJ, Schiantarelli J, Tunio N, Amaral AC, Fan Z, Nicholls S, Hudry E,
            Hyman BT
  TITLE     Experimental evidence for the age dependence of tau protein spread 
            in the brain.
  JOURNAL   Sci Adv. 2019 Jun 26;5(6):eaaw6404. doi: 10.1126/sciadv.aaw6404. 
            eCollection 2019 Jun.
  PUBMED    31249873
REFERENCE   2  (bases 1 to 8568)
  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: "Sci Adv."; 
            date: "2019-06-26"; volume: "5(6):eaaw6404. doi"; pages: " 
            10.1126/sciadv.aaw6404. eCollection 2019 Jun"
COMMENT     Sequence Label: AAV.CBA.eGFP.2A.P301L-Tau
FEATURES             Location/Qualifiers
     source          1..8568
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(102..121)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        188..567
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        569..848
                     /label=chicken beta-actin promoter
     primer_bind     1261..1283
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1345..1364
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     misc_feature    1390..1442
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      1450..1459
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1456..1458,1459..1461,1462..2172)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1456..1458,1459..1461,1462..2172)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1501..1522)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1762..1781)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2109..2130
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2182..2235
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     misc_feature    3586..4174
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3639..3659)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4057..4068)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4266..4285)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4331..4386
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4385..4404)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     repeat_region   4786..4915
                     /label=AAV2 ITR
     rep_origin      5286..5799
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5373..5392)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5583..5604
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6281..6300)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6400..6422
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6460..6478)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6546..6650
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6651..6719,6720..7511)
                     /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(6869..6888)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7682..8270
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8171..8190
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgagcgcgca gagagggagt ggccaactcc atcactaggg gttccttgta gttaatgatt
       61 aacccgccat gctacttatc tacgtagcca tgctctagcg gcctcggcct ctgcataaat
      121 aaaaaaaatt agtcagccat gagcttggcc cattgcatac gttgtatcca tatcataata
      181 tgtactcgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
      241 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      301 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      361 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      421 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      481 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      541 tacgtattag tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc
      601 cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt
      661 gcagcgatgg gggcgggggg gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg
      721 aggggcgggg cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc
      781 gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc
      841 ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc
      901 gccgcccgcc ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc
      961 cttctcctcc gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc
     1021 tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggg
     1081 gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct
     1141 gtgagcgctg cgggcgcggc gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg
     1201 cggccggggg cggtgccccg cggggggacg gctgccttcg ggggggacgg ggcagggcgg
     1261 ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat gttcatgcct
     1321 tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg
     1381 gcaaagaatc tcgagctcaa gcttcgaatt ctgcagtcga cggtaccgcg ggcccgggat
     1441 ccaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     1501 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     1561 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     1621 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     1681 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1741 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1801 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1861 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     1921 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     1981 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     2041 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     2101 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     2161 gagctgtaca agggcagtgg agagggcaga ggaagtctgc taacatgcgg tgacgtcgag
     2221 gagaatcctg gcccagctga gccccgccag gagttcgaag tgatggaaga tcacgctggg
     2281 acgtacgggt tgggggacag gaaagatcag gggggctaca ccatgcacca agaccaagag
     2341 ggtgacacgg acgctggcct gaaagaatct cccctgcaga cccccactga ggacggatct
     2401 gaggaaccgg gctctgaaac ctctgatgct aagagcactc caacagcgga agatgtgaca
     2461 gcacccttag tggatgaggg agctcccggc aagcaggctg ccgcgcagcc ccacacggag
     2521 atcccagaag gaaccacagc tgaagaagca ggcattggag acacccccag cctggaagac
     2581 gaagctgctg gtcacgtgac ccaagctcgc atggtcagta aaagcaaaga cgggactgga
     2641 agcgatgaca aaaaagccaa gggggctgat ggtaaaacga agatcgccac accgcgggga
     2701 gcagcccctc caggccagaa gggccaggcc aacgccacca ggattccagc aaaaaccccg
     2761 cccgctccaa agacaccacc cagctctggt gaacctccaa aatcagggga tcgcagcggc
     2821 tacagcagcc ccggctcccc aggcactccc ggcagccgct cccgcacccc gtcccttcca
     2881 accccaccca cccgggagcc caagaaggtg gcagtggtcc gtactccacc caagtcgccg
     2941 tcttccgcca agagccgcct gcagacagcc cccgtgccca tgccagacct gaagaatgtc
     3001 aagtccaaga tcggctccac tgagaacctg aagcaccagc cgggaggcgg gaaggtgcag
     3061 ataattaata agaagctgga tcttagcaac gtccagtcca agtgtggctc aaaggataat
     3121 atcaaacacg tcctgggagg cggcagtgtg caaatagtct acaaaccagt tgacctgagc
     3181 aaggtgacct ccaagtgtgg ctcattaggc aacatccatc ataaaccagg aggtggccag
     3241 gtggaagtaa aatctgagaa gcttgacttc aaggacagag tccagtcgaa gattgggtcc
     3301 ctggacaata tcacccacgt ccctggcgga ggaaataaaa agattgaaac ccacaagctg
     3361 accttccgcg agaacgccaa agccaagaca gaccacgggg cggagatcgt gtacaagtcg
     3421 ccagtggtgt ctggggacac gtctccacgg catctcagca atgtctcctc caccggcagc
     3481 atcgacatgg tagactcgcc ccagctcgcc acgctagctg acgaggtgtc tgcctccctg
     3541 gccaagcagg gtttgtagac tagcgatatc ggtaccggat ccgataatca acctctggat
     3601 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     3661 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     3721 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     3781 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     3841 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     3901 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     3961 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
     4021 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     4081 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     4141 acgagtcgga tctccctttg ggccgcctcc ccgcatcgga tccaattcac tcctcaggtg
     4201 caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac
     4261 tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg
     4321 acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt
     4381 ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg
     4441 gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa
     4501 agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc
     4561 ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc
     4621 ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca
     4681 gtcatagctg tccctcttct cttatggaga tccctcgacc tgcagcccaa gctctagagc
     4741 atggctacgt agataagtag catggcgggt taatcattaa ctacaaggaa cccctagtga
     4801 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     4861 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctggc
     4921 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     4981 aatggcgatt ccgttgcaat ggctggcggt aatattgttc tggatattac cagcaaggcc
     5041 gatagtttga gttcttctac tcaggcaagt gatgttatta ctaatcaaag aagtattgcg
     5101 acaacggtta atttgcgtga tggacagact cttttactcg gtggcctcac tgattataaa
     5161 aacacttctc aggattctgg cgtaccgttc ctgtctaaaa tccctttaat cggcctcctg
     5221 tttagctccc gctctgattc taacgaggaa agcacgttat acgtgctcgt caaagcaacc
     5281 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     5341 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     5401 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     5461 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     5521 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     5581 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     5641 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     5701 atttaacgcg aattttaaca aaatattaac gcttacaatt taaatatttg cttatacaat
     5761 cttcctgttt ttggggcttt tctgattatc aaccggggta catatgattg acatgctagt
     5821 tttacgatta ccgttcatcg attctcttgt ttgctccaga ctctcaggca atgacctgat
     5881 agcctttgta gagacctctc aaaaatagct accctctccg gcatgaattt atcagctaga
     5941 acggttgaat atcatattga tggtgatttg actgtctccg gcctttctca cccgtttgaa
     6001 tctttaccta cacattactc aggcattgca tttaaaatat atgagggttc taaaaatttt
     6061 tatccttgcg ttgaaataaa ggcttctccc gcaaaagtat tacagggtca taatgttttt
     6121 ggtacaaccg atttagcttt atgctctgag gctttattgc ttaattttgc taattctttg
     6181 ccttgcctgt atgatttatt ggatgttgga atcgcctgat gcggtatttt ctccttacgc
     6241 atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg
     6301 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     6361 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     6421 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     6481 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     6541 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     6601 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     6661 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     6721 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     6781 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     6841 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     6901 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     6961 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     7021 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     7081 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     7141 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     7201 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     7261 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     7321 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     7381 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     7441 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     7501 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     7561 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     7621 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     7681 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     7741 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     7801 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     7861 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     7921 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     7981 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     8041 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     8101 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     8161 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     8221 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     8281 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     8341 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     8401 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     8461 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgc gcgctcgctc
     8521 gctcactgag gccgcccggg caaagcccgg gcggcctcag tgagcgag
//
