LOCUS       sequence-426883-        8567 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV.CBA.eGFP.2A.wtTau propagation reporter WT-2N4R-hTau.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8567)
  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 8567)
  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.wtTau
FEATURES             Location/Qualifiers
     source          1..8567
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(170..189)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        256..635
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        637..915
                     /label=chicken beta-actin promoter
     primer_bind     1328..1350
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1412..1431
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     misc_feature    1457..1509
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      1517..1526
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1523..1525,1526..1528,1529..2239)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1523..1525,1526..1528,1529..2239)
                     /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(1568..1589)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1829..1848)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2176..2197
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2249..2302
                     /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    3653..4241
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3706..3726)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4124..4135)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4333..4352)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4398..4453
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4452..4471)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     repeat_region   4853..4982
                     /label=AAV2 ITR
     rep_origin      5353..5866
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5440..5459)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5650..5671
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6348..6367)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6467..6489
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6527..6545)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6613..6717
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6718..6786,6787..7578)
                     /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(6936..6955)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7749..8337
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8238..8257
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgcagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcggcctcag
       61 tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt tccttgtagt
      121 taatgattaa cccgccatgc tacttatcta cgtagccatg ctctagcggc ctcggcctct
      181 gcataaataa aaaaaattag tcagccatga gcttggccca ttgcatacgt tgtatccata
      241 tcataatatg tactcgacat tgattattga ctagttatta atagtaatca attacggggt
      301 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
      361 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
      421 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
      481 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
      541 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
      601 agtacatcta cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct
      661 tcactctccc catctccccc ccctccccac ccccaatttt gtatttattt attttttaat
      721 tattttgtgc agcgatgggg gcgggggggg gggggggggc gcgcgccagg cggggcgggg
      781 cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc
      841 gcgctccgaa agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga
      901 agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc
      961 gcctcgcgcc gcccgccccg gctctgactg accgcgttac tcccacaggt gagcgggcgg
     1021 gacggccctt ctcctccggg ctgtaattag cgcttggttt aatgacggct tgtttctttt
     1081 ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc
     1141 tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc
     1201 ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag
     1261 gggagcgcgg ccgggggcgg tgccccgcgg ggggacggct gccttcgggg gggacggggc
     1321 agggcggggt tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt
     1381 catgccttct tctttttcct acagctcctg ggcaacgtgc tggttattgt gctgtctcat
     1441 cattttggca aagaatctcg agctcaagct tcgaattctg cagtcgacgg taccgcgggc
     1501 ccgggatcca ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
     1561 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     1621 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     1681 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     1741 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     1801 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     1861 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     1921 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     1981 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     2041 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     2101 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     2161 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     2221 catggacgag ctgtacaagg gcagtggaga gggcagagga agtctgctaa catgcggtga
     2281 cgtcgaggag aatcctggcc cagctgagcc ccgccaggag ttcgaagtga tggaagatca
     2341 cgctgggacg tacgggttgg gggacaggaa agatcagggg ggctacacca tgcaccaaga
     2401 ccaagagggt gacacggacg ctggcctgaa agaatctccc ctgcagaccc ccactgagga
     2461 cggatctgag gaaccgggct ctgaaacctc tgatgctaag agcactccaa cagcggaaga
     2521 tgtgacagca cccttagtgg atgagggagc tcccggcaag caggctgccg cgcagcccca
     2581 cacggagatc ccagaaggaa ccacagctga agaagcaggc attggagaca cccccagcct
     2641 ggaagacgaa gctgctggtc acgtgaccca agctcgcatg gtcagtaaaa gcaaagacgg
     2701 gactggaagc gatgacaaaa aagccaaggg ggctgatggt aaaacgaaga tcgccacacc
     2761 gcggggagca gcccctccag gccagaaggg ccaggccaac gccaccagga ttccagcaaa
     2821 aaccccgccc gctccaaaga caccacccag ctctggtgaa cctccaaaat caggggatcg
     2881 cagcggctac agcagccccg gctccccagg cactcccggc agccgctccc gcaccccgtc
     2941 ccttccaacc ccacccaccc gggagcccaa gaaggtggca gtggtccgta ctccacccaa
     3001 gtcgccgtct tccgccaaga gccgcctgca gacagccccc gtgcccatgc cagacctgaa
     3061 gaatgtcaag tccaagatcg gctccactga gaacctgaag caccagccgg gaggcgggaa
     3121 ggtgcagata attaataaga agctggatct tagcaacgtc cagtccaagt gtggctcaaa
     3181 ggataatatc aaacacgtcc cgggaggcgg cagtgtgcaa atagtctaca aaccagttga
     3241 cctgagcaag gtgacctcca agtgtggctc attaggcaac atccatcata aaccaggagg
     3301 tggccaggtg gaagtaaaat ctgagaagct tgacttcaag gacagagtcc agtcgaagat
     3361 tgggtccctg gacaatatca cccacgtccc tggcggagga aataaaaaga ttgaaaccca
     3421 caagctgacc ttccgcgaga acgccaaagc caagacagac cacggggcgg agatcgtgta
     3481 caagtcgcca gtggtgtctg gggacacgtc tccacggcat ctcagcaatg tctcctccac
     3541 cggcagcatc gacatggtag actcgcccca gctcgccacg ctagctgacg aggtgtctgc
     3601 ctccctggcc aagcagggtt tgtagactag cgatatcggt accggatccg ataatcaacc
     3661 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     3721 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     3781 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     3841 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     3901 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     3961 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     4021 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     4081 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     4141 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     4201 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcggatcc aattcactcc
     4261 tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa
     4321 ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga
     4381 gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt
     4441 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga
     4501 gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg
     4561 gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag
     4621 aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt
     4681 aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact
     4741 actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgacctgc agcccaagct
     4801 ctagagcatg gctacgtaga taagtagcat ggcgggttaa tcattaacta caaggaaccc
     4861 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     4921 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     4981 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     5041 aatggcgaat ggcgattccg ttgcaatggc tggcggtaat attgttctgg atattaccag
     5101 caaggccgat agtttgagtt cttctactca ggcaagtgat gttattacta atcaaagaag
     5161 tattgcgaca acggttaatt tgcgtgatgg acagactctt ttactcggtg gcctcactga
     5221 ttataaaaac acttctcagg attctggcgt accgttcctg tctaaaatcc ctttaatcgg
     5281 cctcctgttt agctcccgct ctgattctaa cgaggaaagc acgttatacg tgctcgtcaa
     5341 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     5401 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     5461 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     5521 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     5581 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     5641 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     5701 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     5761 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttaa atatttgctt
     5821 atacaatctt cctgtttttg gggcttttct gattatcaac cggggtacat atgattgaca
     5881 tgctagtttt acgattaccg ttcatcgatt ctcttgtttg ctccagactc tcaggcaatg
     5941 acctgatagc ctttgtagag acctctcaaa aatagctacc ctctccggca tgaatttatc
     6001 agctagaacg gttgaatatc atattgatgg tgatttgact gtctccggcc tttctcaccc
     6061 gtttgaatct ttacctacac attactcagg cattgcattt aaaatatatg agggttctaa
     6121 aaatttttat ccttgcgttg aaataaaggc ttctcccgca aaagtattac agggtcataa
     6181 tgtttttggt acaaccgatt tagctttatg ctctgaggct ttattgctta attttgctaa
     6241 ttctttgcct tgcctgtatg atttattgga tgttggaatc gcctgatgcg gtattttctc
     6301 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     6361 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     6421 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     6481 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     6541 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     6601 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     6661 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     6721 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     6781 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     6841 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     6901 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     6961 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     7021 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     7081 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     7141 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     7201 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     7261 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     7321 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     7381 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     7441 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     7501 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     7561 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     7621 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     7681 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     7741 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     7801 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     7861 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     7921 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     7981 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     8041 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     8101 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     8161 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     8221 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     8281 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     8341 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     8401 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     8461 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     8521 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcatta
//
