LOCUS       Exported                8418 bp ds-DNA     circular SYN 27-MAR-2024
DEFINITION  AAV.CBA.eGFP.2A.P301L-Tau propagation reporter P301L-2N4R-hTau.
ACCESSION   .
VERSION     .
KEYWORDS    AAV.CBA.eGFP.2A.P301L-Tau
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8418)
  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 8418)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 27, 2024 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8418
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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..825
                     /label=chicken beta-actin promoter
                     /note="contains a 21-bp deletion relative to the standard 
                     chicken beta-actin promoter"
     primer_bind     1238..1260
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1322..1341
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     misc_feature    1367..1419
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      1427..1436
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1433..2149
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             1433..2149
                     /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(1478..1499)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1739..1758)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2086..2107
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2159..2212
                     /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    3563..4151
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3616..3636)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4034..4045)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4243..4262)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4308..4363
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4362..4381)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     repeat_region   4763..4892
                     /label=AAV2 ITR
     rep_origin      5263..5776
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5350..5369)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5560..5581
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6258..6277)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6377..6399
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6437..6455)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6523..6627
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6628..7488
                     /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(6846..6865)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7659..8247
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8148..8167
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8401..8418
                     /label=L4440
                     /note="L4440 vector, 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 sggggcgrgg ggsggggsgg ggsgrggcgg
      721 agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg
      781 cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc
      841 tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg
      901 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag
      961 cgcttggttt aatgacggct tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc
     1021 cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg
     1081 gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg
     1141 gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg
     1201 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1261 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1321 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaatctcg agctcaagct
     1381 tcgaattctg cagtcgacgg taccgcgggc ccgggatcca ccggtcgcca ccatggtgag
     1441 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     1501 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     1561 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     1621 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     1681 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     1741 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     1801 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1861 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     1921 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     1981 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     2041 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     2101 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg gcagtggaga
     2161 gggcagagga agtctgctaa catgcggtga cgtcgaggag aatcctggcc cagctgagcc
     2221 ccgccaggag ttcgaagtga tggaagatca cgctgggacg tacgggttgg gggacaggaa
     2281 agatcagggg ggctacacca tgcaccaaga ccaagagggt gacacggacg ctggcctgaa
     2341 agaatctccc ctgcagaccc ccactgagga cggatctgag gaaccgggct ctgaaacctc
     2401 tgatgctaag agcactccaa cagcggaaga tgtgacagca cccttagtgg atgagggagc
     2461 tcccggcaag caggctgccg cgcagcccca cacggagatc ccagaaggaa ccacagctga
     2521 agaagcaggc attggagaca cccccagcct ggaagacgaa gctgctggtc acgtgaccca
     2581 agctcgcatg gtcagtaaaa gcaaagacgg gactggaagc gatgacaaaa aagccaaggg
     2641 ggctgatggt aaaacgaaga tcgccacacc gcggggagca gcccctccag gccagaaggg
     2701 ccaggccaac gccaccagga ttccagcaaa aaccccgccc gctccaaaga caccacccag
     2761 ctctggtgaa cctccaaaat caggggatcg cagcggctac agcagccccg gctccccagg
     2821 cactcccggc agccgctccc gcaccccgtc ccttccaacc ccacccaccc gggagcccaa
     2881 gaaggtggca gtggtccgta ctccacccaa gtcgccgtct tccgccaaga gccgcctgca
     2941 gacagccccc gtgcccatgc cagacctgaa gaatgtcaag tccaagatcg gctccactga
     3001 gaacctgaag caccagccgg gaggcgggaa ggtgcagata attaataaga agctggatct
     3061 tagcaacgtc cagtccaagt gtggctcaaa ggataatatc aaacacgtcc tgggaggcgg
     3121 cagtgtgcaa atagtctaca aaccagttga cctgagcaag gtgacctcca agtgtggctc
     3181 attaggcaac atccatcata aaccaggagg tggccaggtg gaagtaaaat ctgagaagct
     3241 tgacttcaag gacagagtcc agtcgaagat tgggtccctg gacaatatca cccacgtccc
     3301 tggcggagga aataaaaaga ttgaaaccca caagctgacc ttccgcgaga acgccaaagc
     3361 caagacagac cacggggcgg agatcgtgta caagtcgcca gtggtgtctg gggacacgtc
     3421 tccacggcat ctcagcaatg tctcctccac cggcagcatc gacatggtag actcgcccca
     3481 gctcgccacg ctagctgacg aggtgtctgc ctccctggcc aagcagggtt tgtagactag
     3541 cgatatcggt accggatccg ataatcaacc tctggattac aaaatttgtg aaagattgac
     3601 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3661 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3721 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3781 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3841 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3901 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     3961 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     4021 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     4081 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     4141 cgcctccccg catcggatcc aattcactcc tcaggtgcag gctgcctatc agaaggtggt
     4201 ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa
     4261 aaattatggg gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt
     4321 tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg
     4381 gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc
     4441 ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca
     4501 gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt
     4561 tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt
     4621 actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt
     4681 atggagatcc ctcgacctgc agcccaagct ctagagcatg gctacgtaga taagtagcat
     4741 ggcgggttaa tcattaacta caaggaaccc ctagtgatgg agttggccac tccctctctg
     4801 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     4861 cgggcggcct cagtgagcga gcgagcgcgc agctggcgta atagcgaaga ggcccgcacc
     4921 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgattccg ttgcaatggc
     4981 tggcggtaat attgttctgg atattaccag caaggccgat agtttgagtt cttctactca
     5041 ggcaagtgat gttattacta atcaaagaag tattgcgaca acggttaatt tgcgtgatgg
     5101 acagactctt ttactcggtg gcctcactga ttataaaaac acttctcagg attctggcgt
     5161 accgttcctg tctaaaatcc ctttaatcgg cctcctgttt agctcccgct ctgattctaa
     5221 cgaggaaagc acgttatacg tgctcgtcaa agcaaccata gtacgcgccc tgtagcggcg
     5281 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     5341 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     5401 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     5461 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     5521 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     5581 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     5641 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     5701 tattaacgct tacaatttaa atatttgctt atacaatctt cctgtttttg gggcttttct
     5761 gattatcaac cggggtacat atgattgaca tgctagtttt acgattaccg ttcatcgatt
     5821 ctcttgtttg ctccagactc tcaggcaatg acctgatagc ctttgtagag acctctcaaa
     5881 aatagctacc ctctccggca tgaatttatc agctagaacg gttgaatatc atattgatgg
     5941 tgatttgact gtctccggcc tttctcaccc gtttgaatct ttacctacac attactcagg
     6001 cattgcattt aaaatatatg agggttctaa aaatttttat ccttgcgttg aaataaaggc
     6061 ttctcccgca aaagtattac agggtcataa tgtttttggt acaaccgatt tagctttatg
     6121 ctctgaggct ttattgctta attttgctaa ttctttgcct tgcctgtatg atttattgga
     6181 tgttggaatc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     6241 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     6301 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     6361 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     6421 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     6481 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     6541 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     6601 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     6661 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     6721 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     6781 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     6841 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     6901 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     6961 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     7021 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     7081 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     7141 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     7201 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     7261 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     7321 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     7381 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     7441 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     7501 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     7561 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     7621 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     7681 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     7741 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     7801 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     7861 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     7921 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     7981 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     8041 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     8101 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     8161 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     8221 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     8281 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     8341 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     8401 agcgagtcag tgagcgag
//
