LOCUS       sequence-469454-        6690 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of Venus-P2A-Tau-RING from CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6690)
  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 6690)
  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 PHP.eB-CAG-Venus-P2A-Tau-RING
FEATURES             Location/Qualifiers
     source          1..6690
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   179..308
                     /label=AAV2 ITR
     rep_origin      380..835
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(467..486)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     677..698
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        861..965
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(966..1034,1035..1826)
                     /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(1184..1203)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1997..2585
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2486..2505
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2964..3085
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3001..3020)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3055..3074
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    3128..3716
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3234..3245
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3643..3663
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5178..5234)
                     /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(5250..5960,5961..5963,5964..5966))
                     /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(5292..5313)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     5641..5660
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5900..5921
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5979..6001)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(6280..6558)
                     /label=chicken beta-actin promoter
ORIGIN
        1 gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc
       61 aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac gtcaatggta
      121 ctcttcctag agcatggcta cgtagataag tagcatggcg ggttaatcat taactacaag
      181 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
      241 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
      301 gcgcgcagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
      361 agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
      421 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
      481 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
      541 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
      601 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
      661 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
      721 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
      781 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
      841 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
      901 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
      961 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     1021 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     1081 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     1141 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     1201 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     1261 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     1321 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     1381 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     1441 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     1501 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     1561 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     1621 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     1681 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     1741 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     1801 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1861 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     1921 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1981 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     2041 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     2101 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     2161 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     2221 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     2281 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     2341 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     2401 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     2461 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     2521 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     2581 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     2641 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     2701 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgagcgag
     2761 cgcgcagaga gggagtggcc aactccatca ctaggggttc cttgtagtta atgattaacc
     2821 cgccatgcta cttatctacg tagccatgct ctaggaagat ccttatcgat tttaccacat
     2881 ttgtagaggt tttacttgct ttaaaaaacc tcccacatct ccccctgaac ctgaaacata
     2941 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     3001 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3061 tgtccaaact catcaatgta tcttatcatg tctgctcgaa gcggccgccc gggtcgacgg
     3121 tatcgatgcg gggaggcggc ccaaagggag atccgactcg tctgagggcg aaggcgaaga
     3181 cgcggaagag gccgcagagc cggcagcagg ccgcgggaag gaaggtccgc tggattgagg
     3241 gccgaaggga cgtagcagaa ggacgtcccg cgcagaatcc aggtggcaac acaggcgagc
     3301 agccaaggaa aggacgatga tttccccgac aacaccacgg aattgtcagt gcccaacagc
     3361 cgagcccctg tccagcagcg ggcaaggcag gcggcgatga gttccgccgt ggcaataggg
     3421 agggggaaag cgaaagtccc ggaaaggagc tgacaggtgg tggcaatgcc ccaaccagtg
     3481 ggggttgcgt cagcaaacac agtgcacacc acgccacgtt gcctgacaac gggccacaac
     3541 tcctcataaa gagacagcaa ccaggattta tacaaggagg agaaaatgaa agccatacgg
     3601 gaagcaatag catgatacaa aggcattaaa gcagcgtatc cacatagcgt aaaaggagca
     3661 acatagttaa gaataccagt caatctttca caaattttgt aatccagagg ttgattatcg
     3721 ataagcttga tatcttactc tctggcctcc tggctgattt ctttaaggtt gttcaccatg
     3781 ttggctagct gtcgattggg ccggagattc ttgagcagaa agcgctgccg gcacacagga
     3841 cagacgctgc ccccaccttt cccaacctga gagatgcatt cctggcagaa gctgtggcca
     3901 cactcgatgc tcacaggctc cacgaagggg tccaggcaga tagggcatgt gacctcctcc
     3961 cacatcattg tcaagcgtgc tgctgaagcc atgcctgagc cacctccgga tccgaattcc
     4021 aaaccctgct tggccaggga ggcagacacc tcgtcagcta gcgtggcgag ctggggcgag
     4081 tctaccatgt cgatgctgcc ggtggaggag acattgctga gatgccgtgg agacgtgtcc
     4141 ccagacacca ctggcgactt gtacacgatc tccgccccgt ggtctgtctt ggctttggcg
     4201 ttctcgcgga aggtcagctt gtgggtttca atctttttat ttcctccgcc agggacgtgg
     4261 gtgatattgt ccagggaccc aatcttcgac tggactctgt ccttgaagtc aagcttctca
     4321 gattttactt ccacctggcc acctcctggt ttatgatgga tgttgcctaa tgagccacac
     4381 ttggaggtca ccttgctcag gtcaactggt ttgtagacta tttgcacact gccgcctcca
     4441 ctgacgtgtt tgatattatc ctttgagcca cacttggact ggacgttgct aagatccagc
     4501 ttcttattaa ttatctgcac cttcccgcct cccggctggt gcttcaggtt ctcagtggag
     4561 ccgatcttgg acttgacatt cttcaggtct ggcatgggca cgggggctgt ctgcaggcgg
     4621 ctcttggcgg aagacggcga cttgggtgga gtacggacca ctgccacctt cttgggctcc
     4681 cgggtgggtg gggttggaag ggacggggtg cgggagcggc tgccgggagt gcctggggag
     4741 ccggggctgc tgtagccgct gcgatcccct gattttggag gttcaccaga gctgggtggt
     4801 gtctttggag cgggcggggt ttttgctgga atcctggtgg cgttggcctg gcccttctgg
     4861 cctggagggg ctgctccccg cggtgtggcg atcttcgttt taccatcagc ccccttggct
     4921 tttttgtcat cgcttccagt cccgtctttg cttttactga ccatgcgagc ttgggtcacg
     4981 tgaccagcag cttcgtcttc caggctgggg gtgtctccaa tgcctgcttc ttcagctttc
     5041 aggccagcgt ccgtgtcacc ctcttggtct tggtgcatgg tgtagccccc ctgatctttc
     5101 ctgtccccca acccgtacgt cccagcgtga tcttccatca cttcgaactc ctggcggggc
     5161 tcagccatct taattaaagg tccagggttc tcctccacgt ctccagcctg cttcagcagg
     5221 ctgaagttag tagctccgct tccgaattcc ttgtacagct cgtccatgcc gagagtgatc
     5281 ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg
     5341 ctcagggcgg actggtagct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg
     5401 atgggggtgt tctgctggta gtggtcggcg agctgcacgc cgccgtcctc gatgttgtgg
     5461 cggatcttga agttggcctt gatgccgttc ttctgcttgt cggcggtgat atagacgttg
     5521 tggctgttgt agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg
     5581 atgcccttca gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg
     5641 gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc
     5701 atggcggact tgaagaagtc gtgctgcttc atgtggtcgg ggtagcgggc gaagcactgc
     5761 aggccgtagc ccagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg
     5821 cagatcagct tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg
     5881 ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg
     5941 aacagctcct cgcccttgct caccattcta gagccgccgg tcacacgcca gaagccgaac
     6001 cccgccctgc cccgtccccc ccgaaggcag ccgtcccccc gcggacagcc ccgagccgct
     6061 ccccccgcac aaagggccct cccggagccc ctcaaggctt tcacgcagcc acagaaaaga
     6121 aacaagccgt cattaaacca agcgctaatt acagcccgga ggagaagggc cgtcccgccc
     6181 gctcacctgt gggagtaacg cggtcagtca gagccggggc gggcggcgcg aggcggcggc
     6241 ggagcggggc acggggcgaa ggcagcgcgc agcgactccc gcccgccgcg cgcttcgctt
     6301 tttatagggc cgccgccgcc gccgcctcgc cataaaagga aactttcgga gcgcgccgct
     6361 ctgattggct gccgccgcac ctctccgcct cgccccgccc cgcccctcgc cccgccccgc
     6421 cccgcccggc gcgccccccc cccccccccc ccgcccccat cgctgcacaa aataattaaa
     6481 aaataaataa atacaaaatt gggggtgggg agggggggga gatggggaga gtgaagcaga
     6541 acgtggggct cacctcgacc atggtaatag cgatgactaa tacgtagatg tactgccaag
     6601 taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat
     6661 tgacgtcaat agggggcgta cttggcatat
//
