LOCUS       sequence-469455-        5302 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of Venus from CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5302)
  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 5302)
  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
FEATURES             Location/Qualifiers
     source          1..5302
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     26..45
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     285..306
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(364..386)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     repeat_region   1378..1507
                     /label=AAV2 ITR
     rep_origin      1579..2034
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1666..1685)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1876..1897
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2060..2164
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2165..2233,2234..3025)
                     /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(2383..2402)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3196..3784
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3685..3704
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4163..4284
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4200..4219)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4254..4273
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    4327..4915
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             4433..4444
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4842..4862
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(4934..5302,1..345,346..348,349..351))
                     /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(4979..5000)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gccctcgaac ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga agaagatggt
       61 gcgctcctgg acgtagcctt cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg
      121 gtcggggtag cgggcgaagc actgcaggcc gtagcccagg gtggtcacga gggtgggcca
      181 gggcacgggc agcttgccgg tggtgcagat cagcttcagg gtcagcttgc cgtaggtggc
      241 atcgccctcg ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc cgtccagctc
      301 gaccaggatg ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca ttctagagcc
      361 gccggtcaca cgccagaagc cgaaccccgc cctgccccgt cccccccgaa ggcagccgtc
      421 cccccgcgga cagccccgag ccgctccccc cgcacaaagg gccctcccgg agcccctcaa
      481 ggctttcacg cagccacaga aaagaaacaa gccgtcatta aaccaagcgc taattacagc
      541 ccggaggaga agggccgtcc cgcccgctca cctgtgggag taacgcggtc agtcagagcc
      601 ggggcgggcg gcgcgaggcg gcggcggagc ggggcacggg gcgaaggcag cgcgcagcga
      661 ctcccgcccg ccgcgcgctt cgctttttat agggccgccg ccgccgccgc ctcgccataa
      721 aaggaaactt tcggagcgcg ccgctctgat tggctgccgc cgcacctctc cgcctcgccc
      781 cgccccgccc ctcgccccgc cccgccccgc ctggcgcgcc cccccccccc cccccccccc
      841 caaaagaagc cttactcggc gccccgcccc gcggctgcgc cgcccccccc cccccccccc
      901 cccccccccc cccccccccc cccccccccg ccccccctgc gggccccccc cccccccccc
      961 cgcccccatc gctgcacaaa ataattaaaa aataaataaa tacaaaattg ggggtgggga
     1021 ggggggggag atggggagag tgaagcagaa cgtggggctc acctcgacca tggtaatagc
     1081 gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc
     1141 ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg
     1201 atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca
     1261 atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg tcaatggtac
     1321 tcttcctaga gcatggctac gtagataagt agcatggcgg gttaatcatt aactacaagg
     1381 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     1441 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     1501 cgcgcagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     1561 gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     1621 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     1681 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     1741 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     1801 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     1861 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     1921 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     1981 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     2041 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     2101 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     2161 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     2221 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     2281 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     2341 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     2401 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     2461 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     2521 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     2581 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     2641 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     2701 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     2761 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     2821 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     2881 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     2941 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     3001 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     3061 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     3121 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3181 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3241 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3301 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3361 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3421 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3481 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3541 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3601 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3661 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3721 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3781 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3841 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     3901 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgagcgagc
     3961 gcgcagagag ggagtggcca actccatcac taggggttcc ttgtagttaa tgattaaccc
     4021 gccatgctac ttatctacgt agccatgctc taggaagatc cttatcgatt ttaccacatt
     4081 tgtagaggtt ttacttgctt taaaaaacct cccacatctc cccctgaacc tgaaacataa
     4141 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     4201 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     4261 gtccaaactc atcaatgtat cttatcatgt ctgctcgaag cggccgcccg ggtcgacggt
     4321 atcgatgcgg ggaggcggcc caaagggaga tccgactcgt ctgagggcga aggcgaagac
     4381 gcggaagagg ccgcagagcc ggcagcaggc cgcgggaagg aaggtccgct ggattgaggg
     4441 ccgaagggac gtagcagaag gacgtcccgc gcagaatcca ggtggcaaca caggcgagca
     4501 gccaaggaaa ggacgatgat ttccccgaca acaccacgga attgtcagtg cccaacagcc
     4561 gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag ttccgccgtg gcaataggga
     4621 gggggaaagc gaaagtcccg gaaaggagct gacaggtggt ggcaatgccc caaccagtgg
     4681 gggttgcgtc agcaaacaca gtgcacacca cgccacgttg cctgacaacg ggccacaact
     4741 cctcataaag agacagcaac caggatttat acaaggagga gaaaatgaaa gccatacggg
     4801 aagcaatagc atgatacaaa ggcattaaag cagcgtatcc acatagcgta aaaggagcaa
     4861 catagttaag aataccagtc aatctttcac aaattttgta atccagaggt tgattatcga
     4921 taagcttgat atcttacttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca
     4981 cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc agggcggact
     5041 ggtagctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct
     5101 gctggtagtg gtcggcgagc tgcacgccgc cgtcctcgat gttgtggcgg atcttgaagt
     5161 tggccttgat gccgttcttc tgcttgtcgg cggtgatata gacgttgtgg ctgttgtagt
     5221 tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct
     5281 cgatgcggtt caccagggtg tc
//
