LOCUS       sequence-95968-e        5835 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An AAV packaging vector that expresses wildtype alpha-synuclein 
            under control of the EF1a promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5835)
  AUTHORS   Savolainen MH, Richie CT, Harvey BK, Mannisto PT, Maguire-Zeiss KA, 
            Myohanen TT
  TITLE     The beneficial effect of a prolyl oligopeptidase inhibitor, 
            KYP-2047, on alpha-synuclein clearance and autophagy in A30P 
            transgenic mouse.
  JOURNAL   Neurobiol Dis. 2014 Aug;68:1-15. doi: 10.1016/j.nbd.2014.04.003. 
            Epub 2014 Apr 16.
  PUBMED    24746855
REFERENCE   2  (bases 1 to 5835)
  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: "Neurobiol 
            Dis."; date: "2014-08"; volume: "68:1-15. doi"; pages: " 
            10.1016/j.nbd.2014.04.003. Epub 2014 Apr 16"
COMMENT     Sequence Label: pOTTC293 - pAAV EF1a V5-synuclein (WT)
FEATURES             Location/Qualifiers
     source          1..5835
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        242..1419
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          473..1410
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1367..1387
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1435..1476
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    1962..2550
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2015..2035)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2433..2444)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2582..3058
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2781..2800)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3098..3238
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3098..3227
                     /label=AAV2 ITR
     rep_origin      3313..3768
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3400..3419)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3610..3631
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3785..3804)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3904..3926
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3964..3982)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4050..4154
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4155..4223,4224..5015)
                     /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(4373..4392)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5186..5774
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5675..5694
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt
      181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
      241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc catgggtaag
     1441 cctatcccta accctctcct cggtctcgat tctacggcta gcatggatgt attcatgaaa
     1501 ggactttcaa aggccaagga gggagttgtg gctgctgctg agaaaaccaa acagggtgtg
     1561 gcagaagcag caggaaagac aaaagagggt gttctctatg taggctccaa aaccaaggag
     1621 ggagtggtgc atggtgtggc aacagtggct gagaagacca aagagcaagt gacaaatgtt
     1681 ggaggagcag tggtgacggg tgtgacagca gtagcccaga agacagtgga gggagcaggg
     1741 agcattgcag cagccactgg ctttgtcaaa aaggaccagt tgggcaagaa tgaagaagga
     1801 gccccacagg aaggaattct ggaagatatg cctgtggatc ctgacaatga ggcttatgaa
     1861 atgccttctg aggaagggta tcaagactac gaacctgaag cctaagaaat atctttgctc
     1921 ccagtttctt gagatctgaa ttcgatatca agcttatcga taatcaacct ctggattaca
     1981 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     2041 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     2101 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     2161 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     2221 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     2281 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     2341 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga
     2401 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     2461 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     2521 gtcggatctc cctttgggcc gcctccccgc atcgataccg agcgctgctc gagagatcta
     2581 cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc
     2641 agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc
     2701 cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac
     2761 aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg
     2821 gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt
     2881 gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg
     2941 gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc
     3001 ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct
     3061 gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg
     3121 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     3181 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     3241 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     3301 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3361 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     3421 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     3481 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     3541 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3601 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcgggctatt
     3661 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     3721 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     3781 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     3841 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     3901 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     3961 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     4021 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4081 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4141 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4201 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4261 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4321 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4381 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4441 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     4501 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4561 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     4621 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4681 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     4741 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4801 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4861 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     4921 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4981 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     5041 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5101 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5161 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5221 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5281 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     5341 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5401 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5461 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5521 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5581 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5641 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5701 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5761 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5821 cttttgctca catgt
//
