LOCUS       sequence-365691-        6345 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of GFP and human alpha-Synuclein with A11P and V70P 
            mutations from hSyn promoter .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6345)
  AUTHORS   Albarran E, Sun Y, Liu Y, Raju K, Dong A, Li Y, Wang S, Sudhof TC, 
            Ding JB
  TITLE     Postsynaptic synucleins mediate endocannabinoid signaling.
  JOURNAL   Nat Neurosci. 2023 Jun;26(6):997-1007. doi: 
            10.1038/s41593-023-01345-0. Epub 2023 May 29.
  PUBMED    37248337
REFERENCE   2  (bases 1 to 6345)
  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: 
            "10.1038/s41593-023-01345-0"; journalName: "Nat Neurosci"; date: 
            "2023-06"; volume: "26"; issue: "6"; pages: "997-1007"
COMMENT     Sequence Label: pAAV-hSyn-GFP-IRES-hSNCA (A11P/V70P)
FEATURES             Location/Qualifiers
     source          1..6345
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   81..210
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      742..751
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(748..750,751..753,754..1467)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(748..750,751..753,754..1464)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(793..814)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1054..1073)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1401..1422
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1495..2058
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1652..1669)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1879..1898
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      2093..2102
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2552..3140
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2605..2625)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3023..3034)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3172..3648
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3371..3390)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3688..3828
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3688..3817
                     /label=AAV2 ITR
     rep_origin      3903..4358
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3990..4009)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4200..4221
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4375..4394)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4494..4516
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4554..4572)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4640..4744
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4745..4813,4814..5605)
                     /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(4963..4982)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5776..6345,1..19)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6265..6284
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
       61 ctggcctttt gctcacatgt cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc
      121 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg
      181 gagtggccaa ctccatcact aggggttcct gcggccgcac gcgttaactg cagagggccc
      241 tgcgtatgag tgcaagtggg ttttaggacc aggatgaggc ggggtggggg tgcctacctg
      301 acgaccgacc ccgacccact ggacaagcac ccaaccccca ttccccaaat tgcgcatccc
      361 ctatcagaga gggggagggg aaacaggatg cggcgaggcg cgtgcgcact gccagcttca
      421 gcaccgcgga cagtgccttc gcccccgcct ggcggcgcgc gccaccgccg cctcagcact
      481 gaaggcgcgc tgacgtcact cgccggtccc ccgcaaactc cccttcccgg ccaccttggt
      541 cgcgtccgcg ccgccgccgg cccagccgga ccgcaccacg cgaggcgcga gatagggggg
      601 cacgggcgcg accatctgcg ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg
      661 ggcagcggag gagtcgtgtc gtgcctgaga gcgcgtccgg agctattagc aatatgaaac
      721 ctcttaggcg cgccaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
      781 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
      841 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
      901 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
      961 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     1021 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     1081 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     1141 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     1201 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     1261 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     1321 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
     1381 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     1441 ctcggcatgg acgagctgta caagtaaagc cagtgtggtg gtacgtagga attctccctc
     1501 ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg
     1561 ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc
     1621 ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg
     1681 aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg
     1741 accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca
     1801 cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata
     1861 gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc
     1921 cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt
     1981 gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt
     2041 tgaaaaacac gatgataatg gtaccgcggg cccgggatcc tctagagtcg acgccaccat
     2101 ggatgtattc atgaaaggac tttcaaagcc caaggaggga gtggtggctg ctgctgagaa
     2161 aaccaaacag ggtgtggcag aagcagcagg aaagacaaaa gagggtgttc tctatgtagg
     2221 ctccaaaacc aaggagggag tggtgcatgg tgtggcaaca gtggctgaga agaccaaaga
     2281 gcaagtgaca aatgttggag gagcaccggt gacgggtgtg acagcagtag cccagaagac
     2341 agtggaggga gcagggagca ttgcagcagc cactggcttt gtcaaaaagg accagttggg
     2401 caagaatgaa gaaggagccc cacaggaagg aattctggaa gatatgcctg tggatcctga
     2461 caatgaggct tatgaaatgc cttctgagga agggtatcaa gactacgaac ctgaagccta
     2521 agctagcgaa ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga
     2581 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     2641 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     2701 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     2761 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     2821 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     2881 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     2941 ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg
     3001 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     3061 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     3121 cctttgggcc gcctccccgc atcgataccg agcgctgctc gagagatcta cgggtggcat
     3181 ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc
     3241 agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat
     3301 attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg
     3361 gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca
     3421 atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc
     3481 caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc
     3541 atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc
     3601 aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttgtag
     3661 gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
     3721 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     3781 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     3841 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     3901 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3961 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4021 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4081 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     4141 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4201 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     4261 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     4321 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     4381 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     4441 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     4501 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     4561 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     4621 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4681 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4741 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4801 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4861 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4921 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4981 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5041 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5101 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5161 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5221 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5281 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5341 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5401 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5461 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5521 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5581 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     5641 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5701 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5761 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5821 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5881 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5941 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6001 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6061 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6121 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6181 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6241 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6301 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcagg
//
