LOCUS       sequence-365694-        6345 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of GFP and human alpha-Synuclein with S129D mutation 
            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 (S129D)
FEATURES             Location/Qualifiers
     source          1..6345
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      593..602
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(599..601,602..604,605..1318)
                     /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(599..601,602..604,605..1315)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(644..665)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(905..924)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1252..1273
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1346..1909
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1503..1520)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1730..1749
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      1944..1953
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2403..2991
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2456..2476)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2874..2885)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3023..3499
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3222..3241)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3539..3679
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3539..3668
                     /label=AAV2 ITR
     rep_origin      3754..4209
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3841..3860)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4051..4072
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4226..4245)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4345..4367
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4405..4423)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4491..4595
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4596..4664,4665..5456)
                     /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(4814..4833)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5627..6215
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6116..6135
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6277..6345,1..61)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc
       61 tgcggccgca cgcgttaact gcagagggcc ctgcgtatga gtgcaagtgg gttttaggac
      121 caggatgagg cggggtgggg gtgcctacct gacgaccgac cccgacccac tggacaagca
      181 cccaaccccc attccccaaa ttgcgcatcc cctatcagag agggggaggg gaaacaggat
      241 gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg acagtgcctt cgcccccgcc
      301 tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg ctgacgtcac tcgccggtcc
      361 cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc gccgccgccg gcccagccgg
      421 accgcaccac gcgaggcgcg agataggggg gcacgggcgc gaccatctgc gctgcggcgc
      481 cggcgactca gcgctgcctc agtctgcggt gggcagcgga ggagtcgtgt cgtgcctgag
      541 agcgcgtccg gagctattag caatatgaaa cctcttaggc gcgccaccgg tcgccaccat
      601 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
      661 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
      721 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
      781 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
      841 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
      901 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
      961 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1021 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     1081 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     1141 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1201 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1261 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaag
     1321 ccagtgtggt ggtacgtagg aattctccct cccccctaac gttactggcc gaagccgctt
     1381 ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg
     1441 caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc
     1501 ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga
     1561 agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc
     1621 tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc
     1681 acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc
     1741 aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga
     1801 tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg
     1861 ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat ggtaccgcgg
     1921 gcccgggatc ctctagagtc gacgccacca tggatgtatt catgaaagga ctttcaaagg
     1981 ccaaggaggg agttgtggct gctgctgaga aaaccaaaca gggtgtggca gaagcagcag
     2041 gaaagacaaa agagggtgtt ctctatgtag gctccaaaac caaggaggga gtggtgcatg
     2101 gtgtggcaac agtggctgag aagaccaaag agcaagtgac aaatgttgga ggagcagtgg
     2161 tgacgggtgt gacagcagta gcccagaaga cagtggaggg agcagggagc attgcagcag
     2221 ccactggctt tgtcaaaaag gaccagttgg gcaagaatga agaaggagcc ccacaggaag
     2281 gaattctgga agatatgcct gtggatcctg acaatgaggc ttatgaaatg cctgatgagg
     2341 aagggtatca agactacgaa cctgaagcct aagctagcga attcgatatc aagcttatcg
     2401 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     2461 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     2521 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     2581 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     2641 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     2701 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     2761 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     2821 tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     2881 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     2941 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc
     3001 gagcgctgct cgagagatct acgggtggca tccctgtgac ccctccccag tgcctctcct
     3061 ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat
     3121 cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag
     3181 caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg
     3241 gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct
     3301 cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt
     3361 tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta
     3421 atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac
     3481 tgctcccttc cctgtccttc tgattttgta ggtaaccacg tgcggaccga gcggccgcag
     3541 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     3601 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     3661 gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     3721 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
     3781 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg
     3841 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     3901 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3961 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     4021 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     4081 tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt
     4141 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     4201 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     4261 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     4321 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     4381 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     4441 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4501 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4561 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     4621 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     4681 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     4741 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     4801 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     4861 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     4921 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     4981 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     5041 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     5101 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     5161 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     5221 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     5281 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     5341 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     5401 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     5461 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5521 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5581 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5641 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5701 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5761 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5821 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5881 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5941 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     6001 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     6061 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     6121 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6181 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6241 ggttcctggc cttttgctgg ccttttgctc acatgtcctg caggcagctg cgcgctcgct
     6301 cgctcactga ggccgcccgg gcgtcgggcg acctttggtc gcccg
//
