LOCUS       sequence-365689-        6210 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of GFP and C-terminal truncated (1-95 amino acid) 
            mouse alpha-Synuclein from hSyn promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6210)
  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 6210)
  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-mSNCA(1-95)
FEATURES             Location/Qualifiers
     source          1..6210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      659..668
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(665..667,668..670,671..1384)
                     /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(665..667,668..670,671..1381)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(710..731)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(971..990)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1318..1339
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1412..1975
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1569..1586)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1796..1815
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      2010..2019
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2334..2922
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2387..2407)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2805..2816)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2954..3430
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3153..3172)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3470..3610
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3470..3599
                     /label=AAV2 ITR
     rep_origin      3685..4140
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3772..3791)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3982..4003
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4157..4176)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4276..4298
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4336..4354)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4422..4526
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4527..4595,4596..5387)
                     /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(4745..4764)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5558..6146
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6047..6066
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6208..6210,1..127)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt
       61 cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg
      121 ggttcctgcg gccgcacgcg ttaactgcag agggccctgc gtatgagtgc aagtgggttt
      181 taggaccagg atgaggcggg gtgggggtgc ctacctgacg accgaccccg acccactgga
      241 caagcaccca acccccattc cccaaattgc gcatccccta tcagagaggg ggaggggaaa
      301 caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca ccgcggacag tgccttcgcc
      361 cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa ggcgcgctga cgtcactcgc
      421 cggtcccccg caaactcccc ttcccggcca ccttggtcgc gtccgcgccg ccgccggccc
      481 agccggaccg caccacgcga ggcgcgagat aggggggcac gggcgcgacc atctgcgctg
      541 cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc agcggaggag tcgtgtcgtg
      601 cctgagagcg cgtccggagc tattagcaat atgaaacctc ttaggcgcgc caccggtcgc
      661 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      721 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
      781 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
      841 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
      901 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
      961 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1021 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1081 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1141 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1201 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1261 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1321 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1381 gtaaagccag tgtggtggta cgtaggaatt ctccctcccc cctaacgtta ctggccgaag
     1441 ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc
     1501 ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg
     1561 tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc
     1621 tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc
     1681 cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa
     1741 ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct
     1801 ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg
     1861 atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg
     1921 tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatggta
     1981 ccgcgggccc gggatcctct agagtcgacg ccaccatgga tgtgttcatg aaaggacttt
     2041 caaaggccaa ggagggagtt gtggctgctg ctgagaaaac caagcagggt gtggcagagg
     2101 cagctggaaa gacaaaagag ggagtcctct atgtaggttc caaaactaag gaaggagtgg
     2161 ttcatggagt gacaacagtg gctgagaaga ccaaagagca agtgacaaat gttggaggag
     2221 cagtggtgac tggtgtgaca gcagtcgctc agaagacagt ggagggagct gggaatatag
     2281 ctgctgccac tggctttgtc taagctagcg aattcgatat caagcttatc gataatcaac
     2341 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     2401 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     2461 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     2521 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     2581 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     2641 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     2701 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctatg
     2761 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     2821 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     2881 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgagcgctgc
     2941 tcgagagatc tacgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga
     3001 agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc
     3061 tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca
     3121 agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt
     3181 ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca
     3241 gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt
     3301 ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt
     3361 gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt
     3421 ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg agcggccgca ggaaccccta
     3481 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     3541 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     3601 tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3661 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3721 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg
     3781 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3841 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3901 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3961 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta
     4021 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa
     4081 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     4141 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     4201 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     4261 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     4321 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     4381 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     4441 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     4501 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     4561 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     4621 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     4681 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     4741 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     4801 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     4861 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     4921 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     4981 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     5041 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     5101 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     5161 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     5221 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     5281 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     5341 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     5401 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5461 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5521 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5581 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5641 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5701 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5761 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5821 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5881 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5941 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6001 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6061 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6121 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6181 ccttttgctg gccttttgct cacatgtcct
//
