LOCUS       sequence-390731-        6345 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of GFP and human alpha-Synuclein with A30P 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 (A30P)
FEATURES             Location/Qualifiers
     source          1..6345
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      662..671
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(668..670,671..673,674..1387)
                     /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(668..670,671..673,674..1384)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(713..734)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(974..993)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1321..1342
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1415..1978
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1572..1589)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1799..1818
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      2013..2022
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2472..3060
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2525..2545)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2943..2954)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3092..3568
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3291..3310)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3608..3748
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3608..3737
                     /label=AAV2 ITR
     rep_origin      3823..4278
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3910..3929)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4120..4141
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4295..4314)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4414..4436
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4474..4492)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4560..4664
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4665..4733,4734..5525)
                     /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(4883..4902)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5696..6284
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6185..6204
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttaactg cagagggccc tgcgtatgag tgcaagtggg
      181 ttttaggacc aggatgaggc ggggtggggg tgcctacctg acgaccgacc ccgacccact
      241 ggacaagcac ccaaccccca ttccccaaat tgcgcatccc ctatcagaga gggggagggg
      301 aaacaggatg cggcgaggcg cgtgcgcact gccagcttca gcaccgcgga cagtgccttc
      361 gcccccgcct ggcggcgcgc gccaccgccg cctcagcact gaaggcgcgc tgacgtcact
      421 cgccggtccc ccgcaaactc cccttcccgg ccaccttggt cgcgtccgcg ccgccgccgg
      481 cccagccgga ccgcaccacg cgaggcgcga gatagggggg cacgggcgcg accatctgcg
      541 ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg ggcagcggag gagtcgtgtc
      601 gtgcctgaga gcgcgtccgg agctattagc aatatgaaac ctcttaggcg cgccaccggt
      661 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
      721 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
      781 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
      841 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
      901 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
      961 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     1021 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     1081 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     1141 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     1201 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1261 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     1321 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1381 caagtaaagc cagtgtggtg gtacgtagga attctccctc ccccctaacg ttactggccg
     1441 aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc
     1501 gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag
     1561 gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt
     1621 tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa
     1681 ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc
     1741 aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg
     1801 gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat
     1861 gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa
     1921 acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataatg
     1981 gtaccgcggg cccgggatcc tctagagtcg acgccaccat ggatgtattc atgaaaggac
     2041 tttcaaaggc caaggaggga gttgtggctg ctgctgagaa aaccaaacag ggtgtggcag
     2101 aagcaccagg aaagacaaaa gagggtgttc tctatgtagg ctccaaaacc aaggagggag
     2161 tggtgcatgg tgtggcaaca gtggctgaga agaccaaaga gcaagtgaca aatgttggag
     2221 gagcagtggt gacgggtgtg acagcagtag cccagaagac agtggaggga gcagggagca
     2281 ttgcagcagc cactggcttt gtcaaaaagg accagttggg caagaatgaa gaaggagccc
     2341 cacaggaagg aattctggaa gatatgcctg tggatcctga caatgaggct tatgaaatgc
     2401 cttctgagga agggtatcaa gactacgaac ctgaagccta agctagcgaa ttcgatatca
     2461 agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     2521 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     2581 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     2641 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     2701 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     2761 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     2821 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     2881 cttggctgct cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     2941 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     3001 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     3061 atcgataccg agcgctgctc gagagatcta cgggtggcat ccctgtgacc cctccccagt
     3121 gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc taataaaatt
     3181 aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt ggaggggggt
     3241 ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg tctattggga
     3301 accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct cctgggttca
     3361 agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca tgaccaggct
     3421 cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca ggctggtctc
     3481 caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg gattacaggc
     3541 gtgaaccact gctcccttcc ctgtccttct gattttgtag gtaaccacgt gcggaccgag
     3601 cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     3661 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
     3721 agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat
     3781 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
     3841 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct
     3901 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3961 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4021 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4081 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4141 gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
     4201 cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     4261 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     4321 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     4381 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     4441 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     4501 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     4561 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4621 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     4681 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4741 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     4801 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     4861 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     4921 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     4981 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5041 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     5101 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5161 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5221 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5281 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5341 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5401 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5461 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5521 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     5581 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5641 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5701 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5761 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5821 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     5881 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5941 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6001 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6061 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6121 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6181 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6241 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6301 cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
