LOCUS       sequence-379299-        6223 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-vector for optogenetic long-term silencing of neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6223)
  AUTHORS   Rodriguez-Rozada S, Wietek J, Tenedini F, Sauter K, Dhiman N, 
            Hegemann P, Soba P, Wiegert JS
  TITLE     Aion is a bistable anion-conducting channelrhodopsin that provides 
            temporally extended and reversible neuronal silencing.
  JOURNAL   Commun Biol. 2022 Jul 9;5(1):687. doi: 10.1038/s42003-022-03636-x.
  PUBMED    35810216
REFERENCE   2  (bases 1 to 6223)
  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: "Commun 
            Biol."; date: "2022-07-9"; volume: "5(1):687. doi"; pages: " 
            10.1038/s42003-022-03636-x"
COMMENT     Sequence Label: hSyn-somAion-citrine
FEATURES             Location/Qualifiers
     source          1..6223
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     104..123
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   282..411
                     /label=AAV2 ITR
     promoter        511..958
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      978..987
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1926..1928,1929..2639)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2229..2248)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2856..3444
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2909..2929)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3327..3338)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3485..3692
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3862..3991
                     /label=AAV2 ITR
     rep_origin      complement(4228..4816)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4308..4327)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4987..5778,5779..5847))
                     /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     5610..5629
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5848..5952)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(5978..6223,1..210))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6115..6136)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
       61 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
      121 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
      181 ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt
      241 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctgcgcgct cgctcgctca
      301 ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga
      361 gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc ttgtagttaa
      421 tgattaaccc gccatgctac ttatctacgt agccatgctc taggaagatc gtaggttaat
      481 taatctagac tgcagagggc cctgcgtatg agtgcaagtg ggttttagga ccaggatgag
      541 gcggggtggg ggtgcctacc tgacgaccga ccccgaccca ctggacaagc acccaacccc
      601 cattccccaa attgcgcatc ccctatcaga gagggggagg ggaaacagga tgcggcgagg
      661 cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct tcgcccccgc ctggcggcgc
      721 gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca ctcgccggtc ccccgcaaac
      781 tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc ggcccagccg gaccgcacca
      841 cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg cgctgcggcg ccggcgactc
      901 agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg tcgtgcctga gagcgcagtc
      961 gaattcaagc tgctagcgcc accatggact atggcggcgc tttgtctgcc gtcggactct
     1021 tccaaacctc atacactctt gagaacaatg gttctgttat ttgcatcccg aacaacggcc
     1081 agtgcttctg cttggcttgg cttaaatcca acggaacaaa tgccgagaag ttggctgcca
     1141 acattctgca gtggatttct tttgcgcttt cagcgctctg cctgatgttc tacggctacc
     1201 agacctggaa gtctacttgc ggctgggaga acatttacgt ggccacgatc cagatgatca
     1261 agttcatcat cgagtatttc catagctttg acgaacctgc ggtgatctac tcatccaacg
     1321 gcaacaagac ccggtggctt cgttacgcga gctggctgct gaccgcccct gtcattctta
     1381 tccacctgag caaccttacg ggtctggcga acgactataa caagcgtacc atgggtctgc
     1441 tggtgtcatg catcggcggc atcgtgtggg ccaccacggc cgcgctgtcc aagggatacg
     1501 tccgtgtcat tttcttcctg atgggcctgt gctacggcat ctacacattc ttcaacgcag
     1561 ccaaggtcta cattgaggcg taccacaccg tgccgaaggg ccggtgtcgc caggtggtga
     1621 ctggcatggc ttggctcttc ttcgtatcat ggggtatgtt ccccatcctg ttcatcctcg
     1681 gccccgaggg cttcggcgtc ctgagccggt acggctccaa cgtcggccac accatcattg
     1741 acctgatgtc gaagcagtgc tggggtctgc tcggccacta cctgcgcgtg ctgatccaca
     1801 gccatatcct catccacggc gacattcgca agaccaccaa attgaacatt ggtggcactg
     1861 agattgaggt cgagacgctg gtggaggacg aggccgaggc tggcgcggtc gcggcactcg
     1921 aggtggtgag caagggcgag gagctgttca ccggcgtggt gcccatcctg gtggagctgg
     1981 acggcgacgt gaacggccac aagttcagcg tgagcggcga gggcgagggc gacgccacct
     2041 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     2101 ccctggtgac caccttcggc tacggcctga tgtgcttcgc ccgctacccc gaccacatga
     2161 agcagcacga cttcttcaag agcgccatgc ccgagggcta cgtgcaggag cgcaccatct
     2221 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     2281 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctgggcc
     2341 acaagctgga gtacaactac aacagccaca acgtgtacat catggccgac aagcagaaga
     2401 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctgg
     2461 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     2521 actacctgag ctaccagagc gccctgagca aggaccccaa cgagaagcgc gaccacatgg
     2581 tgctgctgga gttcgtgacc gccgccggca tcaccctggg catggacgag ctgtacaagc
     2641 agagccagcc catcctgaac accaaggaga tggcccccca gagcaagccc cccgaggagc
     2701 tggagatgag cagcatgccc agccccgtgg cccccctgcc cgcccgcacc gagggcgtga
     2761 tcgacatgcg cagcatgagc agcatcgaca gcttcatcag ctgcgccacc gacttccccg
     2821 aggccacccg cttctgatct agaaagctta tcgataatca acctctggat tacaaaattt
     2881 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     2941 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     3001 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     3061 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     3121 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     3181 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     3241 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     3301 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     3361 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     3421 tctccctttg ggccgcctcc ccgcatcgat accgtcgacc cgggcggccg ctcataaagc
     3481 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     3541 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     3601 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     3661 gattgggaag acaatagcag gcatgctggg gatgcggtcc ggactgtaca attgcattca
     3721 ttttatgttt caggttcagg gggagatgtg ggaggttttt taaagcaagt aaaacctcta
     3781 caaatgtggt aaaatcgata aggatcttcc tagagcatgg ctacgtagat aagtagcatg
     3841 gcgggttaat cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc
     3901 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     3961 gggcggcctc agtgagcgag cgagcgcgca gctgcattaa tgaatcggcc aacgcgcggg
     4021 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4081 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4141 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4201 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4261 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4321 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4381 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4441 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4501 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4561 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4621 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4681 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4741 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4801 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4861 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4921 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4981 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5041 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5101 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5161 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5221 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5281 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5341 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5401 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5461 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5521 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5581 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5641 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5701 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5761 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5821 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5881 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5941 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg
     6001 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     6061 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     6121 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     6181 tatcagggcg atggcccact acgtgaacca tcaccctaat caa
//
