LOCUS       sequence-191035-        5843 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetically encoded voltage indicator (GEVI) ASAP2s in AAV vector 
            for neuron-specific expression using the promoter hSyn1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5843)
  AUTHORS   Chamberland S, Yang HH, Pan MM, Evans SW, Guan S, Chavarha M, Yang 
            Y, Salesse C, Wu H, Wu JC, Clandinin TR, Toth K, Lin MZ, St-Pierre F
  TITLE     Fast two-photon imaging of subcellular voltage dynamics in neuronal 
            tissue with genetically encoded indicators.
  JOURNAL   eLife. 2017 Jul 27;6. pii: e25690. doi: 10.7554/eLife.25690.
  PUBMED    28749338
REFERENCE   2  (bases 1 to 5843)
  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: "eLife."; 
            date: "2017-07-27"; volume: "6. pii: e25690. doi"; pages: " 
            10.7554/eLife.25690"
COMMENT     Sequence Label: pAAV-hSyn-ASAP2s
FEATURES             Location/Qualifiers
     source          1..5843
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        186..633
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      665..674
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1970..2558
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2023..2043)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2441..2452)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2590..3066
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2789..2808)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3106..3246
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3106..3235
                     /label=AAV2 ITR
     rep_origin      3321..3776
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3408..3427)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3618..3639
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3793..3812)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3912..3934
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3972..3990)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4058..4162
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4163..4231,4232..5023)
                     /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(4381..4400)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5194..5782
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5683..5702
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtgtgtc tagactgcag agggccctgc
      181 gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg
      241 accgaccccg acccactgga caagcaccca acccccattc cccaaattgc gcatccccta
      301 tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca
      361 ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa
      421 ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc
      481 gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac
      541 gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc
      601 agcggaggag tcgtgtcgtg cctgagagcg cagtcgagaa ggtaccggat cttctagagt
      661 cgacgccacc atggagacga ctgtgaggta tgaacagggg tcagagctca ctaaaacttc
      721 gagctctcca acagcagatg agcccacgat aaagattgat gatggtcgtg atgagggtaa
      781 tgaacaagac agctgttcca ataccattag gagaaaaatt tccccgtttg tgatgtcatt
      841 tggattcaga gtatttggag ttgtgcttat cattgtagac atcatagtgg tgattgtgga
      901 tctggccatc agtgagaaga aaagaggcat tagagagatt cttgaaggtg tttccctggc
      961 tatagcactc ttcttccttg ttgatgttct catgagagtg tttgttgaag gcttcaagaa
     1021 ctatttccgg tccaaactga atactttgga tgcagtcata gtagtgggca ctctgctaat
     1081 taatatgacc tactccttct ctgaccttgc tgcctttaac agccataacg tgtatattac
     1141 cgcggataaa cagaaaaacg gcattaaagc gaactttacc gtgcgccata acgtggaaga
     1201 tggcagcgtg cagctggcgg atcattatca gcagaacacc ccgattggcg atggcccggt
     1261 gctgctgccg gataaccatt atctgagcac ccagaccgtg ctgagcaaag atccgaacga
     1321 aaaacgcgat cacatggtgc tgctggaatt tgtgaccgca gcgggcatta cacacggcat
     1381 ggatgaactg tatggcggca ccggcggcag cgcgagccag ggcgaagaac tgtttaccgg
     1441 cgtggtgccg attctggtgg aactggatgg cgatgtgaac ggccataaat ttagcgtgcg
     1501 cggcgaaggc gaaggcgatg cgaccattgg caaactgacc ctgaaattta tttgcaccac
     1561 cggcaaacta ccggtgccgt ggccgaccct ggtgaccacc ttaacctatg gcgtgcagtg
     1621 ctttagccgc tatccggatc atatgaaacg ccatgatttt tttaaaagcg cgatgccgga
     1681 aggctatgtg caggaacgca ccattagctt taaagatgat ggcaaatata aaacccgcgc
     1741 ggtggtgaaa tttgaaggcg ataccctggt gaaccgcatt gaactgaaag gcaccgattt
     1801 taaagaagat ggcaacattc tggggcataa actggaatat aacacagatc agatgccgca
     1861 gatggttact cttcttcgag ttctgagaat tgttatctta ataagaatat ttcagctggc
     1921 ttcacagaag aaacaacttg aagtggtaac ctaatgaaag cttatcgata atcaacctct
     1981 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     2041 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2101 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     2161 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     2221 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     2281 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     2341 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     2401 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     2461 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     2521 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag cgctgctcga
     2581 gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt
     2641 gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac
     2701 taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt
     2761 gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca
     2821 caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct
     2881 cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg
     2941 tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc
     3001 tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct
     3061 gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga
     3121 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     3181 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     3241 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     3301 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3361 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3421 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3481 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     3541 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3601 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3661 gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3721 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     3781 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     3841 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     3901 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     3961 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     4021 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4081 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4141 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4201 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4261 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4321 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4381 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4441 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     4501 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4561 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     4621 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4681 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     4741 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     4801 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     4861 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     4921 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     4981 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     5041 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5101 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5161 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5221 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5281 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5341 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5401 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5461 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5521 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5581 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5641 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5701 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5761 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5821 ttgctggcct tttgctcaca tgt
//
