LOCUS       sequence-197043-        5833 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 5833)
  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 5833)
  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..5833
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        174..621
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      654..663
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1959..2547
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2012..2032)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2430..2441)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2579..3055
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2778..2797)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3095..3235
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3095..3224
                     /label=AAV2 ITR
     rep_origin      3310..3765
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3397..3416)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3607..3628
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3782..3801)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3901..3923
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3961..3979)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4047..4151
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4152..4220,4221..5012)
                     /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(4370..4389)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5183..5771
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5672..5691
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(5833,1..129)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc cgggcgtcgg gcgacctttg
       61 gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta
      121 ggggttcctg cggccgcacg cgtgtgtcta gactgcagag ggccctgcgt atgagtgcaa
      181 gtgggtttta ggaccaggat gaggcggggt gggggtgcct acctgacgac cgaccccgac
      241 ccactggaca agcacccaac ccccattccc caaattgcgc atcccctatc agagaggggg
      301 aggggaaaca ggatgcggcg aggcgcgtgc gcactgccag cttcagcacc gcggacagtg
      361 ccttcgcccc cgcctggcgg cgcgcgccac cgccgcctca gcactgaagg cgcgctgacg
      421 tcactcgccg gtcccccgca aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc
      481 gccggcccag ccggaccgca ccacgcgagg cgcgagatag gggggcacgg gcgcgaccat
      541 ctgcgctgcg gcgccggcga ctcagcgctg cctcagtctg cggtgggcag cggaggagtc
      601 gtgtcgtgcc tgagagcgca gtcgagaagg taccggatct gcatgagcta gccgccacca
      661 tggagacgac tgtgaggtat gaacaggggt cagagctcac taaaacttcg agctctccaa
      721 cagcagatga gcccacgata aagattgatg atggtcgtga tgagggtaat gaacaagaca
      781 gctgttccaa taccattagg agaaaaattt ccccgtttgt gatgtcattt ggattcagag
      841 tatttggagt tgtgcttatc attgtagaca tcatagtggt gattgtggat ctggccatca
      901 gtgagaagaa aagaggcatt agagagattc ttgaaggtgt ttccctggct atagcactct
      961 tcttccttgt tgatgttctc atgagagtgt ttgttgaagg cttcaagaac tatttccggt
     1021 ccaaactgaa tactttggat gcagtcatag tagtgggcac tctgctaatt aatatgacct
     1081 actccttctc tgaccttgct gcctttaaca gccataacgt gtatattacc gcggataaac
     1141 agaaaaacgg cattaaagcg aactttaccg tgcgccataa cgtggaagat ggcagcgtgc
     1201 agctggcgga tcattatcag cagaacaccc cgattggcga tggcccggtg ctgctgccgg
     1261 ataaccatta tctgagcacc cagaccgtgc tgagcaaaga tccgaacgaa aaacgcgatc
     1321 acatggtgct gctggaattt gtgaccgcag cgggcattac acacggcatg gatgaactgt
     1381 atggcggcac cggcggcagc gcgagccagg gcgaagaact gtttaccggc gtggtgccga
     1441 ttctggtgga actggatggc gatgtgaacg gccataaatt tagcgtgcgc ggcgaaggcg
     1501 aaggcgatgc gaccattggc aaactgaccc tgaaatttat ttgcaccacc ggcaaactac
     1561 cggtgccgtg gccgaccctg gtgaccacct taacctatgg cgtgcagtgc tttagccgct
     1621 atccggatca tatgaaacgc catgattttt ttaaaagcgc gatgccggaa ggctatgtgc
     1681 aggaacgcac cattagcttt aaagatgatg gcaaatataa aacccgcgcg gtggtgaaat
     1741 ttgaaggcga taccctggtg aaccgcattg aactgaaagg caccgatttt aaagaagatg
     1801 gcaacattct ggggcataaa ctggaatata acacagatca gatgccgcag atggttactc
     1861 ttcttcgagt tctgagaatt gttatcttaa taagaatatt tcagctggct tcacagaaga
     1921 aacaacttga agtggtaacc taatgaaagc ttatcgataa tcaacctctg gattacaaaa
     1981 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     2041 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     2101 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     2161 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     2221 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     2281 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     2341 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc acctggattc
     2401 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     2461 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     2521 ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag agatctacgg
     2581 gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt
     2641 gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt
     2701 ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac
     2761 ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct
     2821 cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt
     2881 gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg
     2941 tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg
     3001 gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat
     3061 tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc
     3121 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     3181 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     3241 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     3301 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3361 gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct
     3421 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3481 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     3541 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3601 ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt
     3661 ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac
     3721 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     3781 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     3841 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     3901 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     3961 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     4021 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4081 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4141 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4201 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4261 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4321 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4381 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4441 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4501 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4561 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4621 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4681 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4741 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4801 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4861 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4921 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4981 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5041 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5101 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5161 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5221 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5281 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5341 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5401 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5461 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5521 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5581 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5641 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5701 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5761 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5821 ttgctcacat gtc
//
