LOCUS       sequence-386148-        6056 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetically-encoded voltage indicator SpikeyGi2, soma and AIS 
            targeted, in AAV vector under the neuronal-specific promoter hSyn.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6056)
  AUTHORS   Platisa J, Ye X, Ahrens AM, Liu C, Chen IA, Davison IG, Tian L, 
            Pieribone VA, Chen JL
  TITLE     High-speed low-light in vivo two-photon voltage imaging of large 
            neuronal populations.
  JOURNAL   Nat Methods. 2023 Mar 27. doi: 10.1038/s41592-023-01820-3.
  PUBMED    36973547
REFERENCE   2  (bases 1 to 6056)
  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: "Nat 
            Methods. 2023 Mar 27. doi: 10.1038/s41592-023-01820-3."
COMMENT     Sequence Label: pAAV_hSyn_SpikeyGi2_kv2.1
FEATURES             Location/Qualifiers
     source          1..6056
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   14..143
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        188..635
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      655..664
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2196..2784
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2249..2269)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2667..2678)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2816..3292
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3015..3034)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3332..3472
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3332..3461
                     /label=AAV2 ITR
     rep_origin      3547..4002
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3634..3653)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3844..3865
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4019..4038)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4138..4160
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4198..4216)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4284..4388
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4389..4457,4458..5249)
                     /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(4607..4626)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5420..6008
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5909..5928
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttgctcaca tgtcctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg
       61 tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc
      121 caactccatc actaggggtt cctgcggccg cacgcgtgtg tctagactgc agagggccct
      181 gcgtatgagt gcaagtgggt tttaggacca ggatgaggcg gggtgggggt gcctacctga
      241 cgaccgaccc cgacccactg gacaagcacc caacccccat tccccaaatt gcgcatcccc
      301 tatcagagag ggggagggga aacaggatgc ggcgaggcgc gtgcgcactg ccagcttcag
      361 caccgcggac agtgccttcg cccccgcctg gcggcgcgcg ccaccgccgc ctcagcactg
      421 aaggcgcgct gacgtcactc gccggtcccc cgcaaactcc ccttcccggc caccttggtc
      481 gcgtccgcgc cgccgccggc ccagccggac cgcaccacgc gaggcgcgag ataggggggc
      541 acgggcgcga ccatctgcgc tgcggcgccg gcgactcagc gctgcctcag tctgcggtgg
      601 gcagcggagg agtcgtgtcg tgcctgagag cgcagtcgag aaggtaccgg atccgccacc
      661 atggagacga ctgtgaggta tgaacagggg tcagagctca ctaaaacttc gagctctcca
      721 acagcagatg agcccacgat aaagattgat gatggtcgtg atgagggtaa tgaacaagac
      781 agctgttcca ataccattag gagaaaaatt tccccgtttg tgatgtcatt tggattcaga
      841 gtatttggag ttgtgcttat cattgtagac atcatagtgg tgattgtgga tctggccatc
      901 agtgagaaga aaagaggcat tagagagatt cttgaaggtg tttccctggc tatagcactc
      961 ttcttccttg ttgatgttct catgagagtg tttgttgaag gcttcaagaa ctatttccgg
     1021 tccaaactga atactttgga tgcagtcata gtagtgggca ctctgctaat taatatgacc
     1081 tactccttct ctgacggtac atttcgcggt gatgatagta acgtgtatat taccgcggat
     1141 aaacagaaaa acggcattaa agcgaacttt accattcgcc ataacgtgga agatggcagc
     1201 gtgcagctgg cggatcatta tcagcagaac accccgattg gcgatggccc ggtgctgctg
     1261 ccggataacc attatctgag cacccagacc gtgctgagca aagatccgaa cgaaaaacgc
     1321 gatcacatgg tgctgctgga atttgtgacc gcagcgggca ttacacacgg catggatgaa
     1381 ctgtatggcg gcaccggcgg cagcgcgagc cagggcgaag aactgtttac cggcgtggtg
     1441 ccgattctgg tggaactgga tggcgatgtg aacggccata aatttagcgt gcgcggcgaa
     1501 ggcgaaggcg atgcgaccat tggcaaactg accctgaaat ttatttgcac caccggcaaa
     1561 ctaccggtgc cgtggccgac cctggtgacc accttaacct atggcgtgca gtgctttagc
     1621 cgctatccgg atcatatgaa acgccatgat ttttttaaaa gcgcgatgcc ggaaggctat
     1681 gtgcaggaac gcaccattag ctttaaagat gatggcaaat ataaaacccg cgcggtggtg
     1741 aaatttgaag gcgataccct ggtgaaccgc attgaactga aaggcaccga ttttaaagaa
     1801 gatggcaaca ttctggggca taaactggaa tataacacag atcagatgcc gcagatggtt
     1861 actcttcttc gagttctgag aattgttatc ttaataagaa tatttcagct ggcttcacag
     1921 aagaaacaac ttgaagtggt aacccttaag ggttcttcag gttcatctgg atcttccacg
     1981 cgtcagagcc agccgattct gaacaccaaa gaaatggcgc cgcagagcaa accgccggaa
     2041 gaactggaaa tgagcagcat gccgagcccg gtggcgccgc tgccggcgcg caccgaaggc
     2101 gtgattgata tgcgcagcat gagcagcatt gatagcttta ttagctgcgc gaccgatttt
     2161 ccggaagcga cccgcttttg ataaagctta tcgataatca acctctggat tacaaaattt
     2221 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     2281 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     2341 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     2401 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     2461 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     2521 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     2581 tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc tggattctgc
     2641 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     2701 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     2761 tctccctttg ggccgcctcc ccgcatcgat accgagcgct gctcgagaga tctacgggtg
     2821 gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc
     2881 caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta
     2941 taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg
     3001 tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac
     3061 tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg
     3121 attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt
     3181 caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc
     3241 tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt
     3301 gtaggtaacc acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact
     3361 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     3421 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
     3481 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     3541 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     3601 tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc
     3661 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3721 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     3781 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3841 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg
     3901 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
     3961 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     4021 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     4081 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     4141 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     4201 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     4261 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4321 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4381 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4441 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4501 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4561 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4621 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4681 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     4741 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     4801 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     4861 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     4921 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     4981 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     5041 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     5101 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     5161 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     5221 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     5281 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5341 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5401 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5461 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5521 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5581 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5641 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5701 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5761 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5821 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5881 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5941 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6001 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggcct
//
