LOCUS       sequence-386149-        6040 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetically-encoded voltage indicator SpikeyGi2, soma and AIS 
            targeted, double-flowed in AAV vector under the promoter hSyn.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6040)
  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 6040)
  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_Flex_syn_SpikeyGi2_kv2.1
FEATURES             Location/Qualifiers
     source          1..6040
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(263..851)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(343..362)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1022..1813,1814..1882))
                     /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     1645..1664
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1883..1987)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2013..2468)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2150..2171)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2362..2381
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2540..2669
                     /label=AAV2 ITR
     promoter        2772..3219
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    3235..3268
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    3331..3364
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      4894..4903
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(4910..4943)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(5014..5047)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5080..5668
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5133..5153)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5551..5562)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5709..5916
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 atggctacgt agataagtag catggcgggt taatcattaa ctacaaggaa cccctagtga
       61 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgactcgctg cgctcggtcg
      121 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
      181 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
      241 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
      301 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
      361 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
      421 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
      481 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
      541 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
      601 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
      661 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
      721 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
      781 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
      841 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
      901 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
      961 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     1021 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     1081 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     1141 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     1201 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     1261 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     1321 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     1381 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     1441 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     1501 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     1561 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     1621 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     1681 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     1741 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     1801 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     1861 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     1921 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     1981 ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa
     2041 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     2101 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     2161 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     2221 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     2281 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     2341 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     2401 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     2461 gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     2521 ctcttcgcta ttacgccagc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc
     2581 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
     2641 agtggccaac tccatcacta ggggttcctt gtagttaatg attaacccgc catgctactt
     2701 atctacgtag ccatgctcta ggaagatcgt aggttaatta agtgtctaga ctgcagaggg
     2761 ccctgcgtat gagtgcaagt gggttttagg accaggatga ggcggggtgg gggtgcctac
     2821 ctgacgaccg accccgaccc actggacaag cacccaaccc ccattcccca aattgcgcat
     2881 cccctatcag agagggggag gggaaacagg atgcggcgag gcgcgtgcgc actgccagct
     2941 tcagcaccgc ggacagtgcc ttcgcccccg cctggcggcg cgcgccaccg ccgcctcagc
     3001 actgaaggcg cgctgacgtc actcgccggt cccccgcaaa ctccccttcc cggccacctt
     3061 ggtcgcgtcc gcgccgccgc cggcccagcc ggaccgcacc acgcgaggcg cgagataggg
     3121 gggcacgggc gcgaccatct gcgctgcggc gccggcgact cagcgctgcc tcagtctgcg
     3181 gtgggcagcg gaggagtcgt gtcgtgcctg agagcgcagt cgagaagaat tcccataact
     3241 tcgtataaag tatcctatac gaagttatat caaaatagga agaccaatgc ttcaccatcg
     3301 acccgaattg ccaagcatca ccatcgaccc ataacttcgt ataatgtatg ctatacgaag
     3361 ttatactagc tagttatcaa aagcgggtcg cttccggaaa atcggtcgcg cagctaataa
     3421 agctatcaat gctgctcatg ctgcgcatat caatcacgcc ttcggtgcgc gccggcagcg
     3481 gcgccaccgg gctcggcatg ctgctcattt ccagttcttc cggcggtttg ctctgcggcg
     3541 ccatttcttt ggtgttcaga atcggctggc tctgacgcgt ggaagatcca gatgaacctg
     3601 aagaaccctt aagggttacc acttcaagtt gtttcttctg tgaagccagc tgaaatattc
     3661 ttattaagat aacaattctc agaactcgaa gaagagtaac catctgcggc atctgatctg
     3721 tgttatattc cagtttatgc cccagaatgt tgccatcttc tttaaaatcg gtgcctttca
     3781 gttcaatgcg gttcaccagg gtatcgcctt caaatttcac caccgcgcgg gttttatatt
     3841 tgccatcatc tttaaagcta atggtgcgtt cctgcacata gccttccggc atcgcgcttt
     3901 taaaaaaatc atggcgtttc atatgatccg gatagcggct aaagcactgc acgccatagg
     3961 ttaaggtggt caccagggtc ggccacggca ccggtagttt gccggtggtg caaataaatt
     4021 tcagggtcag tttgccaatg gtcgcatcgc cttcgccttc gccgcgcacg ctaaatttat
     4081 ggccgttcac atcgccatcc agttccacca gaatcggcac cacgccggta aacagttctt
     4141 cgccctggct cgcgctgccg ccggtgccgc catacagttc atccatgccg tgtgtaatgc
     4201 ccgctgcggt cacaaattcc agcagcacca tgtgatcgcg tttttcgttc ggatctttgc
     4261 tcagcacggt ctgggtgctc agataatggt tatccggcag cagcaccggg ccatcgccaa
     4321 tcggggtgtt ctgctgataa tgatccgcca gctgcacgct gccatcttcc acgttatggc
     4381 gaatggtaaa gttcgcttta atgccgtttt tctgtttatc cgcggtaata tacacgttac
     4441 tatcatcacc gcgaaatgta ccgtcagaga aggagtaggt catattaatt agcagagtgc
     4501 ccactactat gactgcatcc aaagtattca gtttggaccg gaaatagttc ttgaagcctt
     4561 caacaaacac tctcatgaga acatcaacaa ggaagaagag tgctatagcc agggaaacac
     4621 cttcaagaat ctctctaatg cctcttttct tctcactgat ggccagatcc acaatcacca
     4681 ctatgatgtc tacaatgata agcacaactc caaatactct gaatccaaat gacatcacaa
     4741 acggggaaat ttttctccta atggtattgg aacagctgtc ttgttcatta ccctcatcac
     4801 gaccatcatc aatctttatc gtgggctcat ctgctgttgg agagctcgaa gttttagtga
     4861 gctctgaccc ctgttcatac ctcacagtcg tctccatggt ggcgctagta taacttcgta
     4921 taggatactt tatacgaagt tatcattggg attcttccta ttttgatcca agcatcacca
     4981 tcgaccctct agtccagatc tcaccatcga cccataactt cgtatagcat acattatacg
     5041 aagttatgtc cctcgaagag gttcgcgaag cttatcgata atcaacctct ggattacaaa
     5101 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     5161 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     5221 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     5281 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     5341 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     5401 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     5461 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
     5521 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     5581 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     5641 cggatctccc tttgggccgc ctccccgcat cgataccgtc gacccgggcg gccgctcata
     5701 aagctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     5761 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     5821 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     5881 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtccggactg tacaattgca
     5941 ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc
     6001 tctacaaatg tggtaaaatc gataaggatc ttcctagagc
//
