LOCUS       sequence-361691-        7850 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Voltage indicator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7850)
  AUTHORS   Fan LZ, Kim DK, Jennings JH, Tian H, Wang PY, Ramakrishnan C, 
            Randles S, Sun Y, Thadhani E, Kim YS, Quirin S, Giocomo L, Cohen AE,
            Deisseroth K
  TITLE     All-optical physiology resolves a synaptic basis for behavioral 
            timescale plasticity.
  JOURNAL   Cell. 2023 Feb 2;186(3):543-559.e19. doi: 
            10.1016/j.cell.2022.12.035. Epub 2023 Jan 19.
  PUBMED    36669484
REFERENCE   2  (bases 1 to 7850)
  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"; doi: 
            "10.1016/j.cell.2022.12.035"; journalName: "Cell"; date: "2023-02-2-
            2"; volume: "186"; issue: "3"; pages: "543-559.e19"
COMMENT     Sequence Label: pAAV_hSyn-fDIO-somQuasAr6a-EGFP-P2A-sombC1C2TG
FEATURES             Location/Qualifiers
     source          1..7850
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    654..687
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             complement(978..1004)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(1932..1988)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(join(2238..2948,2949..2951))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             complement(join(2238..2948,2949..2951))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2280..2301)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2629..2648
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2888..2909
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3815..3824
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3835..3868)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    3977..4565
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4030..4050)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4448..4459)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4597..5073
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4796..4815)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5113..5253
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5113..5242
                     /label=AAV2 ITR
     rep_origin      5328..5783
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5415..5434)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5625..5646
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5800..5819)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5919..5941
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5979..5997)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6065..6169
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6170..6238,6239..7030)
                     /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(6388..6407)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7201..7789
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7690..7709
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gatcctctag agtcgactcc ggagaagttc
      661 ctattctcta gaaagtatag gaacttcgca gaatggtagc tggattgtag ctgctattag
      721 caatatgaaa cctcttagaa gttcctattc ttcaaaaggt ataggaactt cggcgcgcct
      781 tagaatcgag tagcctctgg aaaatcggtg gcgcatgaaa tgaagctgtc gatactggac
      841 atagatcgca tgtctataac gccttcagtc cgtgctggaa ggggtgcgac agggctgggc
      901 atggatgaca tctccaattc ctcgggtggt tttgattgtg gggccatttc cttagtgtta
      961 agtataggct gactctgagc gtaatctgga acatcgtatg ggtagaccgc gccagcctcg
     1021 gcctcgtcct ccaccagcgt ctcgacctca atctcagtgc caccaatgtt caatttggtg
     1081 gtcttgcgaa tgtcgccgtg gatgaggata tgctcgtgga tcagcacgcg caggtagtgg
     1141 ccgagcagac cccagcagtt cttcgacatc aggtcaatga tggtgtggcc gacggtggag
     1201 ccgtacacgc tcaggacgcc gaagccctcg gggccgagga tgaacaggat ggggaacata
     1261 ccccatgata cgaagaagag ccaagccatg ccagtcacca cctggcgaca ccggcccttc
     1321 ggcacggtgt ggtacgcctc aatgtagacc ttggctgcgt tgaagaatgt gtagatgccg
     1381 tagcacaggc ccatcaggaa gaaaatgaca cggacgtatc ccttggacag cgcggccgtg
     1441 gtgccccaca cgatgccgcc gatatctgac accagcagac ccatggtacg cttgttatag
     1501 tcgttcgcca gacccgtaag gttgctcaga tggataagaa cgacagggca ggtcagcagc
     1561 cactccgcgt aacgaagcca cacggtcttg ttgccgttgg atgagtagat caccgcaggt
     1621 tcgtcaaact catggaaata ctcgatgatg aacttgatca tctcgatcgt ggccacgtaa
     1681 atctcctccc agccgcaagt agacttccag gtctggtagc cgtagaacat caggcagagc
     1741 gctgaaagcg caaaagtaat ccactgcaga atgttggcag ccaacttctc ggcatttgtt
     1801 ccgttggatt taagccaagc caagcagaag cactggccgt tgttcgggat gcaaataaca
     1861 gaaccattgt tctcaagagt gtatgaggtt tggaagagtc cgacggcaga caaagcgccg
     1921 ccatagtcca tgggaccggg gttttcttcc acgtctcctg cttgctttaa cagagagaag
     1981 ttcgtggctc cggagcccac ctcgttctcg tagcagaact tgtagaacct ggtggcctca
     2041 gggaagtcgg tggcgcagct gatgaagctg tcgatgctgc tcatgctcct catgtcgatc
     2101 acgccctcgg tcctggcagg cagaggggcc acagggctag gcatgctgct catctccagc
     2161 tcctcaggag gcttgctctg aggggccatc tccttggtgt tcaggatagg ctggctctgg
     2221 gtacctccgc tcccaccctt gtacagctcg tccatgccga gagtgatccc ggcggcggtc
     2281 acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct cagggcggac
     2341 tgggtgctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc
     2401 tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag
     2461 ttcaccttga tgccgttctt ctgcttgtcg gccatgatat agacgttgtg gctgttgtag
     2521 ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc
     2581 tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg
     2641 ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg
     2701 aagaagtcgt gctgcttcat gtggtcgggg tagcggctga agcactgcac gccgtaggtc
     2761 agggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca aatgaacttc
     2821 agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg
     2881 ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg
     2941 cccttgctca ccacgttgat gtcgatctgg tccaggggga tgtactcgcc ctcgctggtg
     3001 atcctgctct tggcggccgc cttgctcact gctactaccg ggcggtcggc ggcaccggta
     3061 tcagctccag ctgaaggttc aggtgcttct gtatcttgca gaatgcttct tgacctgagc
     3121 aggataaaac cgaagcccac cttggcggtg acgtccagca ccatgaacag caaagtttcg
     3181 attcccaggc ccacaactcc ggcaccctcg gtgccgataa tccagatgat agggtaagca
     3241 gtccacagga ccagaaccag tgcggtgagg atgttgaagg ttgaggcaac ctctgggcct
     3301 ctctccttag cggctgaccg cagagaagtg gccaggacat acagaacgac gatcatacag
     3361 atagttgaga acaggcacca agagtatctg gcaatggcag tatgtgacag tgtccctatc
     3421 aggcctgtga caatcatcag agcatcgccg ccaacgagag tgccgatggt cactctatca
     3481 actttggcca ggagtgccag gtgcagcagc aacagtggtg tagtcagcag ccactgtgcg
     3541 tatcttgcgt agtagatgtt caggatctct ccacctactg gcacctctgt gaggccaata
     3601 ccaaagaaca ttgacagata agctgcactg gcaataccgc tgaccaggat tgtcactgcg
     3661 tagtactccc gtgcatcctt gtcagtctct ccgcccgctc taaccaggaa gtagaaggtt
     3721 ccgatcagca tcagcagtgt accaattccg agccacagac tttcaggtct accatctccg
     3781 aggaggtcat aaccagcctg cagtgcgata gaaaccatgg tggcggaggc tagcgaagtt
     3841 cctatacttt ctagagaata ggaacttctt gccttaaccc agaaattatc actgttattc
     3901 tttagaatgg tgcaaagaga agttcctata ccttttgaag aataggaact tcgaattcga
     3961 tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     4021 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     4081 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     4141 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     4201 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     4261 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     4321 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     4381 ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt ccttctgcta
     4441 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     4501 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     4561 cccgcatcga taccgagcgc tgctcgagag atctacgggt ggcatccctg tgacccctcc
     4621 ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata
     4681 aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg
     4741 ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat
     4801 tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg
     4861 gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc
     4921 aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg
     4981 gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta
     5041 caggcgtgaa ccactgctcc cttccctgtc cttctgattt tgtaggtaac cacgtgcgga
     5101 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     5161 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     5221 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     5281 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     5341 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     5401 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     5461 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     5521 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     5581 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     5641 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
     5701 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     5761 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     5821 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     5881 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     5941 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     6001 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     6061 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     6121 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     6181 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     6241 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     6301 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     6361 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     6421 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     6481 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     6541 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     6601 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     6661 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     6721 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     6781 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     6841 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     6901 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     6961 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     7021 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     7081 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     7141 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     7201 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     7261 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     7321 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     7381 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     7441 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     7501 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     7561 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     7621 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     7681 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     7741 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     7801 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
