LOCUS       sequence-217193-        7152 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  in vivo voltage imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7152)
  AUTHORS   Adam Y, Kim JJ, Lou S, Zhao Y, Xie ME, Brinks D, Wu H, Mostajo-Radji
            MA, Kheifets S, Parot V, Chettih S, Williams KJ, Gmeiner B, Farhi 
            SL, Madisen L, Buchanan EK, Kinsella I, Zhou D, Paninski L, Harvey 
            CD, Zeng H, Arlotta P, Campbell RE, Cohen AE
  TITLE     Voltage imaging and optogenetics reveal behaviour-dependent changes 
            in hippocampal dynamics.
  JOURNAL   Nature. 2019 May 1. pii: 10.1038/s41586-019-1166-7. doi: 
            10.1038/s41586-019-1166-7.
  PUBMED    31043747
REFERENCE   2  (bases 1 to 7152)
  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: "Nature. 
            2019 May 1. pii: 10.1038/s41586-019-1166-7. doi: 
            10.1038/s41586-019-1166-7."
COMMENT     Sequence Label: YA1529: pAAV_hSyn-QuasAr3-P2A-CheRiff
FEATURES             Location/Qualifiers
     source          1..7152
                     /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        181..628
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      642..651
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             648..1406
                     /codon_start=1
                     /product="achaerhodopsin-3 (Arch) derivative with improved 
                     voltage sensitivity (Hochbaum et al., 2014)"
                     /label=QuasAr2
                     /translation="MVSIALQAGYDLLGDGRPETLWLGIGTLLMLIGTFYFLVRGWGVT
                     DKDAREYYAVTILVSGIASAAYLSMFFGIGLTEVSVGGEMLDIYYARYAQWLFTTPLLL
                     LHLALLAKVDRVTIGTLVGVDALMIVTGLIGALSHTAIARYSWWLFSTICMIVVLYVLA
                     TSLRSAARERGPEVASTFNTLTALVLVLWTAYPILWIIGTEGAGVVGLGIETLLFMVLD
                     VTAKVGFGFILLRSRAILGDTEAPEPSAGAD"
     CDS             join(1476..1478,1479..2189)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1518..1539)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1779..1798)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2126..2147
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2415..2471
                     /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             2472..3416
                     /codon_start=1
                     /product="blue light-activated optogenetic actuator derived
                     from Scherffelia dubia channelrhodopsin (Hochbaum et al., 
                     2014)"
                     /label=CheRiff
                     /translation="GGAPAPDAHSAPPGNDSAGGSEYHAPAGYQVNPPYHPVHGYEEQC
                     SSIYIYYGALWEQETARGFQWFAVFLSALFLAFYGWHAYKASVGWEEVYVCSVELIKVI
                     LEIYFEFTSPAMLFLYGGNITPWLRYAEWLLTCPVILIHLSNITGLSEAYNKRTMALLV
                     SDLGTICMGVTAALATGWVKWLFYCIGLVYGTQTFYNAGIIYVESYYIMPAGGCKKLVL
                     AMTAVYYSSWLMFPGLFIFGPEGMHTLSVAGSTIGHTIADLLSKNIWGLLGHFLRIKIH
                     EHIIMYGDIRRPVSSQFLGRKVDVLAFVTEEDKV"
     CDS             3489..3515
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    3543..4131
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3596..3616)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4014..4025)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4173..4190)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4179..4386
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4415..4555
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4415..4544
                     /label=AAV2 ITR
     rep_origin      4630..5085
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4717..4736)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4927..4948
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5102..5121)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5221..5243
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5281..5299)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5367..5471
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5472..5540,5541..6332)
                     /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(5690..5709)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6503..7091
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6992..7011
                     /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 gcgttgtaca gtgtctagac tgcagagggc cctgcgtatg
      181 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
      241 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
      301 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg
      361 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
      421 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
      481 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
      541 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
      601 aggagtcgtg tcgtgcctga gagcgcagtc gagaaggatc cgccaccatg gtaagtatcg
      661 ctctgcaggc tggttacgac ctactgggtg acggcagacc tgaaaccctg tggctgggca
      721 tcggcactct gctgatgctg attggaacct tctactttct ggtccgcgga tggggagtca
      781 ccgataagga tgcccgggaa tattacgctg tgactatcct ggtgtccgga atcgcatccg
      841 ccgcatatct gtctatgttc tttggtatcg ggcttactga ggtgtccgtc gggggcgaaa
      901 tgttggatat ctattatgcc aggtacgccc agtggctgtt taccacccca cttctgctgc
      961 tgcacctggc ccttctcgct aaggtggatc gggtgaccat cggcaccctg gtgggtgtgg
     1021 acgccctgat gatcgtcact ggcctcatcg gagccttgag ccacacggcc atagccagat
     1081 acagttggtg gttgttctct acaatttgca tgatagtggt gctctatgtt ctggctacat
     1141 ccctgcgatc tgctgcaagg gagcggggcc ccgaggtggc atctaccttt aacaccctga
     1201 cagctctggt cttggtgctg tggaccgctt accctatcct gtggatcata ggcactgagg
     1261 gcgctggcgt ggtgggcctg ggcatcgaaa ctctgctgtt tatggtgttg gacgtgactg
     1321 ccaaggtcgg ctttggcttt atcctgttga gatcccgggc tattctgggc gacaccgagg
     1381 caccagaacc cagtgccggt gccgatacgt tgatgaagag tagaatcaca agcgaaggcg
     1441 agtacatccc cctggatcaa atagacataa atgtagtgag caagggcgag gagctgttca
     1501 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     1561 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     1621 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttcggc tacggcctga
     1681 tgtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1741 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     1801 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1861 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     1921 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     1981 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     2041 gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc gccctgagca
     2101 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     2161 tcactctcgg catggacgag ctgtacaaat caaggatcac ctctgagggc gagtatatcc
     2221 ctctggatca gatcgacatt aacgtcggtg gaaaatcccg cataacctct gagggcgagt
     2281 acatccctct ggatcaaatc gatataaacg tggggggaaa aagtcgaatc acttctgagg
     2341 gcgagtatat acccctcgat cagattgaca tcaatgtggg gggcttttgt tatgagaatg
     2401 aagtgggctc cggagccacg aacttctctc tgttaaagca agcaggagac gtggaagaaa
     2461 accccggtcc cggcggagct cctgctccag acgctcacag cgccccacct ggaaacgatt
     2521 ctgccggagg cagtgagtac catgccccag ctggatatca agtgaatcca ccctaccacc
     2581 ccgtgcatgg gtatgaggaa cagtgcagct ccatctacat ctactatggg gccctgtggg
     2641 agcaggaaac agctaggggc ttccagtggt ttgccgtgtt cctgtctgcc ctgtttctgg
     2701 ctttctacgg ctggcacgcc tataaggcca gcgtgggatg ggaggaagtg tacgtgtgct
     2761 ccgtggagct gatcaaagtg attctggaga tctatttcga gttcaccagt cctgctatgc
     2821 tgttcctgta cggagggaac attaccccat ggctgagata tgccgaatgg ctgctgacat
     2881 gtcccgtgat cctgattcat ctgtctaaca tcaccggcct gagtgaggca tacaataagc
     2941 ggacaatggc tctgctggtg tccgacctgg gaactatttg catgggagtg acagccgctc
     3001 tggccactgg gtgggtgaag tggctgtttt actgtatcgg cctggtgtat ggaacccaga
     3061 cattctacaa cgctggaatc atctacgtgg agtcttacta tatcatgcct gccggcggct
     3121 gtaagaaact ggtgctggcc atgactgccg tgtactattc tagttggctg atgtttcccg
     3181 gcctgttcat ctttgggcct gaaggcatgc acaccctgag cgtggctggg tccactattg
     3241 gccataccat cgccgacctg ctgtccaaga atatttgggg actgctgggg cacttcctgc
     3301 ggatcaaaat tcacgagcat atcattatgt acggcgatat caggagacca gtgagctccc
     3361 agtttctggg acgcaaggtg gacgtgctgg ccttcgtgac agaggaagat aaagtggcgg
     3421 ccgccaagag caggatcacc agcgagggcg agtacatccc cctggaccag atcgacatca
     3481 acgtggggta cccatacgat gttccagatt acgcttaaga attcgatatc aagcttatcg
     3541 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     3601 ctccttttac gctatgtgga tacgctgctt taatgacttt gtatcatgct attgcttccc
     3661 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     3721 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     3781 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     3841 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     3901 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     3961 tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     4021 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     4081 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc
     4141 gagcgctgct cgagctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
     4201 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     4261 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     4321 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     4381 tggggaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc
     4441 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     4501 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     4561 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     4621 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4681 gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct
     4741 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4801 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     4861 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4921 ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt
     4981 ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac
     5041 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     5101 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     5161 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     5221 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     5281 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     5341 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     5401 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     5461 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5521 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5581 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5641 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     5701 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5761 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5821 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5881 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5941 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     6001 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     6061 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     6121 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     6181 agcgtggaag ccgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     6241 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     6301 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     6361 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     6421 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6481 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6541 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6601 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6661 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6721 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6781 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6841 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6901 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6961 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     7021 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     7081 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     7141 ttgctcacat gt
//
