LOCUS       sequence-222463-        6538 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  in vivo voltage imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6538)
  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 6538)
  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: YA1531: pAAV_CAG-FLEX-QuasAr3
FEATURES             Location/Qualifiers
     source          1..6538
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     279..301
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     363..382
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    417..450
                     /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    513..546
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(799..1509,1510..1512))
                     /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(841..862)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1190..1209
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1449..1470
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(1582..2340)
                     /codon_start=1
                     /product="achaerhodopsin-3 (Arch) derivative with improved 
                     voltage sensitivity (Hochbaum et al., 2014)"
                     /label=QuasAr2
                     /translation="MVSIALQAGYDLLGDGRPETLWLGIGTLLMLIGTFYFLVRGWGVT
                     DKDAREYYAVTILVSGIASAAYLSMFFGIGLTEVSVGGEMLDIYYARYAQWLFTTPLLL
                     LHLALLAKVDRVTIGTLVGVDALMIVTGLIGALSHTAIARYSWWLFSTICMIVVLYVLA
                     TSLRSAARERGPEVASTFNTLTALVLVLWTAYPILWIIGTEGAGVVGLGIETLLFMVLD
                     VTAKVGFGFILLRSRAILGDTEAPEPSAGAD"
     regulatory      2337..2346
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2363..2396)
                     /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(2467..2500)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2542..3130
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2595..2615)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3013..3024)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3173..3294
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3184..3203)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3238..3257
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3456..3585
                     /label=AAV2 ITR
     rep_origin      complement(3822..4410)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3902..3921)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4581..5372,5373..5441))
                     /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     5204..5223
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5442..5546)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5572..6027)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5709..5730)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5921..5940
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6099..6228
                     /label=AAV2 ITR
ORIGIN
        1 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
       61 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
      121 ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa
      181 agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg gctgtccgcg
      241 gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac
      301 cggcggctct agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct
      361 gggcaacgtg ctggttattg tgctgtctca tcattttggc aaagaattgg atccccataa
      421 cttcgtataa agtatcctat acgaagttat atcaaaatag gaagaccaat gcttcaccat
      481 cgacccgaat tgccaagcat caccatcgac ccataacttc gtataatgta tgctatacga
      541 agttatacta gggatccccc ctcgagtttt acttgtacat tacacttcat tctcataaca
      601 aaagcccccc acattgatgt caatctgatc gaggggtata tactcgccct cagaagtgat
      661 tcgacttttt ccccccacgt ttatatcgat ttgatccaga gggatgtact cgccctcaga
      721 ggttatgcgg gattttccac cgacgttaat gtcgatctga tccagaggga tatactcgcc
      781 ctcagaggtg atccttgatt tgtacagctc gtccatgccg agagtgatcc cggcggcggt
      841 cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga
      901 ctggtagctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt
      961 ctgctggtag tggtcggcga gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa
     1021 gttcaccttg atgccgttct tctgcttgtc ggccatgata tagacgttgt ggctgttgta
     1081 gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag
     1141 ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt
     1201 gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt
     1261 gaagaagtcg tgctgcttca tgtggtcggg gtagcgggcg aagcacatca ggccgtagcc
     1321 gaaggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt
     1381 cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg
     1441 gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc
     1501 gcccttgctc actacattta tgtctatttg atccaggggg atgtactcgc cttcgcttgt
     1561 gattctactc ttcatcaacg tatcggcacc ggcactgggt tctggtgcct cggtgtcgcc
     1621 cagaatagcc cgggatctca acaggataaa gccaaagccg accttggcag tcacgtccaa
     1681 caccataaac agcagagttt cgatgcccag gcccaccacg ccagcgccct cagtgcctat
     1741 gatccacagg atagggtaag cggtccacag caccaagacc agagctgtca gggtgttaaa
     1801 ggtagatgcc acctcggggc cccgctccct tgcagcagat cgcagggatg tagccagaac
     1861 atagagcacc actatcatgc aaattgtaga gaacaaccac caactgtatc tggctatggc
     1921 cgtgtggctc aaggctccga tgaggccagt gacgatcatc agggcgtcca cacccaccag
     1981 ggtgccgatg gtcacccgat ccaccttagc gagaagggcc aggtgcagca gcagaagtgg
     2041 ggtggtaaac agccactggg cgtacctggc ataatagata tccaacattt cgcccccgac
     2101 ggacacctca gtaagcccga taccaaagaa catagacaga tatgcggcgg atgcgattcc
     2161 ggacaccagg atagtcacag cgtaatattc ccgggcatcc ttatcggtga ctccccatcc
     2221 gcggaccaga aagtagaagg ttccaatcag catcagcaga gtgccgatgc ccagccacag
     2281 ggtttcaggt ctgccgtcac ccagtaggtc gtaaccagcc tgcagagcga tacttaccat
     2341 ggtggcggta ccgaattcta gtataacttc gtataggata ctttatacga agttatcatt
     2401 gggattcttc ctattttgat ccaagcatca ccatcgaccc tctagtccag atctcaccat
     2461 cgacccataa cttcgtatag catacattat acgaagttat gtccctcgaa gaggttcgaa
     2521 ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
     2581 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     2641 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     2701 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     2761 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     2821 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     2881 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     2941 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     3001 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     3061 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     3121 gcctccccgc atcgataccg tcgacccggg cggccgcttc gagcagacat gataagatac
     3181 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     3241 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     3301 aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc
     3361 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc ttcctagagc atggctacgt
     3421 agataagtag catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc
     3481 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     3541 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgca ttaatgaatc
     3601 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     3661 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3721 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3781 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3841 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3901 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3961 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4021 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4081 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4141 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4201 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4261 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4321 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4381 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4441 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4501 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4561 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4621 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4681 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4741 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4801 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4861 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4921 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4981 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5041 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5101 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5161 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5221 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5281 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5341 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5401 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5461 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5521 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
     5581 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     5641 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     5701 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     5761 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     5821 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     5881 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     5941 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     6001 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg
     6061 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct gcgcgctcgc tcgctcactg
     6121 aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg
     6181 agcgagcgcg cagagaggga gtggccaact ccatcactag gggttccttg tagttaatga
     6241 ttaacccgcc atgctactta tctacgtagc catgctctag gaagagtacc attgacgtca
     6301 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     6361 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     6421 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     6481 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatgg
//
