LOCUS       Exported                7411 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  in vivo voltage imaging.
ACCESSION   .
VERSION     .
KEYWORDS    YA1627: pAAV_hSyn-Dio-paQuasAr3-s-P2A-CheRiff-s
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7411)
  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 7411)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7411
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        186..633
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    671..704
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    755..788
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(995..1021)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(1037..1981)
                     /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             complement(1982..2038)
                     /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(2264..2977)
                     /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(2306..2327)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2655..2674
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2914..2935
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(2984..3742)
                     /codon_start=1
                     /product="achaerhodopsin-3 (Arch) derivative with improved 
                     voltage sensitivity (Hochbaum et al., 2014)"
                     /label=QuasAr2
                     /translation="MVSIALQAGYDLLGDGRPETLWLGIGTLLMLIGTFYFLVRGWGVT
                     DKDAREYYAVTILASGIASAAYLSMFFGIGLTEVSVGGEMLDIYYARYAQWLFTTPLLL
                     LHLALLAKVDRVTIGTLVGVDALMIVTGLIGALSHTAIARYSWWLFSTICMIVVLYVLA
                     TSLRSAAKERGPEVASTFNTLTALVLVLWTAYPILWIIGTEGAGVVGLGIETLLFMVLD
                     VTAKVGFGFILLRSRAILGDTEAPEPSAGAD"
     regulatory      3739..3748
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    complement(3758..3791)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    complement(3842..3875)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3900..4488
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3953..3973)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4371..4382)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4542..4561)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4596..4615
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4674..4814
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4674..4803
                     /label=AAV2 ITR
     rep_origin      4889..5344
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4976..4995)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5186..5207
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5361..5380)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5480..5502
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5540..5558)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5626..5730
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5731..6591
                     /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(5949..5968)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6762..7350
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7251..7270
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtgtgtc tagactgcag agggccctgc
      181 gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg
      241 accgaccccg acccactgga caagcaccca acccccattc cccaaattgc gcatccccta
      301 tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca
      361 ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa
      421 ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc
      481 gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac
      541 gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc
      601 agcggaggag tcgtgtcgtg cctgagagcg cagtcgagaa ggtaccggat cctctagagt
      661 cgactccgga ataacttcgt ataggatact ttatacgaag ttatgcagaa tggtagctgg
      721 attgtagctg ctattagcaa tatgaaacct cttaataact tcgtatagca tacattatac
      781 gaagttatgg cgcgccttag aatcgagtag cctctggaaa atcggtggcg catgaaatga
      841 agctgtcgat actggacata gatcgcatgt ctataacgcc ttcagtccgt gctggaaggg
      901 gtgcgacagg gctgggcatg gatgacatct ccaattcctc gggtggtttt gattgtgggg
      961 ccatttcctt agtgttaagt ataggctgac tctgagcgta atctggaaca tcgtatgggt
     1021 agctcttggc ggccgccact ttatcttcct ctgtcacgaa ggccagcacg tccaccttgc
     1081 gtcccagaaa ctgggagctc actggtctcc tgatatcgcc gtacataatg atatgctcgt
     1141 gaattttgat ccgcaggaag tgccccagca gtccccaaat attcttggac agcaggtcgg
     1201 cgatggtatg gccaatagtg gacccagcca cgctcagggt gtgcatgcct tcaggcccaa
     1261 agatgaacag gccgggaaac atcagccaac tagaatagta cacggcagtc atggccagca
     1321 ccagtttctt acagccgccg gcaggcatga tatagtaaga ctccacgtag atgattccag
     1381 cgttgtagaa tgtctgggtt ccatacacca ggccgataca gtaaaacagc cacttcaccc
     1441 acccagtggc cagagcggct gtcactccca tgcaaatagt tcccaggtcg gacaccagca
     1501 gagccattgt ccgcttattg tatgcctcac tcaggccggt gatgttagac agatgaatca
     1561 ggatcacggg acatgtcagc agccattcgg catatctcag ccatggggta atgttccctc
     1621 cgtacaggaa cagcatagca ggactggtga actcgaaata gatctccaga atcactttga
     1681 tcagctccac ggagcacacg tacacttcct cccatcccac gctggcctta taggcgtgcc
     1741 agccgtagaa agccagaaac agggcagaca ggaacacggc aaaccactgg aagcccctag
     1801 ctgtttcctg ctcccacagg gccccatagt agatgtagat ggagctgcac tgttcctcat
     1861 acccatgcac ggggtggtag ggtggattca cttgatatcc agctggggca tggtactcac
     1921 tgcctccggc agaatcgttt ccaggtgggg cgctgtgagc gtctggagca ggagctccgc
     1981 cgggaccggg gttttcttcc acgtctcctg cttgctttaa cagagagaag ttcgtggctc
     2041 cggagcccac ttcattctca taacaaaaga atctggtggc ttcagggaag tccgtggcac
     2101 agctgatgaa gctgtcaatg ctggacatgc tccgcatgtc gatgacgccc tccgtgcgtg
     2161 cgggcagagg ggccacgggg ctgggcatgc tgctcatctc cagctcctct ggaggcttgc
     2221 tctgcggggc catctccttg gtgttgagga tgggctggga ctgcttgtac agctcgtcca
     2281 tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct
     2341 cgttggggtc tttgctcagg gcggactggt agctcaggta gtggttgtcg ggcagcagca
     2401 cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt
     2461 cctcgatgtt gtggcggatc ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca
     2521 tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt
     2581 cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
     2641 tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga
     2701 cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc
     2761 gggcgaagca catcaggccg tagccgaagg tggtcacgag ggtgggccag ggcacgggca
     2821 gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc
     2881 cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg
     2941 gcaccacccc ggtgaacagc tcctcgccct tgctcaccgg aggatcggca ccggcactgg
     3001 gttctggtgc ctcggtgtcg cccagaatag cccgggatct caacaggata aagccaaagc
     3061 cgaccttggc agtcacgtcc aacaccataa acagcagagt ttcgatgccc aggcccacca
     3121 cgccagcgcc ctcagtgcct atgatccaca ggatagggta agcggtccac agcaccaaga
     3181 ccagagctgt cagggtgtta aaggtagatg ccacctcggg gccccgctcc tttgcagcag
     3241 atcgcaggga tgtagccaga acatagagca ccactatcat gcaaattgta gagaacaacc
     3301 accaactgta tctggctatg gccgtgtggc tcaaggctcc gatgaggcca gtgacgatca
     3361 tcagggcgtc cacacccacc agggtgccga tggtcacccg atccacctta gcgagaaggg
     3421 ccaggtgcag cagcagaagt ggggtggtaa acagccactg ggcgtacctg gcataataga
     3481 tatccaacat ttcgcccccg acggacacct cagtaagccc gataccaaag aacatagaca
     3541 gatatgcggc ggatgcgatt ccggaggcca ggatagtcac agcgtaatat tcccgggcat
     3601 ccttatcggt gactccccat ccgcggacca gaaagtagaa ggttccaatc agcatcagca
     3661 gagtgccgat gcccagccac agggtttcag gtctgccgtc acccagtagg tcgtaaccag
     3721 cctgcagagc gatacttacc atggtggcgg agctagcata acttcgtata aagtatccta
     3781 tacgaagtta tttgccttaa cccagaaatt atcactgtta ttctttagaa tggtgcaaag
     3841 aataacttcg tataatgtat gctatacgaa gttatgaatt cgatatcaag cttatcgata
     3901 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     3961 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     4021 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     4081 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     4141 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     4201 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     4261 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     4321 cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     4381 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     4441 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc
     4501 gacccgggcg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     4561 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     4621 ttgtaaccat tataagctgc aataaactaa ccacgtgcgg accgagcggc cgcaggaacc
     4681 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     4741 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     4801 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4861 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     4921 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     4981 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     5041 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     5101 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     5161 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     5221 cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     5281 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     5341 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     5401 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     5461 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     5521 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     5581 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5641 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5701 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5761 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5821 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5881 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5941 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     6001 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     6061 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     6121 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     6181 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     6241 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     6301 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     6361 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6421 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6481 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6541 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6601 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6661 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6721 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6781 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6841 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6901 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6961 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     7021 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     7081 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     7141 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     7201 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     7261 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     7321 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     7381 ctggcctttt gctggccttt tgctcacatg t
//
