LOCUS       Exported                5835 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  in vivo voltage imaging.
ACCESSION   .
VERSION     .
KEYWORDS    DRH323: pAAV_hSyn-QuasAr2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5835)
  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 5835)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5835
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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)"
     CDS             645..1403
                     /codon_start=1
                     /product="achaerhodopsin-3 (Arch) derivative with improved 
                     voltage sensitivity (Hochbaum et al., 2014)"
                     /label=QuasAr2
                     /translation="MVSIALQAGYDLLGDGRPETLWLGIGTLLMLIGTFYFLVRGWGVT
                     DKDAREYYAVTILVSGIASAAYLSMFFGIGLTEVSVGGEMLDIYYARYAQWLFTTPLLL
                     LHLALLAKVDRVTIGTLVGVDALMIVTGLIGALSHTAIARYSWWLFSTICMIVVLYVLA
                     TSLRSAAKERGPEVASTFNTLTALVLVLWTAYPILWIIGTEGAGVVGLGIETLLFMVLD
                     VTAKVGFGFILLRSRAILGDTEAPEPSAGAD"
     CDS             1410..2117
                     /codon_start=1
                     /product="photostable monomeric orange derivative of DsRed 
                     fluorescent protein (Shaner et al., 2008)"
                     /label=mOrange2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEYNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPHFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GKIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     misc_feature    2226..2814
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2279..2299)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2697..2708)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2856..2873)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2862..3069
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3098..3238
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3098..3227
                     /label=AAV2 ITR
     rep_origin      3313..3768
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3400..3419)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3610..3631
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3785..3804)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3904..3926
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3964..3982)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4050..4154
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4155..5015
                     /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(4373..4392)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5186..5774
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5675..5694
                     /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 caccatggta agtatcgctc
      661 tgcaggctgg ttacgaccta ctgggtgacg gcagacctga aaccctgtgg ctgggcatcg
      721 gcactctgct gatgctgatt ggaaccttct actttctggt ccgcggatgg ggagtcaccg
      781 ataaggatgc ccgggaatat tacgctgtga ctatcctggt gtccggaatc gcatccgccg
      841 catatctgtc tatgttcttt ggtatcgggc ttactgaggt gtccgtcggg ggcgaaatgt
      901 tggatatcta ttatgccagg tacgcccagt ggctgtttac caccccactt ctgctgctgc
      961 acctggccct tctcgctaag gtggatcggg tgaccatcgg caccctggtg ggtgtggacg
     1021 ccctgatgat cgtcactggc ctcatcggag ccttgagcca cacggccata gccagataca
     1081 gttggtggtt gttctctaca atttgcatga tagtggtgct ctatgttctg gctacatccc
     1141 tgcgatctgc tgcaaaggag cggggccccg aggtggcatc tacctttaac accctgacag
     1201 ctctggtctt ggtgctgtgg accgcttacc ctatcctgtg gatcataggc actgagggcg
     1261 ctggcgtggt gggcctgggc atcgaaactc tgctgtttat ggtgttggac gtgactgcca
     1321 aggtcggctt tggctttatc ctgttgagat cccgggctat tctgggcgac accgaggcac
     1381 cagaacccag tgccggtgcc gatacgttga tggtgagcaa gggcgaggag tataacatgg
     1441 ccatcatcaa ggagttcatg cgcttcaagg tgcgcatgga gggctccgtg aacggccacg
     1501 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggctttcag accgctaagc
     1561 tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc cctcatttca
     1621 cctacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac ttcaagctgt
     1681 ccttccccga gggcttcaag tgggagcgcg tgatgaacta cgaggacggc ggcgtggtga
     1741 ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg
     1801 gcaccaactt cccctccgac ggccccgtga tgcagaagaa gaccatgggc tgggaggcct
     1861 cctccgagcg gatgtacccc gaggacggtg ccctgaaggg caagatcaag atgaggctga
     1921 agctgaagga cggcggccac tacacctccg aggtcaagac cacctacaag gccaagaagc
     1981 ccgtgcagct gcccggcgcc tacatcgtcg acatcaagtt ggacatcacc tcccacaacg
     2041 aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca
     2101 tggacgagct gtacaagagt agaatcacaa gcgaaggcga gtacatcccc ctggatcaaa
     2161 tagacataaa tgtaggtgga ttttgttatg agaatgaata agaattcgat atcaagctta
     2221 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     2281 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     2341 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     2401 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     2461 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     2521 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     2581 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     2641 tgctcgccta tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     2701 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     2761 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     2821 accgagcgct gctcgagcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca
     2881 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     2941 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     3001 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     3061 tgctggggag gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg
     3121 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     3181 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     3241 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     3301 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3361 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     3421 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     3481 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     3541 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3601 tctttaatag tggactcttg ttccaaactg gaacaacact caatcctatc tcgggctatt
     3661 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     3721 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     3781 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     3841 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     3901 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     3961 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     4021 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4081 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4141 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4201 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4261 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4321 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4381 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4441 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     4501 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4561 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     4621 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4681 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     4741 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4801 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4861 gtgagcgtgg aagccgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     4921 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4981 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     5041 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5101 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5161 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5221 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5281 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     5341 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5401 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5461 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5521 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5581 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5641 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5701 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5761 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5821 cttttgctca catgt
//
