LOCUS       sequence-209415-        6465 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  A somatic channelrhodopsin, which enables single cell, single 
            millisecond resolution optogenetics. hSynapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6465)
  AUTHORS   Shemesh OA, Tanese D, Zampini V, Linghu C, Piatkevich K, Ronzitti E,
            Papagiakoumou E, Boyden ES, Emiliani V
  TITLE     Temporally precise single-cell-resolution optogenetics.
  JOURNAL   Nat Neurosci. 2017 Dec;20(12):1796-1806. doi: 
            10.1038/s41593-017-0018-8. Epub 2017 Nov 13.
  PUBMED    29184208
REFERENCE   2  (bases 1 to 6465)
  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: 
            "10.1038/s41593-017-0018-8"; journalName: "Nat Neurosci"; date: 
            "2017-12"; volume: "20"; issue: "12"; pages: "1796-1806"
COMMENT     Sequence Label: pAAV-hSyn-soCoChR-mCardinal
FEATURES             Location/Qualifiers
     source          1..6465
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     rep_origin      30..485
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(117..136)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     327..348
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(552..571)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     671..693
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(731..749)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        817..921
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(922..990,991..1782)
                     /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(1140..1159)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1953..2541
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2442..2461
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2603..2732
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     polyA_signal    2788..2836
                     /note="synthetic polyadenylation signal"
     misc_feature    2850..2941
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2890..2909
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        2977..3424
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             3458..4321
                     /codon_start=1
                     /product="high-photocurrent channelrhodopsin from 
                     Chloromonas oogama (Shemesh et al., 2017)"
                     /label=CoChR
                     /translation="MLGNGSAIVPIDQCFCLAWTDSLGSDTEQLVANILQWFAFGFSIL
                     ILMFYAYQTWRATCGWEEVYVCCVELTKVIIEFFHEFDDPSMLYLANGHRVQWLRYAEW
                     LLTCPVILIHLSNLTGLKDDYSKRTMRLLVSDVGTIVWGATSAMSTGYVKVIFFVLGCI
                     YGANTFFHAAKVYIESYHVVPKGRPRTVVRIMAWLFFLSWGMFPVLFVVGPEGFDAISV
                     YGSTIGHTIIDLMSKNCWGLLGHYLRVLIHQHIIIYGDIRKKTKINVAGEEMEVETMVD
                     QEDEETV"
     CDS             4358..4807
                     /codon_start=1
                     /product="first 150 residues of the mouse kainate receptor 
                     subunit 2"
                     /label=KA2
                     /note="mediates targeting to the neuronal soma (Shemesh et 
                     al., 2017)"
                     /translation="MPAELLLLLIVAFANPSCQVLSSLRMAAILDDQTVCGRGERLALA
                     LAREQINGIIEVPAKARVEVDIFELQRDSQYETTDTMCQILPKGVVSVLGPSSSPASAS
                     TVSHICGEKEIPHIKVGPEETPRLQYLRFASVSLYPSNEDVSLAVS"
     CDS             4826..5557
                     /codon_start=1
                     /product="far-red fluorescent protein derived from 
                     mNeptune, with further red-shifted spectra"
                     /label=mCardinal
                     /translation="VSKGEELIKENMHMKLYMEGTVNNHHFKCTTEGEGKPYEGTQTQR
                     IKVVEGGPLPFAFDILATCFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGV
                     LTVTQDTSLQDGCLIYNVKLRGVNFPSNGPVMQKKTLGWEATTETLYPADGGLEGRCDM
                     ALKLVGGGHLHCNLKTTYRSKKPAKNLKMPGVYFVDRRLERIKEADNETYVEQHEVAVA
                     RYCDLPSKLGHKLNGMDELYK"
     misc_feature    5570..6158
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5623..5643)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6041..6052)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6191..6208)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6197..6404
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 aacagttgcg cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc
       61 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
      121 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
      181 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
      241 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
      301 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
      361 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
      421 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
      481 aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
      541 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
      601 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
      661 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
      721 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
      781 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
      841 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
      901 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
      961 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     1021 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     1081 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     1141 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     1201 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     1261 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     1321 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     1381 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     1441 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     1501 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     1561 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     1621 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     1681 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     1741 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     1801 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     1861 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     1921 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     1981 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     2041 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     2101 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2161 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2221 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2281 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     2341 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     2401 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     2461 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     2521 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     2581 tgctggcctt ttgctcacat gtcctgcagg cagctgcgcg ctcgctcgct cactgaggcc
     2641 gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga
     2701 gggagtggcc aactccatca ctaggggttc ctgcggccgc acgcgttcta gaaatattaa
     2761 ggtacgggag gtacttggag cggccgcaat aaaatatctt tattttcatt acatctgtgt
     2821 gttggttttt tgtgtgaatc gatagtacta acatacgctc tccatcaaaa caaaacgaaa
     2881 caaaacaaac tagcaaaata ggctgtcccc agtgcaagtg caggtgccag aacatttctc
     2941 tattctagac tcgagctgca gagggccctg cgtatgagtg caagtgggtt ttaggaccag
     3001 gatgaggcgg ggtgggggtg cctacctgac gaccgacccc gacccactgg acaagcaccc
     3061 aacccccatt ccccaaattg cgcatcccct atcagagagg gggaggggaa acaggatgcg
     3121 gcgaggcgcg tgcgcactgc cagcttcagc accgcggaca gtgccttcgc ccccgcctgg
     3181 cggcgcgcgc caccgccgcc tcagcactga aggcgcgctg acgtcactcg ccggtccccc
     3241 gcaaactccc cttcccggcc accttggtcg cgtccgcgcc gccgccggcc cagccggacc
     3301 gcaccacgcg aggcgcgaga taggggggca cgggcgcgac catctgcgct gcggcgccgg
     3361 cgactcagcg ctgcctcagt ctgcggtggg cagcggagga gtcgtgtcgt gcctgagagc
     3421 gcagtcgaat tcaagctgct agcaaggatc cgccaccatg ctggggaacg gcagcgccat
     3481 tgtgcctatc gaccagtgct tttgcctggc ttggaccgac agcctgggaa gcgatacaga
     3541 gcagctggtg gccaacatcc tccagtggtt cgccttcggc ttcagcatcc tgatcctgat
     3601 gttctacgcc taccagactt ggagagccac ttgcggttgg gaggaggtct acgtctgttg
     3661 cgtcgagctg accaaggtca tcatcgagtt cttccacgag ttcgacgacc ccagcatgct
     3721 gtacctggct aacggacacc gagtccagtg gctgagatac gcagagtggc tgctgacttg
     3781 tcccgtcatc ctgatccacc tgagcaacct gaccggcctg aaggacgact acagcaagcg
     3841 gaccatgagg ctgctggtgt cagacgtggg aaccatcgtg tggggagcta caagcgccat
     3901 gagcacaggc tacgtcaagg tcatcttctt cgtgctgggt tgcatctacg gcgccaacac
     3961 cttcttccac gccgccaagg tgtatatcga gagctaccac gtggtgccaa agggcagacc
     4021 tagaaccgtc gtgcggatca tggcttggct gttcttcctg tcttggggca tgttccccgt
     4081 gctgttcgtc gtgggaccag aaggattcga cgccatcagc gtgtacggct ctaccattgg
     4141 ccacaccatc atcgacctca tgagcaagaa ttgttggggc ctgctgggac actatctgag
     4201 agtgctgatc caccagcaca tcatcatcta cggcgacatc cggaagaaga ccaagatcaa
     4261 cgtggccggc gaggagatgg aagtggagac catggtggac caggaggacg aggagacagt
     4321 gggaggttca ggtggaaccg gtggaagtgg aggtaccatg cccgccgaac tgctgctgct
     4381 gctgattgtc gcttttgcta acccttcttg ccaggtgctg tcttctctga ggatggccgc
     4441 tattctggac gatcagaccg tgtgcggaag aggagagagg ctggcactgg cactggctag
     4501 ggagcagatc aatggcatca ttgaagtgcc tgcaaaggcc cgggtcgagg tggacatttt
     4561 cgaactgcag agagatagcc agtacgagac cacagacacc atgtgccaga tcctgccaaa
     4621 aggagtggtc tccgtcctgg gaccaagctc ctctcctgct tctgcaagta ccgtgtctca
     4681 catttgtggc gagaaggaaa tcccccatat caaggtcggg ccagaggaaa cacccaggct
     4741 gcagtacctg cgcttcgcct cagtgagcct gtatccatca aacgaggatg tgtcactggc
     4801 agtcagcgga gctagcggag gtactgtgag caagggcgag gagctgatca aggagaacat
     4861 gcacatgaag ctgtacatgg aaggcaccgt gaacaaccac cacttcaagt gcaccaccga
     4921 aggggagggc aagccctacg agggcaccca gacccagagg attaaggtgg tggagggagg
     4981 ccccctgccg ttcgcattcg acatcctggc cacctgcttt atgtacggga gcaagacctt
     5041 catcaaccac acccagggca tccccgattt ctttaagcag tccttccctg agggcttcac
     5101 atgggagaga gtcaccacat acgaagacgg gggcgtgctt accgttaccc aggacaccag
     5161 cctccaggac ggctgcttga tctacaacgt caagctcaga ggggtgaact tcccatccaa
     5221 cggccctgtg atgcagaaga aaacactcgg ctgggaggcc accaccgaga ccctgtaccc
     5281 cgctgacggc ggcctggaag gcagatgcga catggccctg aagctcgtgg gcgggggcca
     5341 cctgcactgc aacctgaaga ccacatacag atccaagaaa cccgctaaga acctcaagat
     5401 gcccggcgtc tactttgtgg accgcagact ggaaagaatc aaggaggccg acaatgagac
     5461 ctacgtcgag cagcacgagg tggctgtggc cagatactgc gacctcccta gcaaactggg
     5521 gcacaaactt aatggcatgg acgagctgta caagtaaaag cttatcgata atcaacctct
     5581 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     5641 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     5701 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     5761 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     5821 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     5881 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     5941 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     6001 cacctggatt ctgcgcggga cgcccttctg ctacgtccct tcggccctca atccagcgga
     6061 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     6121 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc gactagagct
     6181 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     6241 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     6301 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     6361 agcaaggggg aggattggga agacaatagc aggcatgctg gggagagatc tgaggaaccc
     6421 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctc
//
