LOCUS       sequence-219838-        6559 bp    DNA     linear   UNA 14-APR-2025
DEFINITION  A somatic channelrhodopsin, which enables single cell, single 
            millisecond resolution optogenetics. Human Synapsin (hSynapsin) 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6559)
  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 6559)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-hSynapsin- soCoChR-GFP
FEATURES             Location/Qualifiers
     source          1..6559
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     polyA_signal    109..316
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     305..322
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     misc_feature    355..943
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             461..472
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     870..890
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(980..1690,1691..1693))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             complement(join(980..1690,1691..1693))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1022..1043)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1371..1390
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1630..1651
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(1712..2161)
                     /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             complement(2198..3061)
                     /codon_start=1
                     /product="high-photocurrent channelrhodopsin from 
                     Chloromonas oogama (Shemesh et al., 2017)"
                     /label=CoChR
                     /translation="MLGNGSAIVPIDQCFCLAWTDSLGSDTEQLVANILQWFAFGFSIL
                     ILMFYAYQTWRATCGWEEVYVCCVELTKVIIEFFHEFDDPSMLYLANGHRVQWLRYAEW
                     LLTCPVILIHLSNLTGLKDDYSKRTMRLLVSDVGTIVWGATSAMSTGYVKVIFFVLGCI
                     YGANTFFHAAKVYIESYHVVPKGRPRTVVRIMAWLFFLSWGMFPVLFVVGPEGFDAISV
                     YGSTIGHTIIDLMSKNCWGLLGHYLRVLIHQHIIIYGDIRKKTKINVAGEEMEVETMVD
                     QEDEETV"
     promoter        complement(3095..3542)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     misc_feature    3578..3669
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(3610..3629)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    3683..3731
                     /note="synthetic polyadenylation signal"
     repeat_region   3787..3916
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3978..4566)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4058..4077)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4737..5528,5529..5597))
                     /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     5360..5379
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5598..5702)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5770..5788
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5826..5848)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5948..5967
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6034..6489)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6171..6192)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6383..6402
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg
       61 cgcagagagg gagtggccaa ctccatcact aggggttcct cagatctctc cccagcatgc
      121 ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc ccccagaata
      181 gaatgacacc tactcagaca atgcgatgca atttcctcat tttattagga aaggacagtg
      241 ggagtggcac cttccagggt caaggaaggc acgggggagg ggcaaacaac agatggctgg
      301 caactagaag gcacagtcga ggctgatcag cgagctctag tcgacggtat cgatgcgggg
      361 aggcggccca aagggagatc cgactcgtct gagggcgaag gcgaagacgc ggaagaggcc
      421 gcagagccgg cagcaggccg cgggaaggaa ggtccgctgg attgagggcc gaagggacgt
      481 agcagaaggg cgtcccgcgc agaatccagg tggcaacaca ggcgagcagc caaggaaagg
      541 acgatgattt ccccgacaac accacggaat tgtcagtgcc caacagccga gcccctgtcc
      601 agcagcgggc aaggcaggcg gcgatgagtt ccgccgtggc aatagggagg gggaaagcga
      661 aagtcccgga aaggagctga caggtggtgg caatgcccca accagtgggg gttgcgtcag
      721 caaacacagt gcacaccacg ccacgttgcc tgacaacggg ccacaactcc tcataaagag
      781 acagcaacca ggatttatac aaggaggaga aaatgaaagc catacgggaa gcaatagcat
      841 gatacaaagg cattaaagca gcgtatccac atagcgtaaa aggagcaaca tagttaagaa
      901 taccagtcaa tctttcacaa attttgtaat ccagaggttg attatcgata agcttttaca
      961 cttcattctc gtagcagaac ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg
     1021 tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg
     1081 actgggtgct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt
     1141 tctgctggta gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg cggatcttga
     1201 agttcacctt gatgccgttc ttctgcttgt cggccatgat atagacgttg tggctgttgt
     1261 agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca
     1321 gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt
     1381 tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact
     1441 tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc acgccgtagg
     1501 tcagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg caaatgaact
     1561 tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt
     1621 ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct
     1681 cgcccttgct cacagtacct ccgctagctc cgctgactgc cagtgacaca tcctcgtttg
     1741 atggatacag gctcactgag gcgaagcgca ggtactgcag cctgggtgtt tcctctggcc
     1801 cgaccttgat atgggggatt tccttctcgc cacaaatgtg agacacggta cttgcagaag
     1861 caggagagga gcttggtccc aggacggaga ccactccttt tggcaggatc tggcacatgg
     1921 tgtctgtggt ctcgtactgg ctatctctct gcagttcgaa aatgtccacc tcgacccggg
     1981 cctttgcagg cacttcaatg atgccattga tctgctccct agccagtgcc agtgccagcc
     2041 tctctcctct tccgcacacg gtctgatcgt ccagaatagc ggccatcctc agagaagaca
     2101 gcacctggca agaagggtta gcaaaagcga caatcagcag cagcagcagt tcggcgggca
     2161 tggtacctcc acttccaccg gttccacctg aacctcccac tgtctcctcg tcctcctggt
     2221 ccaccatggt ctccacttcc atctcctcgc cggccacgtt gatcttggtc ttcttccgga
     2281 tgtcgccgta gatgatgatg tgctggtgga tcagcactct cagatagtgt cccagcaggc
     2341 cccaacaatt cttgctcatg aggtcgatga tggtgtggcc aatggtagag ccgtacacgc
     2401 tgatggcgtc gaatccttct ggtcccacga cgaacagcac ggggaacatg ccccaagaca
     2461 ggaagaacag ccaagccatg atccgcacga cggttctagg tctgcccttt ggcaccacgt
     2521 ggtagctctc gatatacacc ttggcggcgt ggaagaaggt gttggcgccg tagatgcaac
     2581 ccagcacgaa gaagatgacc ttgacgtagc ctgtgctcat ggcgcttgta gctccccaca
     2641 cgatggttcc cacgtctgac accagcagcc tcatggtccg cttgctgtag tcgtccttca
     2701 ggccggtcag gttgctcagg tggatcagga tgacgggaca agtcagcagc cactctgcgt
     2761 atctcagcca ctggactcgg tgtccgttag ccaggtacag catgctgggg tcgtcgaact
     2821 cgtggaagaa ctcgatgatg accttggtca gctcgacgca acagacgtag acctcctccc
     2881 aaccgcaagt ggctctccaa gtctggtagg cgtagaacat caggatcagg atgctgaagc
     2941 cgaaggcgaa ccactggagg atgttggcca ccagctgctc tgtatcgctt cccaggctgt
     3001 cggtccaagc caggcaaaag cactggtcga taggcacaat ggcgctgccg ttccccagca
     3061 tggtggcgga tccttgctag cagcttgaat tcgactgcgc tctcaggcac gacacgactc
     3121 ctccgctgcc caccgcagac tgaggcagcg ctgagtcgcc ggcgccgcag cgcagatggt
     3181 cgcgcccgtg cccccctatc tcgcgcctcg cgtggtgcgg tccggctggg ccggcggcgg
     3241 cgcggacgcg accaaggtgg ccgggaaggg gagtttgcgg gggaccggcg agtgacgtca
     3301 gcgcgccttc agtgctgagg cggcggtggc gcgcgccgcc aggcgggggc gaaggcactg
     3361 tccgcggtgc tgaagctggc agtgcgcacg cgcctcgccg catcctgttt cccctccccc
     3421 tctctgatag gggatgcgca atttggggaa tgggggttgg gtgcttgtcc agtgggtcgg
     3481 ggtcggtcgt caggtaggca cccccacccc gcctcatcct ggtcctaaaa cccacttgca
     3541 ctcatacgca gggccctctg cagctcgagt ctagaataga gaaatgttct ggcacctgca
     3601 cttgcactgg ggacagccta ttttgctagt ttgttttgtt tcgttttgtt ttgatggaga
     3661 gcgtatgtta gtactatcga ttcacacaaa aaaccaacac acagatgtaa tgaaaataaa
     3721 gatattttat tgcggccgct ccaagtacct cccgtacctt aatatttcta gaacgcgtgc
     3781 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     3841 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggcgg cctcagtgag cgagcgagcg
     3901 cgcagctgcc tgcaggacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3961 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4021 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4081 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4141 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4201 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4261 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4321 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4381 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     4441 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4501 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4561 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4621 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4681 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     4741 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4801 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     4861 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     4921 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     4981 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5041 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5101 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5161 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5221 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5281 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     5341 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     5401 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     5461 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     5521 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     5581 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     5641 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     5701 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     5761 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     5821 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     5881 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     5941 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     6001 cacagatgcg taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa
     6061 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     6121 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     6181 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     6241 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     6301 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     6361 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     6421 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     6481 agggcgcgtc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     6541 ctcttcgcta ttacgccag
//
