LOCUS       sequence-209180-        6501 bp    DNA     linear   UNA 15-DEC-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 6501)
  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 6501)
  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-hSynapsin- soCoChR-GFP
FEATURES             Location/Qualifiers
     source          1..6501
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     polyA_signal    71..278
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     267..284
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     misc_feature    317..905
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             423..434
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     832..852
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(942..1652,1653..1655))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             complement(join(942..1652,1653..1655))
                     /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(984..1005)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1333..1352
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1592..1613
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(1674..2123)
                     /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(2160..3023)
                     /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(3057..3504)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     misc_feature    3540..3631
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(3572..3591)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    3645..3693
                     /note="synthetic polyadenylation signal"
     repeat_region   3749..3878
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3940..4528)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4020..4039)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4699..5490,5491..5559))
                     /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     5322..5341
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5560..5664)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5732..5750
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5788..5810)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5910..5929
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5996..6451)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6133..6154)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6345..6364
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cctcttcgct attacgccag gctgcagaga gggagtggcc aactccatca ctaggggttc
       61 ctcagatctc tccccagcat gcctgctatt gtcttcccaa tcctccccct tgctgtcctg
      121 ccccacccca ccccccagaa tagaatgaca cctactcaga caatgcgatg caatttcctc
      181 attttattag gaaaggacag tgggagtggc accttccagg gtcaaggaag gcacggggga
      241 ggggcaaaca acagatggct ggcaactaga aggcacagtc gaggctgatc agcgagctct
      301 agtcgacggt atcgatgcgg ggaggcggcc caaagggaga tccgactcgt ctgagggcga
      361 aggcgaagac gcggaagagg ccgcagagcc ggcagcaggc cgcgggaagg aaggtccgct
      421 ggattgaggg ccgaagggac gtagcagaag ggcgtcccgc gcagaatcca ggtggcaaca
      481 caggcgagca gccaaggaaa ggacgatgat ttccccgaca acaccacgga attgtcagtg
      541 cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag ttccgccgtg
      601 gcaataggga gggggaaagc gaaagtcccg gaaaggagct gacaggtggt ggcaatgccc
      661 caaccagtgg gggttgcgtc agcaaacaca gtgcacacca cgccacgttg cctgacaacg
      721 ggccacaact cctcataaag agacagcaac caggatttat acaaggagga gaaaatgaaa
      781 gccatacggg aagcaatagc atgatacaaa ggcattaaag cagcgtatcc acatagcgta
      841 aaaggagcaa catagttaag aataccagtc aatctttcac aaattttgta atccagaggt
      901 tgattatcga taagctttta cacttcattc tcgtagcaga acttgtacag ctcgtccatg
      961 ccgagagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg
     1021 ttggggtctt tgctcagggc ggactgggtg ctcaggtagt ggttgtcggg cagcagcacg
     1081 gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc
     1141 tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt tcttctgctt gtcggccatg
     1201 atatagacgt tgtggctgtt gtagttgtac tccagcttgt gccccaggat gttgccgtcc
     1261 tccttgaagt cgatgccctt cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc
     1321 acctcggcgc gggtcttgta gttgccgtcg tccttgaaga agatggtgcg ctcctggacg
     1381 tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg
     1441 ctgaagcact gcacgccgta ggtcagggtg gtcacgaggg tgggccaggg cacgggcagc
     1501 ttgccggtgg tgcaaatgaa cttcagggtc agcttgccgt aggtggcatc gccctcgccc
     1561 tcgccggaca cgctgaactt gtggccgttt acgtcgccgt ccagctcgac caggatgggc
     1621 accaccccgg tgaacagctc ctcgcccttg ctcacagtac ctccgctagc tccgctgact
     1681 gccagtgaca catcctcgtt tgatggatac aggctcactg aggcgaagcg caggtactgc
     1741 agcctgggtg tttcctctgg cccgaccttg atatggggga tttccttctc gccacaaatg
     1801 tgagacacgg tacttgcaga agcaggagag gagcttggtc ccaggacgga gaccactcct
     1861 tttggcagga tctggcacat ggtgtctgtg gtctcgtact ggctatctct ctgcagttcg
     1921 aaaatgtcca cctcgacccg ggcctttgca ggcacttcaa tgatgccatt gatctgctcc
     1981 ctagccagtg ccagtgccag cctctctcct cttccgcaca cggtctgatc gtccagaata
     2041 gcggccatcc tcagagaaga cagcacctgg caagaagggt tagcaaaagc gacaatcagc
     2101 agcagcagca gttcggcggg catggtacct ccacttccac cggttccacc tgaacctccc
     2161 actgtctcct cgtcctcctg gtccaccatg gtctccactt ccatctcctc gccggccacg
     2221 ttgatcttgg tcttcttccg gatgtcgccg tagatgatga tgtgctggtg gatcagcact
     2281 ctcagatagt gtcccagcag gccccaacaa ttcttgctca tgaggtcgat gatggtgtgg
     2341 ccaatggtag agccgtacac gctgatggcg tcgaatcctt ctggtcccac gacgaacagc
     2401 acggggaaca tgccccaaga caggaagaac agccaagcca tgatccgcac gacggttcta
     2461 ggtctgccct ttggcaccac gtggtagctc tcgatataca ccttggcggc gtggaagaag
     2521 gtgttggcgc cgtagatgca acccagcacg aagaagatga ccttgacgta gcctgtgctc
     2581 atggcgcttg tagctcccca cacgatggtt cccacgtctg acaccagcag cctcatggtc
     2641 cgcttgctgt agtcgtcctt caggccggtc aggttgctca ggtggatcag gatgacggga
     2701 caagtcagca gccactctgc gtatctcagc cactggactc ggtgtccgtt agccaggtac
     2761 agcatgctgg ggtcgtcgaa ctcgtggaag aactcgatga tgaccttggt cagctcgacg
     2821 caacagacgt agacctcctc ccaaccgcaa gtggctctcc aagtctggta ggcgtagaac
     2881 atcaggatca ggatgctgaa gccgaaggcg aaccactgga ggatgttggc caccagctgc
     2941 tctgtatcgc ttcccaggct gtcggtccaa gccaggcaaa agcactggtc gataggcaca
     3001 atggcgctgc cgttccccag catggtggcg gatccttgct agcagcttga attcgactgc
     3061 gctctcaggc acgacacgac tcctccgctg cccaccgcag actgaggcag cgctgagtcg
     3121 ccggcgccgc agcgcagatg gtcgcgcccg tgccccccta tctcgcgcct cgcgtggtgc
     3181 ggtccggctg ggccggcggc ggcgcggacg cgaccaaggt ggccgggaag gggagtttgc
     3241 gggggaccgg cgagtgacgt cagcgcgcct tcagtgctga ggcggcggtg gcgcgcgccg
     3301 ccaggcgggg gcgaaggcac tgtccgcggt gctgaagctg gcagtgcgca cgcgcctcgc
     3361 cgcatcctgt ttcccctccc cctctctgat aggggatgcg caatttgggg aatgggggtt
     3421 gggtgcttgt ccagtgggtc ggggtcggtc gtcaggtagg cacccccacc ccgcctcatc
     3481 ctggtcctaa aacccacttg cactcatacg cagggccctc tgcagctcga gtctagaata
     3541 gagaaatgtt ctggcacctg cacttgcact ggggacagcc tattttgcta gtttgttttg
     3601 tttcgttttg ttttgatgga gagcgtatgt tagtactatc gattcacaca aaaaaccaac
     3661 acacagatgt aatgaaaata aagatatttt attgcggccg ctccaagtac ctcccgtacc
     3721 ttaatatttc tagaacgcgt gcggccgcag gaacccctag tgatggagtt ggccactccc
     3781 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     3841 ggcctcagtg agcgagcgag cgcgcagctg cctgcaggac atgtgagcaa aaggccagca
     3901 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3961 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4021 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4081 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4141 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4201 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4261 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4321 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4381 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4441 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4501 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4561 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4621 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4681 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4741 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4801 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4861 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4921 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4981 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5041 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5101 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5161 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5221 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5281 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5341 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5401 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5461 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5521 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5581 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5641 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     5701 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     5761 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     5821 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     5881 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     5941 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt
     6001 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     6061 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     6121 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     6181 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     6241 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     6301 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     6361 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     6421 gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg cgcaactgtt
     6481 gggaagggcg atcggtgcgg g
//
