LOCUS       sequence-258722-        6607 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A somatic channelrhodopsin, which enables single cell, single 
            millisecond resolution optogenetics. Human Synapsin (hSynapsin) 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6607)
  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 6607)
  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..6607
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    175..223
                     /note="synthetic polyadenylation signal"
     misc_feature    237..328
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     277..296
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        364..811
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             845..1708
                     /codon_start=1
                     /product="high-photocurrent channelrhodopsin from 
                     Chloromonas oogama (Shemesh et al., 2017)"
                     /label=CoChR
                     /translation="MLGNGSAIVPIDQCFCLAWTDSLGSDTEQLVANILQWFAFGFSIL
                     ILMFYAYQTWRATCGWEEVYVCCVELTKVIIEFFHEFDDPSMLYLANGHRVQWLRYAEW
                     LLTCPVILIHLSNLTGLKDDYSKRTMRLLVSDVGTIVWGATSAMSTGYVKVIFFVLGCI
                     YGANTFFHAAKVYIESYHVVPKGRPRTVVRIMAWLFFLSWGMFPVLFVVGPEGFDAISV
                     YGSTIGHTIIDLMSKNCWGLLGHYLRVLIHQHIIIYGDIRKKTKINVAGEEMEVETMVD
                     QEDEETV"
     CDS             1745..2194
                     /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             join(2213..2215,2216..2926)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2213..2215,2216..2926)
                     /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(2255..2276)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2516..2535)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2863..2884
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2963..3551
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3016..3036)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3434..3445)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3584..3601)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3590..3797
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3806..3935
                     /label=AAV2 ITR
     rep_origin      4024..4479
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4111..4130)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4321..4342
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4546..4565)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4665..4687
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4725..4743)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4811..4915
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4916..4984,4985..5776)
                     /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(5134..5153)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5947..6535
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6436..6455
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6597..6607,1..119)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcgtcgg gcgacctttg gtcgcccggc
       61 ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctg
      121 cggccgcacg cgttctagaa atattaaggt acgggaggta cttggagcgg ccgcaataaa
      181 atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat agtactaaca
      241 tacgctctcc atcaaaacaa aacgaaacaa aacaaactag caaaataggc tgtccccagt
      301 gcaagtgcag gtgccagaac atttctctat tctagactcg agctgcagag ggccctgcgt
      361 atgagtgcaa gtgggtttta ggaccaggat gaggcggggt gggggtgcct acctgacgac
      421 cgaccccgac ccactggaca agcacccaac ccccattccc caaattgcgc atcccctatc
      481 agagaggggg aggggaaaca ggatgcggcg aggcgcgtgc gcactgccag cttcagcacc
      541 gcggacagtg ccttcgcccc cgcctggcgg cgcgcgccac cgccgcctca gcactgaagg
      601 cgcgctgacg tcactcgccg gtcccccgca aactcccctt cccggccacc ttggtcgcgt
      661 ccgcgccgcc gccggcccag ccggaccgca ccacgcgagg cgcgagatag gggggcacgg
      721 gcgcgaccat ctgcgctgcg gcgccggcga ctcagcgctg cctcagtctg cggtgggcag
      781 cggaggagtc gtgtcgtgcc tgagagcgca gtcgaattca agctgctagc aaggatccgc
      841 caccatgctg gggaacggca gcgccattgt gcctatcgac cagtgctttt gcctggcttg
      901 gaccgacagc ctgggaagcg atacagagca gctggtggcc aacatcctcc agtggttcgc
      961 cttcggcttc agcatcctga tcctgatgtt ctacgcctac cagacttgga gagccacttg
     1021 cggttgggag gaggtctacg tctgttgcgt cgagctgacc aaggtcatca tcgagttctt
     1081 ccacgagttc gacgacccca gcatgctgta cctggctaac ggacaccgag tccagtggct
     1141 gagatacgca gagtggctgc tgacttgtcc cgtcatcctg atccacctga gcaacctgac
     1201 cggcctgaag gacgactaca gcaagcggac catgaggctg ctggtgtcag acgtgggaac
     1261 catcgtgtgg ggagctacaa gcgccatgag cacaggctac gtcaaggtca tcttcttcgt
     1321 gctgggttgc atctacggcg ccaacacctt cttccacgcc gccaaggtgt atatcgagag
     1381 ctaccacgtg gtgccaaagg gcagacctag aaccgtcgtg cggatcatgg cttggctgtt
     1441 cttcctgtct tggggcatgt tccccgtgct gttcgtcgtg ggaccagaag gattcgacgc
     1501 catcagcgtg tacggctcta ccattggcca caccatcatc gacctcatga gcaagaattg
     1561 ttggggcctg ctgggacact atctgagagt gctgatccac cagcacatca tcatctacgg
     1621 cgacatccgg aagaagacca agatcaacgt ggccggcgag gagatggaag tggagaccat
     1681 ggtggaccag gaggacgagg agacagtggg aggttcaggt ggaaccggtg gaagtggagg
     1741 taccatgccc gccgaactgc tgctgctgct gattgtcgct tttgctaacc cttcttgcca
     1801 ggtgctgtct tctctgagga tggccgctat tctggacgat cagaccgtgt gcggaagagg
     1861 agagaggctg gcactggcac tggctaggga gcagatcaat ggcatcattg aagtgcctgc
     1921 aaaggcccgg gtcgaggtgg acattttcga actgcagaga gatagccagt acgagaccac
     1981 agacaccatg tgccagatcc tgccaaaagg agtggtctcc gtcctgggac caagctcctc
     2041 tcctgcttct gcaagtaccg tgtctcacat ttgtggcgag aaggaaatcc cccatatcaa
     2101 ggtcgggcca gaggaaacac ccaggctgca gtacctgcgc ttcgcctcag tgagcctgta
     2161 tccatcaaac gaggatgtgt cactggcagt cagcggagct agcggaggta ctgtgagcaa
     2221 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     2281 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     2341 cctgaagttc atttgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     2401 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     2461 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     2521 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     2581 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     2641 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     2701 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     2761 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     2821 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     2881 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagttct gctacgagaa
     2941 tgaagtgtaa aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     3001 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3061 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3121 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3181 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3241 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3301 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     3361 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgccctt
     3421 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3481 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3541 cgcctccccg catcgatacc gtcgactaga gctcgctgat cagcctcgac tgtgccttct
     3601 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     3661 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3721 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3781 agcaggcatg ctggggagag atctgaggaa cccctagtga tggagttggc cactccctct
     3841 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     3901 gcccgggcgg cctcagtgag cgagcgagcg cgcagagagg gagcccctgc agcctggcgt
     3961 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     4021 tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4081 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4141 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4201 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     4261 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4321 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     4381 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     4441 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct
     4501 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
     4561 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     4621 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     4681 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     4741 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     4801 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4861 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4921 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4981 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5041 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5101 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5161 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5221 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5281 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5341 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5401 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5461 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5521 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5581 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5641 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5701 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5761 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5821 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5881 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5941 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6001 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6061 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6121 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6181 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6241 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6301 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6361 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6421 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6481 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6541 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtcctg
     6601 caggcag
//
