LOCUS       sequence-209402-        6777 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A somatic channelrhodopsin, which enables single cell, single 
            millisecond resolution optogenetics. Cre-dependent virus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6777)
  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 6777)
  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-FLEX-soCoChR-GFP
FEATURES             Location/Qualifiers
     source          1..6777
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   4..133
                     /label=AAV2 ITR
     promoter        236..683
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    699..732
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    795..828
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(850..1563,1564..1566))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             complement(join(853..1563,1564..1566))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(895..916)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1244..1263
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1503..1524
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(1585..2034)
                     /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(2071..2934)
                     /codon_start=1
                     /product="high-photocurrent channelrhodopsin from 
                     Chloromonas oogama (Shemesh et al., 2017)"
                     /label=CoChR
                     /translation="MLGNGSAIVPIDQCFCLAWTDSLGSDTEQLVANILQWFAFGFSIL
                     ILMFYAYQTWRATCGWEEVYVCCVELTKVIIEFFHEFDDPSMLYLANGHRVQWLRYAEW
                     LLTCPVILIHLSNLTGLKDDYSKRTMRLLVSDVGTIVWGATSAMSTGYVKVIFFVLGCI
                     YGANTFFHAAKVYIESYHVVPKGRPRTVVRIMAWLFFLSWGMFPVLFVVGPEGFDAISV
                     YGSTIGHTIIDLMSKNCWGLLGHYLRVLIHQHIIIYGDIRKKTKINVAGEEMEVETMVD
                     QEDEETV"
     protein_bind    complement(2962..2995)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(3066..3099)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3132..3720
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3185..3205)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3603..3614)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3761..3968
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4138..4267
                     /label=AAV2 ITR
     rep_origin      4339..4794
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4426..4445)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4636..4657
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4820..4924
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4925..4993,4994..5785)
                     /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(5143..5162)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5956..6544
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6445..6464
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
       61 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
      121 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
      181 tctaggaaga tcgtaggtta attaagtgtc tagactgcag agggccctgc gtatgagtgc
      241 aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg accgaccccg
      301 acccactgga caagcaccca acccccattc cccaaattgc gcatccccta tcagagaggg
      361 ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca ccgcggacag
      421 tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa ggcgcgctga
      481 cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc gtccgcgccg
      541 ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac gggcgcgacc
      601 atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc agcggaggag
      661 tcgtgtcgtg cctgagagcg cagtcgagaa gaattcccat aacttcgtat aaagtatcct
      721 atacgaagtt atatcaaaat aggaagacca atgcttcacc atcgacccga attgccaagc
      781 atcaccatcg acccataact tcgtataatg tatgctatac gaagttatac tagctagcgt
      841 ttaaactcat tacttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa
      901 ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcaggg cggactgggt
      961 gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg
     1021 gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg tggcggatct tgaagttcac
     1081 cttgatgccg ttcttctgct tgtcggccat gatatagacg ttgtggctgt tgtagttgta
     1141 ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat
     1201 gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc
     1261 gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa
     1321 gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt aggtcagggt
     1381 ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcaaatga acttcagggt
     1441 cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt
     1501 tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt
     1561 gctcacagta cctccgctag ctccgctgac tgccagtgac acatcctcgt ttgatggata
     1621 caggctcact gaggcgaagc gcaggtactg cagcctgggt gtttcctctg gcccgacctt
     1681 gatatggggg atttccttct cgccacaaat gtgagacacg gtacttgcag aagcaggaga
     1741 ggagcttggt cccaggacgg agaccactcc ttttggcagg atctggcaca tggtgtctgt
     1801 ggtctcgtac tggctatctc tctgcagttc gaaaatgtcc acctcgaccc gggcctttgc
     1861 aggcacttca atgatgccat tgatctgctc cctagccagt gccagtgcca gcctctctcc
     1921 tcttccgcac acggtctgat cgtccagaat agcggccatc ctcagagaag acagcacctg
     1981 gcaagaaggg ttagcaaaag cgacaatcag cagcagcagc agttcggcgg gcatggtacc
     2041 tccacttcca ccggttccac ctgaacctcc cactgtctcc tcgtcctcct ggtccaccat
     2101 ggtctccact tccatctcct cgccggccac gttgatcttg gtcttcttcc ggatgtcgcc
     2161 gtagatgatg atgtgctggt ggatcagcac tctcagatag tgtcccagca ggccccaaca
     2221 attcttgctc atgaggtcga tgatggtgtg gccaatggta gagccgtaca cgctgatggc
     2281 gtcgaatcct tctggtccca cgacgaacag cacggggaac atgccccaag acaggaagaa
     2341 cagccaagcc atgatccgca cgacggttct aggtctgccc tttggcacca cgtggtagct
     2401 ctcgatatac accttggcgg cgtggaagaa ggtgttggcg ccgtagatgc aacccagcac
     2461 gaagaagatg accttgacgt agcctgtgct catggcgctt gtagctcccc acacgatggt
     2521 tcccacgtct gacaccagca gcctcatggt ccgcttgctg tagtcgtcct tcaggccggt
     2581 caggttgctc aggtggatca ggatgacggg acaagtcagc agccactctg cgtatctcag
     2641 ccactggact cggtgtccgt tagccaggta cagcatgctg gggtcgtcga actcgtggaa
     2701 gaactcgatg atgaccttgg tcagctcgac gcaacagacg tagacctcct cccaaccgca
     2761 agtggctctc caagtctggt aggcgtagaa catcaggatc aggatgctga agccgaaggc
     2821 gaaccactgg aggatgttgg ccaccagctg ctctgtatcg cttcccaggc tgtcggtcca
     2881 agccaggcaa aagcactggt cgataggcac aatggcgctg ccgttcccca gcatggtggc
     2941 ggcgcggcct aggtacctag tataacttcg tataggatac tttatacgaa gttatcattg
     3001 ggattcttcc tattttgatc caagcatcac catcgaccct ctagtccaga tctcaccatc
     3061 gacccataac ttcgtatagc atacattata cgaagttatg tccctcgaag aggttcgcga
     3121 agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     3181 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     3241 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     3301 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     3361 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     3421 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     3481 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     3541 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     3601 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     3661 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     3721 atcgataccg tcgacccggg cggccgctca taaagctcga ctgtgccttc tagttgccag
     3781 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     3841 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     3901 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     3961 gctggggatg cggtccggac tgtacaattg cattcatttt atgtttcagg ttcaggggga
     4021 gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga
     4081 tcttcctaga gcatggctac gtagataagt agcatggcgg gttaatcatt aactacaagg
     4141 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     4201 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     4261 cgcgcagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     4321 gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4381 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     4441 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4501 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     4561 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4621 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     4681 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     4741 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     4801 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4861 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4921 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4981 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5041 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5101 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5161 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5221 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5281 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5341 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5401 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5461 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5521 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5581 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5641 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5701 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5761 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     5821 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5881 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5941 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6001 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6061 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6121 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6181 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6241 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6301 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6361 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6421 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6481 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6541 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6601 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     6661 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     6721 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatg
//
