LOCUS       sequence-203116-        6817 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A somatic channelrhodopsin, which enables single cell, single 
            millisecond resolution optogenetics. CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6817)
  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 6817)
  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-CAG-soCoChR-GFP
FEATURES             Location/Qualifiers
     source          1..6817
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   35..164
                     /label=AAV2 ITR
     promoter        487..764
                     /label=chicken beta-actin promoter
     primer_bind     1044..1066
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1128..1147
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1186..2049
                     /codon_start=1
                     /product="high-photocurrent channelrhodopsin from 
                     Chloromonas oogama (Shemesh et al., 2017)"
                     /label=CoChR
                     /translation="MLGNGSAIVPIDQCFCLAWTDSLGSDTEQLVANILQWFAFGFSIL
                     ILMFYAYQTWRATCGWEEVYVCCVELTKVIIEFFHEFDDPSMLYLANGHRVQWLRYAEW
                     LLTCPVILIHLSNLTGLKDDYSKRTMRLLVSDVGTIVWGATSAMSTGYVKVIFFVLGCI
                     YGANTFFHAAKVYIESYHVVPKGRPRTVVRIMAWLFFLSWGMFPVLFVVGPEGFDAISV
                     YGSTIGHTIIDLMSKNCWGLLGHYLRVLIHQHIIIYGDIRKKTKINVAGEEMEVETMVD
                     QEDEETV"
     CDS             2086..2535
                     /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(2554..2556,2557..3270)
                     /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             join(2554..2556,2557..3267)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2596..2617)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2857..2876)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3204..3225
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3295..3883
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3348..3368)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3766..3777)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3926..4047
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3937..3956)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3991..4010
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4209..4338
                     /label=AAV2 ITR
     rep_origin      complement(4575..5163)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4655..4674)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5334..6125,6126..6194))
                     /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     5957..5976
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6195..6299)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6325..6780)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6462..6483)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6674..6693
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcgatcggtg cgggcctctt cgctattacg ccagctgcgc gctcgctcgc tcactgaggc
       61 cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg
      121 agcgcgcaga gagggagtgg ccaactccat cactaggggt tccttgtagt taatgattaa
      181 cccgccatgc tacttatcta cgtagccatg ctctaggaag atcgtaggtt aattaaccat
      241 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      301 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      361 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      421 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      481 ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac
      541 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg
      601 ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga
      661 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc
      721 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgacgc
      781 tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg
      841 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag
      901 cgcttggttt aatgacggct tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc
      961 cgggagggcc ctttgtgcgg ggggagcggc tcggggctgt ccgcgggggg acggctgcct
     1021 tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg gctctagagc
     1081 ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt
     1141 tattgtgctg tctcatcatt ttggcaaaga attggatccg ccaccatgct ggggaacggc
     1201 agcgccattg tgcctatcga ccagtgcttt tgcctggctt ggaccgacag cctgggaagc
     1261 gatacagagc agctggtggc caacatcctc cagtggttcg ccttcggctt cagcatcctg
     1321 atcctgatgt tctacgccta ccagacttgg agagccactt gcggttggga ggaggtctac
     1381 gtctgttgcg tcgagctgac caaggtcatc atcgagttct tccacgagtt cgacgacccc
     1441 agcatgctgt acctggctaa cggacaccga gtccagtggc tgagatacgc agagtggctg
     1501 ctgacttgtc ccgtcatcct gatccacctg agcaacctga ccggcctgaa ggacgactac
     1561 agcaagcgga ccatgaggct gctggtgtca gacgtgggaa ccatcgtgtg gggagctaca
     1621 agcgccatga gcacaggcta cgtcaaggtc atcttcttcg tgctgggttg catctacggc
     1681 gccaacacct tcttccacgc cgccaaggtg tatatcgaga gctaccacgt ggtgccaaag
     1741 ggcagaccta gaaccgtcgt gcggatcatg gcttggctgt tcttcctgtc ttggggcatg
     1801 ttccccgtgc tgttcgtcgt gggaccagaa ggattcgacg ccatcagcgt gtacggctct
     1861 accattggcc acaccatcat cgacctcatg agcaagaatt gttggggcct gctgggacac
     1921 tatctgagag tgctgatcca ccagcacatc atcatctacg gcgacatccg gaagaagacc
     1981 aagatcaacg tggccggcga ggagatggaa gtggagacca tggtggacca ggaggacgag
     2041 gagacagtgg gaggttcagg tggaaccggt ggaagtggag gtaccatgcc cgccgaactg
     2101 ctgctgctgc tgattgtcgc ttttgctaac ccttcttgcc aggtgctgtc ttctctgagg
     2161 atggccgcta ttctggacga tcagaccgtg tgcggaagag gagagaggct ggcactggca
     2221 ctggctaggg agcagatcaa tggcatcatt gaagtgcctg caaaggcccg ggtcgaggtg
     2281 gacattttcg aactgcagag agatagccag tacgagacca cagacaccat gtgccagatc
     2341 ctgccaaaag gagtggtctc cgtcctggga ccaagctcct ctcctgcttc tgcaagtacc
     2401 gtgtctcaca tttgtggcga gaaggaaatc ccccatatca aggtcgggcc agaggaaaca
     2461 cccaggctgc agtacctgcg cttcgcctca gtgagcctgt atccatcaaa cgaggatgtg
     2521 tcactggcag tcagcggagc tagcggaggt actgtgagca agggcgagga gctgttcacc
     2581 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     2641 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catttgcacc
     2701 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     2761 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     2821 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     2881 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     2941 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     3001 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     3061 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     3121 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     3181 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     3241 actctcggca tggacgagct gtacaagtaa gaattcgata tcaagcttat cgataatcaa
     3301 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     3361 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     3421 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     3481 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     3541 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     3601 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     3661 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     3721 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     3781 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     3841 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgaccc
     3901 gggcggccgc ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact
     3961 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     4021 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     4081 gttcaggggg agatgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa
     4141 atcgataagg atcttcctag agcatggcta cgtagataag tagcatggcg ggttaatcat
     4201 taactacaag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     4261 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     4321 gagcgagcga gcgcgcagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     4381 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     4441 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     4501 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4561 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4621 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4681 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4741 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4801 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4861 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4921 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4981 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     5041 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5101 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5161 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5221 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5281 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5341 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5401 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5461 gcaatgatac cgcgcgaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5521 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5581 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5641 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5701 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     5761 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     5821 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     5881 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     5941 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6001 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6061 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6121 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6181 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6241 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6301 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
     6361 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     6421 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     6481 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     6541 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     6601 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     6661 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     6721 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     6781 cccattcgcc attcaggctg cgcaactgtt gggaagg
//
