LOCUS       sequence-203120-        7769 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A somatic channelrhodopsin, which enables single cell, single 
            millisecond resolution optogenetics. EF1alpha promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7769)
  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 7769)
  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-EF1alpha-soCoChR-tdTomato
FEATURES             Location/Qualifiers
     source          1..7769
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   35..164
                     /label=AAV2 ITR
     promoter        237..1414
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          468..1405
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1362..1382
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1427..2290
                     /codon_start=1
                     /product="high-photocurrent channelrhodopsin from 
                     Chloromonas oogama (Shemesh et al., 2017)"
                     /label=CoChR
                     /translation="MLGNGSAIVPIDQCFCLAWTDSLGSDTEQLVANILQWFAFGFSIL
                     ILMFYAYQTWRATCGWEEVYVCCVELTKVIIEFFHEFDDPSMLYLANGHRVQWLRYAEW
                     LLTCPVILIHLSNLTGLKDDYSKRTMRLLVSDVGTIVWGATSAMSTGYVKVIFFVLGCI
                     YGANTFFHAAKVYIESYHVVPKGRPRTVVRIMAWLFFLSWGMFPVLFVVGPEGFDAISV
                     YGSTIGHTIIDLMSKNCWGLLGHYLRVLIHQHIIIYGDIRKKTKINVAGEEMEVETMVD
                     QEDEETV"
     CDS             2327..2776
                     /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"
     misc_feature    4247..4835
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4300..4320)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4718..4729)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4878..4999
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4889..4908)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4943..4962
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   5161..5290
                     /label=AAV2 ITR
     rep_origin      complement(5527..6115)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5607..5626)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6286..7077,7078..7146))
                     /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     6909..6928
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7147..7251)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7277..7732)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7414..7435)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7626..7645
                     /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 aattaaggct
      241 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
      301 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
      361 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
      421 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt
      481 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt
      541 ccactggctg cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag
      601 ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct tgagttgagg cctggcctgg
      661 gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga
      721 taagtctcta gccatttaaa atttttgatg acctgctgcg acgctttttt tctggcaaga
      781 tagtcttgta aatgcgggcc aagatctgca cactggtatt tcggtttttg gggccgcggg
      841 cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg
      901 gccaccgaga atcggacggg ggtagtctca agctggccgg cctgctctgg tgcctggcct
      961 cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg caccagttgc
     1021 gtgagcggaa agatggccgc ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg
     1081 gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc
     1141 agccgtcgct tcatgtgact ccacggagta ccgggcgccg tccaggcacc tcgattagtt
     1201 ctcgagcttt tggagtacgt cgtctttagg ttggggggag gggttttatg cgatggagtt
     1261 tccccacact gagtgggtgg agactgaagt taggccagct tggcacttga tgtaattctc
     1321 cttggaattt gccctttttg agtttggatc ttggttcatt ctcaagcctc agacagtggt
     1381 tcaaagtttt tttcttccat ttcaggtgtc gtgaggatcc gccaccatgc tggggaacgg
     1441 cagcgccatt gtgcctatcg accagtgctt ttgcctggct tggaccgaca gcctgggaag
     1501 cgatacagag cagctggtgg ccaacatcct ccagtggttc gccttcggct tcagcatcct
     1561 gatcctgatg ttctacgcct accagacttg gagagccact tgcggttggg aggaggtcta
     1621 cgtctgttgc gtcgagctga ccaaggtcat catcgagttc ttccacgagt tcgacgaccc
     1681 cagcatgctg tacctggcta acggacaccg agtccagtgg ctgagatacg cagagtggct
     1741 gctgacttgt cccgtcatcc tgatccacct gagcaacctg accggcctga aggacgacta
     1801 cagcaagcgg accatgaggc tgctggtgtc agacgtggga accatcgtgt ggggagctac
     1861 aagcgccatg agcacaggct acgtcaaggt catcttcttc gtgctgggtt gcatctacgg
     1921 cgccaacacc ttcttccacg ccgccaaggt gtatatcgag agctaccacg tggtgccaaa
     1981 gggcagacct agaaccgtcg tgcggatcat ggcttggctg ttcttcctgt cttggggcat
     2041 gttccccgtg ctgttcgtcg tgggaccaga aggattcgac gccatcagcg tgtacggctc
     2101 taccattggc cacaccatca tcgacctcat gagcaagaat tgttggggcc tgctgggaca
     2161 ctatctgaga gtgctgatcc accagcacat catcatctac ggcgacatcc ggaagaagac
     2221 caagatcaac gtggccggcg aggagatgga agtggagacc atggtggacc aggaggacga
     2281 ggagacagtg ggaggttcag gtggaaccgg tggaagtgga ggtaccatgc ccgccgaact
     2341 gctgctgctg ctgattgtcg cttttgctaa cccttcttgc caggtgctgt cttctctgag
     2401 gatggccgct attctggacg atcagaccgt gtgcggaaga ggagagaggc tggcactggc
     2461 actggctagg gagcagatca atggcatcat tgaagtgcct gcaaaggccc gggtcgaggt
     2521 ggacattttc gaactgcaga gagatagcca gtacgagacc acagacacca tgtgccagat
     2581 cctgccaaaa ggagtggtct ccgtcctggg accaagctcc tctcctgctt ctgcaagtac
     2641 cgtgtctcac atttgtggcg agaaggaaat cccccatatc aaggtcgggc cagaggaaac
     2701 acccaggctg cagtacctgc gcttcgcctc agtgagcctg tatccatcaa acgaggatgt
     2761 gtcactggca gtcagcggag ctagcggagg tactgtgagt aagggcgagg aagtgatcaa
     2821 agagttcatg cggtttaagg tgagaatgga aggaagcatg aacggccacg agttcgaaat
     2881 tgagggagaa ggagagggac ggccctacga gggcacccag acagccaagc tgaaagtgac
     2941 aaagggcggg cctctgccat tcgcttggga catcctgagc ccacagttta tgggcggctc
     3001 caaggcctat gtgaaacatc cagctgacat tcccgattat aagaaactga gcttccccga
     3061 ggggtttaag tgggaaagag tgatgaactt cgaggacgga ggcctggtga ctgtgaccca
     3121 ggacagctcc ctgcaggatg ggaccctgat ctacaaggtg aaaatgagag ggacaaattt
     3181 tccccctgat ggacctgtga tgcagaagaa aactatggga tgggaggcct ccaccgaaag
     3241 gctgtatcca cgcgacgggg tgctgaaagg agaaatccac caggctctga agctgaaaga
     3301 tgggggacat tacctggtgg agttcaagac aatctacatg gccaagaaac ctgtgcagct
     3361 gccaggctac tattacgtgg acacaaaact ggatatcact tcacacaacg aggactacac
     3421 tattgtggag cagtatgaac ggagcgaggg gagacaccat ctgttcctgg gccatgggac
     3481 tggaagtacc ggctcagggt ctagtggaac cgcctcaagc gaggataaca atatggctgt
     3541 gatcaaagag ttcatgaggt ttaaggtgcg catggagggc agcatgaatg ggcacgaatt
     3601 tgagattgaa ggagagggcg aagggaggcc ttacgagggc acacagactg ccaagctgaa
     3661 agtgaccaag ggaggaccac tgcctttcgc ttgggatatc ctgtctcctc agtttatggg
     3721 cggaagtaag gcctatgtca agcatcccgc tgacattcct gattacaaga aactgtcttt
     3781 cccagagggc tttaagtggg agagagtgat gaattttgaa gatggaggcc tggtgaccgt
     3841 gacacaggac tcctctctgc aggatggcac tctgatctac aaagtcaaaa tgcgcggcac
     3901 caattttcca cccgatgggc ccgtgatgca gaagaaaaca atggggtggg aggccagcac
     3961 tgaacggctg tatcctagag acggagtgct gaagggcgaa atccaccagg ccctgaagct
     4021 gaaagacggc ggccactacc tggtggagtt caaaaccatc tacatggcca agaaaccagt
     4081 gcagctgccc ggctattact atgtggacac caagctggat atcacatccc acaatgaaga
     4141 ctacaccatt gtggaacagt atgagaggtc tgaaggacgc caccatctgt ttctgtacgg
     4201 catggatgag ctgtataagt aagaattcga tatcaagctt atcgataatc aacctctgga
     4261 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     4321 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     4381 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     4441 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     4501 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     4561 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     4621 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     4681 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     4741 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     4801 gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgtcgac ccgggcggcc
     4861 gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     4921 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     4981 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     5041 ggagatgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa
     5101 ggatcttcct agagcatggc tacgtagata agtagcatgg cgggttaatc attaactaca
     5161 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     5221 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     5281 gagcgcgcag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5341 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5401 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5461 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5521 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5581 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5641 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5701 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5761 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5821 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5881 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5941 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6001 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6061 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6121 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6181 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6241 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6301 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6361 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6421 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6481 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6541 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6601 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6661 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6721 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6781 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6841 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6901 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6961 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7021 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7081 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7141 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7201 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7261 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     7321 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     7381 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     7441 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     7501 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     7561 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     7621 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     7681 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg
     7741 ccattcaggc tgcgcaactg ttgggaagg
//
