LOCUS       sequence-209345-        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   39..168
                     /label=AAV2 ITR
     promoter        241..1418
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          472..1409
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1366..1386
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1431..2294
                     /codon_start=1
                     /product="high-photocurrent channelrhodopsin from 
                     Chloromonas oogama (Shemesh et al., 2017)"
                     /label=CoChR
                     /translation="MLGNGSAIVPIDQCFCLAWTDSLGSDTEQLVANILQWFAFGFSIL
                     ILMFYAYQTWRATCGWEEVYVCCVELTKVIIEFFHEFDDPSMLYLANGHRVQWLRYAEW
                     LLTCPVILIHLSNLTGLKDDYSKRTMRLLVSDVGTIVWGATSAMSTGYVKVIFFVLGCI
                     YGANTFFHAAKVYIESYHVVPKGRPRTVVRIMAWLFFLSWGMFPVLFVVGPEGFDAISV
                     YGSTIGHTIIDLMSKNCWGLLGHYLRVLIHQHIIIYGDIRKKTKINVAGEEMEVETMVD
                     QEDEETV"
     CDS             2331..2780
                     /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    4251..4839
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4304..4324)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4722..4733)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4882..5003
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4893..4912)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4947..4966
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   5165..5294
                     /label=AAV2 ITR
     rep_origin      complement(5531..6119)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5611..5630)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6290..7081,7082..7150))
                     /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     6913..6932
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7151..7255)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7281..7736)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7418..7439)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7630..7649
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct gcgcgctcgc tcgctcactg
       61 aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg
      121 agcgagcgcg cagagaggga gtggccaact ccatcactag gggttccttg tagttaatga
      181 ttaacccgcc atgctactta tctacgtagc catgctctag gaagatcgta ggttaattaa
      241 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
      301 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      361 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      421 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      481 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      541 acttccactg gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg
      601 agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc
      661 ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt
      721 tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc
      781 aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg
      841 cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag
      901 cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg
      961 gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag
     1021 ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga
     1081 cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt
     1141 cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt
     1201 agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg
     1261 agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat
     1321 tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag
     1381 tggttcaaag tttttttctt ccatttcagg tgtcgtgagg atccgccacc atgctgggga
     1441 acggcagcgc cattgtgcct atcgaccagt gcttttgcct ggcttggacc gacagcctgg
     1501 gaagcgatac agagcagctg gtggccaaca tcctccagtg gttcgccttc ggcttcagca
     1561 tcctgatcct gatgttctac gcctaccaga cttggagagc cacttgcggt tgggaggagg
     1621 tctacgtctg ttgcgtcgag ctgaccaagg tcatcatcga gttcttccac gagttcgacg
     1681 accccagcat gctgtacctg gctaacggac accgagtcca gtggctgaga tacgcagagt
     1741 ggctgctgac ttgtcccgtc atcctgatcc acctgagcaa cctgaccggc ctgaaggacg
     1801 actacagcaa gcggaccatg aggctgctgg tgtcagacgt gggaaccatc gtgtggggag
     1861 ctacaagcgc catgagcaca ggctacgtca aggtcatctt cttcgtgctg ggttgcatct
     1921 acggcgccaa caccttcttc cacgccgcca aggtgtatat cgagagctac cacgtggtgc
     1981 caaagggcag acctagaacc gtcgtgcgga tcatggcttg gctgttcttc ctgtcttggg
     2041 gcatgttccc cgtgctgttc gtcgtgggac cagaaggatt cgacgccatc agcgtgtacg
     2101 gctctaccat tggccacacc atcatcgacc tcatgagcaa gaattgttgg ggcctgctgg
     2161 gacactatct gagagtgctg atccaccagc acatcatcat ctacggcgac atccggaaga
     2221 agaccaagat caacgtggcc ggcgaggaga tggaagtgga gaccatggtg gaccaggagg
     2281 acgaggagac agtgggaggt tcaggtggaa ccggtggaag tggaggtacc atgcccgccg
     2341 aactgctgct gctgctgatt gtcgcttttg ctaacccttc ttgccaggtg ctgtcttctc
     2401 tgaggatggc cgctattctg gacgatcaga ccgtgtgcgg aagaggagag aggctggcac
     2461 tggcactggc tagggagcag atcaatggca tcattgaagt gcctgcaaag gcccgggtcg
     2521 aggtggacat tttcgaactg cagagagata gccagtacga gaccacagac accatgtgcc
     2581 agatcctgcc aaaaggagtg gtctccgtcc tgggaccaag ctcctctcct gcttctgcaa
     2641 gtaccgtgtc tcacatttgt ggcgagaagg aaatccccca tatcaaggtc gggccagagg
     2701 aaacacccag gctgcagtac ctgcgcttcg cctcagtgag cctgtatcca tcaaacgagg
     2761 atgtgtcact ggcagtcagc ggagctagcg gaggtactgt gagtaagggc gaggaagtga
     2821 tcaaagagtt catgcggttt aaggtgagaa tggaaggaag catgaacggc cacgagttcg
     2881 aaattgaggg agaaggagag ggacggccct acgagggcac ccagacagcc aagctgaaag
     2941 tgacaaaggg cgggcctctg ccattcgctt gggacatcct gagcccacag tttatgggcg
     3001 gctccaaggc ctatgtgaaa catccagctg acattcccga ttataagaaa ctgagcttcc
     3061 ccgaggggtt taagtgggaa agagtgatga acttcgagga cggaggcctg gtgactgtga
     3121 cccaggacag ctccctgcag gatgggaccc tgatctacaa ggtgaaaatg agagggacaa
     3181 attttccccc tgatggacct gtgatgcaga agaaaactat gggatgggag gcctccaccg
     3241 aaaggctgta tccacgcgac ggggtgctga aaggagaaat ccaccaggct ctgaagctga
     3301 aagatggggg acattacctg gtggagttca agacaatcta catggccaag aaacctgtgc
     3361 agctgccagg ctactattac gtggacacaa aactggatat cacttcacac aacgaggact
     3421 acactattgt ggagcagtat gaacggagcg aggggagaca ccatctgttc ctgggccatg
     3481 ggactggaag taccggctca gggtctagtg gaaccgcctc aagcgaggat aacaatatgg
     3541 ctgtgatcaa agagttcatg aggtttaagg tgcgcatgga gggcagcatg aatgggcacg
     3601 aatttgagat tgaaggagag ggcgaaggga ggccttacga gggcacacag actgccaagc
     3661 tgaaagtgac caagggagga ccactgcctt tcgcttggga tatcctgtct cctcagttta
     3721 tgggcggaag taaggcctat gtcaagcatc ccgctgacat tcctgattac aagaaactgt
     3781 ctttcccaga gggctttaag tgggagagag tgatgaattt tgaagatgga ggcctggtga
     3841 ccgtgacaca ggactcctct ctgcaggatg gcactctgat ctacaaagtc aaaatgcgcg
     3901 gcaccaattt tccacccgat gggcccgtga tgcagaagaa aacaatgggg tgggaggcca
     3961 gcactgaacg gctgtatcct agagacggag tgctgaaggg cgaaatccac caggccctga
     4021 agctgaaaga cggcggccac tacctggtgg agttcaaaac catctacatg gccaagaaac
     4081 cagtgcagct gcccggctat tactatgtgg acaccaagct ggatatcaca tcccacaatg
     4141 aagactacac cattgtggaa cagtatgaga ggtctgaagg acgccaccat ctgtttctgt
     4201 acggcatgga tgagctgtat aagtaagaat tcgatatcaa gcttatcgat aatcaacctc
     4261 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     4321 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     4381 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     4441 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     4501 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     4561 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     4621 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg
     4681 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     4741 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     4801 ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacccgggc
     4861 ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     4921 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     4981 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     5041 agggggagat gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg
     5101 ataaggatct tcctagagca tggctacgta gataagtagc atggcgggtt aatcattaac
     5161 tacaaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     5221 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     5281 gagcgagcgc gcagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5341 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5401 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5461 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5521 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5581 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5641 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5701 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5761 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5821 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5881 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5941 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     6001 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6061 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6121 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6181 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6241 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6301 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6361 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6421 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6481 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6541 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6601 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6661 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6721 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6781 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6841 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6901 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6961 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7021 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7081 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7141 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7201 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7261 ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     7321 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     7381 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     7441 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     7501 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     7561 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     7621 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     7681 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca
     7741 ttcgccattc aggctgcgca actgttggg
//
