LOCUS       sequence-204182-        7948 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High efficiency channelrhodopsin, neuron specific promoter, red 
            fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7948)
  AUTHORS   Berndt A, Schoenenberger P, Mattis J, Tye KM, Deisseroth K, Hegemann
            P, Oertner TG
  TITLE     High-efficiency channelrhodopsins for fast neuronal stimulation at 
            low light levels.
  JOURNAL   Proc Natl Acad Sci U S A. 2011 May 3;108(18):7595-600. doi: 
            10.1073/pnas.1017210108. Epub 2011 Apr 19.
  PUBMED    21504945
REFERENCE   2  (bases 1 to 7948)
  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: 
            "10.1073/pnas.1017210108"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2011-05-3- 3"; volume: "108"; issue: "18"; pages: "7595-600"
COMMENT     Sequence Label: pAAV hSyn1 ChR2 ET-TC 2A tDimer
FEATURES             Location/Qualifiers
     source          1..7948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..474
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      494..503
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             500..1426
                     /codon_start=1
                     /product="N-terminal fragment of channelrhodopsin-2 from 
                     the alga Chlamydomonas reinhardtii (Nagel et al., 2003)"
                     /label=ChR2
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYATWLLTCPVILIHLSNLTGLSNDYSRRTMGLLVSDIGCIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     CDS             1436..1489
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     regulatory      2441..2450
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2445..2454
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2447..3124
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MVASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGTLIYKVKFRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFL"
     CDS             3167..3841
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGTLIYKVKFRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQA
                     LKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHHL
                     FL"
     misc_feature    3869..4457
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3922..3942)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4340..4351)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4460..4476)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4461..4477)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    4537..5013
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4736..4755)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5053..5193
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5053..5182
                     /label=AAV2 ITR
     rep_origin      5268..5723
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5355..5374)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5565..5586
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5740..5759)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5859..5881
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5919..5937)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6005..6109
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6110..6178,6179..6970)
                     /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(6328..6347)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7141..7729
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7630..7649
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   7791..7931
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7802..7931
                     /label=AAV2 ITR
ORIGIN
        1 ctagactgca gagggccctg cgtatgagtg caagtgggtt ttaggaccag gatgaggcgg
       61 ggtgggggtg cctacctgac gaccgacccc gacccactgg acaagcaccc aacccccatt
      121 ccccaaattg cgcatcccct atcagagagg gggaggggaa acaggatgcg gcgaggcgcg
      181 tgcgcactgc cagcttcagc accgcggaca gtgccttcgc ccccgcctgg cggcgcgcgc
      241 caccgccgcc tcagcactga aggcgcgctg acgtcactcg ccggtccccc gcaaactccc
      301 cttcccggcc accttggtcg cgtccgcgcc gccgccggcc cagccggacc gcaccacgcg
      361 aggcgcgaga taggggggca cgggcgcgac catctgcgct gcggcgccgg cgactcagcg
      421 ctgcctcagt ctgcggtggg cagcggagga gtcgtgtcgt gcctgagagc gcagtcgaga
      481 attcaagctg ctagccacca tggattatgg aggcgccctg agtgccgttg ggcgcgagct
      541 gctatttgta acgaacccag tagtcgtcaa tggctctgta cttgtgcctg aggaccagtg
      601 ttactgcgcg ggctggattg agtcgcgtgg cacaaacggt gcccaaacgg cgtcgaacgt
      661 gctgcaatgg cttgctgctg gcttctccat cctactgctt atgttttacg cctaccaaac
      721 atggaagtca acctgcggct gggaggagat ctatgtgtgc gctatcgaga tggtcaaggt
      781 gattctcgag ttcttcttcg agtttaagaa cccgtccatg ctgtatctag ccacaggcca
      841 ccgcgtccag tggttgcgtt acgccacgtg gcttctcacc tgcccggtca ttctcattca
      901 cctgtcaaac ctgacgggct tgtccaacga ctacagcagg cgcaccatgg gtctgcttgt
      961 gtctgatatt ggctgtattg tgtggggcgc cacttccgcc atggccaccg gatacgtcaa
     1021 ggtcatcttc ttctgcctgg gtctgtgtta tggtgctaac acgttctttc acgctgccaa
     1081 ggcctacatc gagggttacc acaccgtgcc gaagggccgg tgtcgccagg tggtgactgg
     1141 catggcttgg ctcttcttcg tatcatgggg tatgttcccc atcctgttca tcctcggccc
     1201 cgagggcttc ggcgtcctga gcgtgtacgg ctccaccgtc ggccacacca tcattgacct
     1261 gatgtcgaag aactgctggg gtctgctcgg ccactacctg cgcgtgctga tccacgagca
     1321 tatcctcatc cacggcgaca ttcgcaagac caccaaattg aacattggtg gcactgagat
     1381 tgaggtcgag acgctggtgg aggacgaggc cgaggctggc gcggtacccg cggccgaggg
     1441 cagaggaagt cttctaacat gcggtgacgt ggaggagaat cccggccctg caccgggatc
     1501 cggcatggac gtggtgaatc agctggtggc tgggggtcag ttccgggtgg tcaaggagcc
     1561 ccttggcttc gtgaaggtgc tgcagtgggt ctttgccatc ttcgcctttg ctacgtgtgg
     1621 cagctacacc ggggagcttc ggctgagcgt ggagtgtgcc aacaagacgg agagtgccct
     1681 caacatcgaa gttgaattcg agtacccctt caggctgcac caagtgtact ttgatgcacc
     1741 ctcctgcgtc aaagggggca ctaccaagat cttcctggtt ggggactact cctcgtcggc
     1801 tgaattcttt gtcaccgtgg ctgtgtttgc cttcctctac tccatggggg ccctggccac
     1861 ctacatcttc ctgcagaaca agtaccgaga gaacaacaaa gggcctatga tggactttct
     1921 ggctacagcc gtgttcgctt tcatgtggct agttagttca tcagcctggg ccaaaggcct
     1981 gtccgatgtg aagatggcca cggacccaga gaacattatc aaggagatgc ccatgtgccg
     2041 ccagacaggg aacacatgca aggaactgag ggaccctgtg acttcaggac tcaacacctc
     2101 agtggtgttt ggcttcctga acctggtgct ctgggttggc aacttatggt tcgtgttcaa
     2161 ggagacaggc tgggcagccc cattcatgcg cgcacctcca ggcgccccgg aaaagcaacc
     2221 agcacctggc gatgcctacg gcgatgcggg ctacgggcag ggccccggag gctatgggcc
     2281 ccaagactcc tacgggcctc agggtggtta tcaacccgat tacgggcagc cagccagcgg
     2341 tggcggtggc tacgggcctc agggcgacta tgggcagcaa ggctatggcc aacagggtgc
     2401 gcccacctcc ttctccaatc agatggcggc cggatccgcg gccgccatgg tggcctcctc
     2461 cgaggacgtc atcaaagagt tcatgcgctt caaggtgcgc atggagggct ccgtgaacgg
     2521 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     2581 caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccccca
     2641 gttccagtac ggctccaagg cgtacgtgaa gcaccccgcc gacatccccg actacaagaa
     2701 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
     2761 ggtgaccgtg acccaggact cctccctgca ggacggcacg ctgatctaca aggtgaagtt
     2821 ccgcggcacc aacttccccc ccgacggccc cgtaatgcag aagaagacca tgggctggga
     2881 ggcctccacc gagcgcctgt acccccgcga cggcgtgctg aagggcgaga tccaccaggc
     2941 cctgaagctg aaggacggcg gccactacct ggtggagttc aagaccatct acatggccaa
     3001 gaagcccgtg cagctgcccg gctactacta cgtggacacc aagctggaca tcacctccca
     3061 caacgaggac tacaccatcg tggaacagta cgagcgctcc gagggccgcc accacctgtt
     3121 cctggggcat ggcaccggca gcaccggcag cggcagctcc ggcaccgcct cctccgagga
     3181 cgtcatcaaa gagttcatgc gcttcaaggt gcgcatggag ggctccgtga acggccacga
     3241 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     3301 gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc cccagttcca
     3361 gtacggctcc aaggcgtacg tgaagcaccc cgccgacatc cccgactaca agaagctgtc
     3421 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
     3481 cgtgacccag gactcctccc tgcaggacgg cacgctgatc tacaaggtga agttccgcgg
     3541 caccaacttc ccccccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     3601 caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc gagatccacc aggccctgaa
     3661 gctgaaggac ggcggccact acctggtgga gttcaagacc atctacatgg ccaagaagcc
     3721 cgtgcagctg cccggctact actacgtgga caccaagctg gacatcacct cccacaacga
     3781 ggactacacc atcgtggaac agtacgagcg ctccgagggc cgccaccacc tgttcctgta
     3841 ggtcgacttc gagcaaaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag
     3901 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     3961 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     4021 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     4081 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     4141 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     4201 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     4261 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     4321 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     4381 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     4441 ttgggccgcc tccccgcatc gataccgtcg acctcgagac gcgatcgcgg cgcgccggcc
     4501 tctctggcct cgacctcgag tctagagatg atctacgggt ggcatccctg tgacccctcc
     4561 ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata
     4621 aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg
     4681 ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat
     4741 tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg
     4801 gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc
     4861 aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg
     4921 gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta
     4981 caggcgtgaa ccactgctcc cttccctgtc cttctgattt tgtaggtaac cacgtgcgga
     5041 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     5101 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     5161 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     5221 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     5281 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     5341 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     5401 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     5461 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     5521 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     5581 aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc
     5641 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     5701 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     5761 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     5821 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     5881 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     5941 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     6001 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     6061 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     6121 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     6181 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     6241 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     6301 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     6361 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     6421 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     6481 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     6541 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     6601 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     6661 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     6721 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     6781 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     6841 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     6901 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     6961 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     7021 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     7081 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     7141 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     7201 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     7261 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     7321 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     7381 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     7441 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     7501 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     7561 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     7621 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     7681 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     7741 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
     7801 gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt
     7861 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
     7921 taggggttcc tgcggccgca cgcgtcga
//
