LOCUS       sequence-247091-        6417 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High photocurrent, low-light sensitive channelrhodopsin (ChRger3) 
            for optogenetic activation with systemic delivery or for low-light 
            activation. Driven by human Synapsin I promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6417)
  AUTHORS   Bedbrook CN, Yang KK, Robinson JE, Mackey ED, Gradinaru V, Arnold FH
  TITLE     Machine learning-guided channelrhodopsin engineering enables 
            minimally invasive optogenetics.
  JOURNAL   Nat Methods. 2019 Oct 14. pii: 10.1038/s41592-019-0583-8. doi: 
            10.1038/s41592-019-0583-8.
  PUBMED    31611694
REFERENCE   2  (bases 1 to 6417)
  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"; journalName: "Nat 
            Methods. 2019 Oct 14. pii: 10.1038/s41592-019-0583-8. doi: 
            10.1038/s41592-019-0583-8."
COMMENT     Sequence Label: pAAV-hSyn-ChRger3-TS-EYFP
FEATURES             Location/Qualifiers
     source          1..6417
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             717..755
                     /codon_start=1
                     /product="peptide derived from the CnaB2 adhesin domain of 
                     Streptococcus pyogenes FbaB"
                     /label=SpyTag
                     /note="forms a covalent isopeptide bond with the SpyCatcher
                     protein (Zakeri et al., 2012)"
                     /translation="AHIVMVDAYKPTK"
     CDS             join(1800..1802,1803..2516)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1842..1863)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2103..2122)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2450..2471
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2544..3132
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2597..2617)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3015..3026)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3164..3640
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3363..3382)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3680..3820
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3680..3809
                     /label=AAV2 ITR
     rep_origin      3895..4350
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3982..4001)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4192..4213
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4367..4386)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4486..4508
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4546..4564)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4632..4736
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4737..4805,4806..5597)
                     /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(4955..4974)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5768..6356
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6257..6276
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc cgccaccatg tcgcggaggc
      661 catggcttct tgccctagcg ctggcagtgg cgctggcggc cggcagcgca ggagccgccc
      721 acatcgtgat ggtggacgcc tacaagccca ccaagtcgac tggcagtgac gcgacggtgc
      781 cggtcgcgac tcaggatggc cccgactacg ttttccaccg tgcccacgag cgcatgctct
      841 tccaaacctc atacactctt gagaacaatg gttctgttat ttgcatcccg aacaacggcc
      901 agtgcttctg cttggcttgg cttaaatcca acggaacaaa tgccgagaag ttggctgcca
      961 acattctgca gtggattact tttgcgcttt cagcgctctg cctgatgttc tacggctacc
     1021 agacctggaa gtctacttgc ggctgggagg agatttacgt ggccacgatc gagatgatca
     1081 agttcatcat cgagtatttc catgagtttg acagtcctgc gatgctgttc ctgtacggag
     1141 gcaacattac cccatggctt cgttacgccg agtggctgct gacatgccct gtcattctta
     1201 tccatctgag caacatcacc ggtctgagtg aggcatataa taagcgtacc atggctctgc
     1261 tggtgtcaga cctgggcact atctgcatgg gcgtgacagc cgctctggcc actgggtggg
     1321 tgaagtggct gttttactgt atcggcctgg tgtacggcac ccagacattc tacaacgcag
     1381 gaatcatcta cgtggaggcg taccacaccg tgccgaaggg ccggtgtcgc caggtggtga
     1441 ctggcatggc ttggctcttc ttcgtatcat ggggtatgtt ccccatcctg ttcatcctcg
     1501 gccccgaggg cttcggcgtc ctgagcgtgg ctgggtccac tattggccac accatcgccg
     1561 acctgctgtc caagaatatt tggggactgc tcgggcactt cctgcgcatc aaaatccacg
     1621 agcatatcct catccacggc gacattcgca agaccaccaa attgaacatt ggtggcactg
     1681 agattgaggt cgagacgctg gtggaggacg aggccgaggc tggcgcggtc gcggccgcca
     1741 agagcaggat caccagcgag ggcgagtaca tccccctgga ccagatcgac atcaacgtgg
     1801 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1861 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     1921 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1981 tgaccacctt cggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc
     2041 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     2101 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     2161 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     2221 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     2281 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     2341 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     2401 tgagctacca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     2461 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaataag
     2521 aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga
     2581 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     2641 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     2701 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     2761 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     2821 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     2881 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     2941 catcgtcctt tccttggctg ctcgcctatg ttgccacctg gattctgcgc gggacgtcct
     3001 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     3061 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     3121 ccgcctcccc gcatcgatac cgagcgctgc tcgagagatc tacgggtggc atccctgtga
     3181 cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt
     3241 cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg
     3301 gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg
     3361 ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc
     3421 ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg
     3481 catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc
     3541 caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct
     3601 gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac
     3661 gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     3721 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     3781 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
     3841 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     3901 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3961 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4021 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     4081 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     4141 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4201 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga
     4261 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4321 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     4381 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     4441 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     4501 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     4561 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     4621 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     4681 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     4741 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     4801 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     4861 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     4921 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     4981 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     5041 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     5101 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     5161 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     5221 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     5281 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     5341 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     5401 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     5461 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     5521 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     5581 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     5641 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5701 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5761 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5821 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5881 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     5941 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6001 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6061 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6121 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6181 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6241 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6301 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6361 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
