LOCUS       sequence-246998-        6402 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High photocurrent, low-light sensitive channelrhodopsin (ChRger2) 
            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 6402)
  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 6402)
  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-ChRger2-TS-EYFP
FEATURES             Location/Qualifiers
     source          1..6402
                     /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             723..761
                     /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(1785..1787,1788..2501)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1827..1848)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2088..2107)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2435..2456
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2529..3117
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2582..2602)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3000..3011)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3149..3625
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3348..3367)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3665..3805
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3665..3794
                     /label=AAV2 ITR
     rep_origin      3880..4335
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3967..3986)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4177..4198
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4352..4371)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4471..4493
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4531..4549)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4617..4721
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4722..4790,4791..5582)
                     /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(4940..4959)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5753..6341
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6242..6261
                     /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 agcagactgg
      661 tcgccgcttc ttggctgctg gctctcctcc tctgcggaat taccagcaca acaacagcct
      721 ctgcccacat cgtgatggtg gacgcctaca agcccaccaa gagcgcccca gcagcttctt
      781 ctacagacgg aacagccgcc gcagcagtgt ctcactacgc catgaacggc ttcgacgagc
      841 tggctaaagg agccgtggtg ccagaagacc actttgtctg cggaccagcc gacaagtgct
      901 attgctccgc ttggctgcac agcagaggag ccctgtggga gcaggaaaca gccaggggct
      961 tccagtggtt tgccgtgttc ctgtctgccc tgtttctggc tttctacggc tggcacgcct
     1021 ataaggccag cgtgggctgg gaggaggtgt acgtgtgctc cgtggagctg atcaaagtga
     1081 ttctggagat ctatttcgag tttaccagtc ctgcgatgct gttcctgtac ggaggcaaca
     1141 ttaccccatg gcttcgttac gccgagtggc tgctgacatg ccctgtcatt cttatccatc
     1201 tgagcaacat caccggtctg agtgaggcat ataataagcg taccatggct ctgctggtgt
     1261 cagacctggg cactatctgc atgggcgtga cagccgctct ggccactggg tgggtgaagt
     1321 ggctgtttta ctgtatcggc ctggtgtacg gcacccagac attctacaac gcaggaatca
     1381 tctacgtgga ggcgtaccac accgtgccga agggccggtg tcgccaggtg gtgactggca
     1441 tggcttggct cttcttcgta tcatggggta tgttccccat cctgttcatc ctcggccccg
     1501 agggcttcgg cgtcctgagc gtggctgggt ccactattgg ccacaccatc gccgacctgc
     1561 tgtccaagaa tatttgggga ctgctcgggc acttcctgcg catcaaaatc cacgagcata
     1621 tcattatgta cggcgacatt cgcagaccag tgagctccca gtttctgggc cgcaaggtgg
     1681 acgtcctggc cttcgtgaca gaggaggata aagtggcggc cgccaagagc aggatcacca
     1741 gcgagggcga gtacatcccc ctggaccaga tcgacatcaa cgtggtgagc aagggcgagg
     1801 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     1861 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     1921 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accttcggct
     1981 acggcctgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     2041 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     2101 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     2161 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     2221 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     2281 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     2341 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtccg
     2401 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     2461 ccgccgggat cactctcggc atggacgagc tgtacaagta ataagaattc gatatcaagc
     2521 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     2581 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     2641 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     2701 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     2761 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     2821 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     2881 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     2941 ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     3001 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     3061 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc
     3121 gataccgagc gctgctcgag agatctacgg gtggcatccc tgtgacccct ccccagtgcc
     3181 tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag
     3241 ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt
     3301 atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc
     3361 aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc
     3421 gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag
     3481 ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa
     3541 ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg
     3601 aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg gaccgagcgg
     3661 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     3721 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     3781 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     3841 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     3901 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag
     3961 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4021 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4081 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4141 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4201 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     4261 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4321 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     4381 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     4441 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     4501 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     4561 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     4621 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4681 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4741 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4801 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4861 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     4921 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4981 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5041 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5101 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5161 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5221 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5281 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5341 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5401 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5461 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5521 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5581 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     5641 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5701 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5761 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5821 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5881 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5941 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6001 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6061 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6121 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6181 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6241 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6301 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6361 ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
