LOCUS       sequence-247096-        6941 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 the CamKIIa promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6941)
  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 6941)
  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-CaMKIIa-ChRger3-TS-EYFP
FEATURES             Location/Qualifiers
     source          1..6941
                     /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        153..1441
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     CDS             1526..1564
                     /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(2609..2611,2612..3325)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2651..2672)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2912..2931)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3259..3280
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3353..3941
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3406..3426)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3824..3835)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3975..4086
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4204..4344
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4204..4333
                     /label=AAV2 ITR
     rep_origin      4419..4874
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4506..4525)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4716..4737
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4891..4910)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5010..5032
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5070..5088)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5156..5260
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5261..5329,5330..6121)
                     /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(5479..5498)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6292..6880
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6781..6800
                     /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 gcgtttaatt aacattatgg ccttaggtca cttcatctcc
      181 atggggttct tcttctgatt ttctagaaaa tgagatgggg gtgcagagag cttcctcagt
      241 gacctgccca gggtcacatc agaaatgtca gagctagaac ttgaactcag attactaatc
      301 ttaaattcca tgccttgggg gcatgcaagt acgatataca gaaggagtga actcattagg
      361 gcagatgacc aatgagttta ggaaagaaga gtccagggca gggtacatct acaccacccg
      421 cccagccctg ggtgagtcca gccacgttca cctcattata gttgcctctc tccagtccta
      481 ccttgacggg aagcacaagc agaaactggg acaggagccc caggagacca aatcttcatg
      541 gtccctctgg gaggatgggt ggggagagct gtggcagagg cctcaggagg ggccctgctg
      601 ctcagtggtg acagataggg gtgagaaagc agacagagtc attccgtcag cattctgggt
      661 ctgtttggta cttcttctca cgctaaggtg gcggtgtgat atgcacaatg gctaaaaagc
      721 agggagagct ggaaagaaac aaggacagag acagaggcca agtcaaccag accaattccc
      781 agaggaagca aagaaaccat tacagagact acaaggggga agggaaggag agatgaatta
      841 gcttcccctg taaaccttag aacccagctg ttgccagggc aacggggcaa tacctgtctc
      901 ttcagaggag atgaagttgc cagggtaact acatcctgtc tttctcaagg accatcccag
      961 aatgtggcac ccactagccg ttaccatagc aactgcctct ttgccccact taatcccatc
     1021 ccgtctgtta aaagggccct atagttggag gtgggggagg taggaagagc gatgatcact
     1081 tgtggactaa gtttgttcgc atccccttct ccaaccccct cagtacatca ccctggggga
     1141 acagggtcca cttgctcctg ggcccacaca gtcctgcagt attgtgtata taaggccagg
     1201 gcaaagagga gcaggtttta aagtgaaagg caggcaggtg ttggggaggc agttaccggg
     1261 gcaacgggaa cagggcgttt cggaggtggt tgccatgggg acctggatgc tgacgaaggc
     1321 tcgcgaggct gtgagcagcc acagtgccct gctcagaagc cccaagctcg tcagtcaagc
     1381 cggttctccg tttgcactca ggagcacggg caggcgagtg gcccctagtt ctgggggcag
     1441 cgggggatcc gccaccatgt cgcggaggcc atggcttctt gccctagcgc tggcagtggc
     1501 gctggcggcc ggcagcgcag gagccgccca catcgtgatg gtggacgcct acaagcccac
     1561 caagtcgact ggcagtgacg cgacggtgcc ggtcgcgact caggatggcc ccgactacgt
     1621 tttccaccgt gcccacgagc gcatgctctt ccaaacctca tacactcttg agaacaatgg
     1681 ttctgttatt tgcatcccga acaacggcca gtgcttctgc ttggcttggc ttaaatccaa
     1741 cggaacaaat gccgagaagt tggctgccaa cattctgcag tggattactt ttgcgctttc
     1801 agcgctctgc ctgatgttct acggctacca gacctggaag tctacttgcg gctgggagga
     1861 gatttacgtg gccacgatcg agatgatcaa gttcatcatc gagtatttcc atgagtttga
     1921 cagtcctgcg atgctgttcc tgtacggagg caacattacc ccatggcttc gttacgccga
     1981 gtggctgctg acatgccctg tcattcttat ccatctgagc aacatcaccg gtctgagtga
     2041 ggcatataat aagcgtacca tggctctgct ggtgtcagac ctgggcacta tctgcatggg
     2101 cgtgacagcc gctctggcca ctgggtgggt gaagtggctg ttttactgta tcggcctggt
     2161 gtacggcacc cagacattct acaacgcagg aatcatctac gtggaggcgt accacaccgt
     2221 gccgaagggc cggtgtcgcc aggtggtgac tggcatggct tggctcttct tcgtatcatg
     2281 gggtatgttc cccatcctgt tcatcctcgg ccccgagggc ttcggcgtcc tgagcgtggc
     2341 tgggtccact attggccaca ccatcgccga cctgctgtcc aagaatattt ggggactgct
     2401 cgggcacttc ctgcgcatca aaatccacga gcatatcctc atccacggcg acattcgcaa
     2461 gaccaccaaa ttgaacattg gtggcactga gattgaggtc gagacgctgg tggaggacga
     2521 ggccgaggct ggcgcggtcg cggccgccaa gagcaggatc accagcgagg gcgagtacat
     2581 ccccctggac cagatcgaca tcaacgtggt gagcaagggc gaggagctgt tcaccggggt
     2641 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     2701 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     2761 caagctgccc gtgccctggc ccaccctcgt gaccaccttc ggctacggcc tgcagtgctt
     2821 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     2881 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     2941 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     3001 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     3061 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
     3121 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     3181 ccccgtgctg ctgcccgaca accactacct gagctaccag tccgccctga gcaaagaccc
     3241 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     3301 cggcatggac gagctgtaca agtaataaga attcgatatc aagcttatcg ataatcaacc
     3361 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     3421 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     3481 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     3541 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     3601 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     3661 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     3721 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctatgt
     3781 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     3841 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     3901 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gagcgctgct
     3961 cgagagatct tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     4021 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     4081 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     4141 caagggggag gattgggaag acaatagcag gcatgagatc tcacgtgcgg accgagcggc
     4201 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     4261 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     4321 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     4381 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     4441 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     4501 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4561 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4621 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4681 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4741 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
     4801 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     4861 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     4921 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     4981 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     5041 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     5101 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     5161 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5221 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5281 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5341 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5401 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5461 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5521 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5581 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5641 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5701 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5761 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     5821 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     5881 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5941 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     6001 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6061 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6121 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     6181 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6241 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6301 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6361 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6421 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6481 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6541 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6601 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6661 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6721 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6781 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6841 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6901 tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
