LOCUS       sequence-258245-        7121 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Soma Targeted, High photocurrent, low-light sensitive 
            channelrhodopsin (ChRger2) 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 7121)
  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 7121)
  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-CaMKII-STChRger2-TS-EYFP
FEATURES             Location/Qualifiers
     source          1..7121
                     /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             1532..1570
                     /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(2789..2791,2792..3505)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGYVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(3092..3111)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3439..3460
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3533..4121
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3586..3606)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4004..4015)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4155..4266
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4384..4524
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4384..4513
                     /label=AAV2 ITR
     rep_origin      4599..5054
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4686..4705)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4896..4917
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5071..5090)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5190..5212
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5250..5268)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5336..5440
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5441..5509,5510..6301)
                     /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(5659..5678)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6472..7060
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6961..6980
                     /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 gccaccatga gcagactggt cgccgcttct tggctgctgg ctctcctcct
     1501 ctgcggaatt accagcacaa caacagcctc tgcccacatc gtgatggtgg acgcctacaa
     1561 gcccaccaag agcgccccag cagcttcttc tacagacgga acagccgccg cagcagtgtc
     1621 tcactacgcc atgaacggct tcgacgagct ggctaaagga gccgtggtgc cagaagacca
     1681 ctttgtctgc ggaccagccg acaagtgcta ttgctccgct tggctgcaca gcagaggagc
     1741 cctgtgggag caggaaacag ccaggggctt ccagtggttt gccgtgttcc tgtctgccct
     1801 gtttctggct ttctacggct ggcacgccta taaggccagc gtgggctggg aggaggtgta
     1861 cgtgtgctcc gtggagctga tcaaagtgat tctggagatc tatttcgagt ttaccagtcc
     1921 tgcgatgctg ttcctgtacg gaggcaacat taccccatgg cttcgttacg ccgagtggct
     1981 gctgacatgc cctgtcattc ttatccatct gagcaacatc accggtctga gtgaggcata
     2041 taataagcgt accatggctc tgctggtgtc agacctgggc actatctgca tgggcgtgac
     2101 agccgctctg gccactgggt gggtgaagtg gctgttttac tgtatcggcc tggtgtacgg
     2161 cacccagaca ttctacaacg caggaatcat ctacgtggag gcgtaccaca ccgtgccgaa
     2221 gggccggtgt cgccaggtgg tgactggcat ggcttggctc ttcttcgtat catggggtat
     2281 gttccccatc ctgttcatcc tcggccccga gggcttcggc gtcctgagcg tggctgggtc
     2341 cactattggc cacaccatcg ccgacctgct gtccaagaat atttggggac tgctcgggca
     2401 cttcctgcgc atcaaaatcc acgagcatat cattatgtac ggcgacattc gcagaccagt
     2461 gagctcccag tttctgggcc gcaaggtgga cgtcctggcc ttcgtgacag aggaggataa
     2521 acaatctcag cctatcttga atactaagga gatggctcca cagagtaaac caccagaaga
     2581 gttggagatg tcatccatgc catcaccagt ggcacccctc ccagccagaa ccgagggcgt
     2641 gattgacatg cgatccatga gttcaataga tagcttcatt agttgtgcaa ctgactttcc
     2701 cgaagcaacc agattcgtgg cggccgccaa gagcaggatc accagcgagg gcgagtacat
     2761 ccccctggac cagatcgaca tcaacgtggt gagcaagggc gaggagctgt tcaccggggt
     2821 ggtgcccatc ctggtcgagc tggacggcta cgtaaacggc cacaagttca gcgtgtccgg
     2881 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     2941 caagctgccc gtgccctggc ccaccctcgt gaccaccttc ggctacggcc tgcagtgctt
     3001 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     3061 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     3121 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     3181 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     3241 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
     3301 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     3361 ccccgtgctg ctgcccgaca accactacct gagctaccag tccgccctga gcaaagaccc
     3421 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     3481 cggcatggac gagctgtaca agtaataaga attcgatatc aagcttatcg ataatcaacc
     3541 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     3601 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     3661 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     3721 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     3781 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     3841 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     3901 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctatgt
     3961 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     4021 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     4081 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gagcgctgct
     4141 cgagagatct tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     4201 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     4261 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     4321 caagggggag gattgggaag acaatagcag gcatgagatc tcacgtgcgg accgagcggc
     4381 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     4441 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     4501 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     4561 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     4621 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     4681 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4741 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4801 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4861 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4921 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
     4981 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     5041 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     5101 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     5161 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     5221 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     5281 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     5341 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5401 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5461 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5521 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5581 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5641 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5701 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5761 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5821 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5881 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5941 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     6001 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     6061 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     6121 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     6181 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6241 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6301 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     6361 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6421 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6481 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6541 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6601 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6661 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6721 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6781 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6841 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6901 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6961 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     7021 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     7081 tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
