LOCUS       sequence-247000-        6926 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 the CamKIIa promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6926)
  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 6926)
  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-ChRger2-TS-EYFP
FEATURES             Location/Qualifiers
     source          1..6926
                     /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(2594..2596,2597..3310)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2636..2657)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2897..2916)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3244..3265
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3338..3926
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3391..3411)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3809..3820)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3960..4071
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4189..4329
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4189..4318
                     /label=AAV2 ITR
     rep_origin      4404..4859
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4491..4510)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4701..4722
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4876..4895)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4995..5017
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5055..5073)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5141..5245
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5246..5314,5315..6106)
                     /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(5464..5483)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6277..6865
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6766..6785
                     /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 agtggcggcc gccaagagca ggatcaccag cgagggcgag tacatccccc tggaccagat
     2581 cgacatcaac gtggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     2641 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     2701 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     2761 ctggcccacc ctcgtgacca ccttcggcta cggcctgcag tgcttcgccc gctaccccga
     2821 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     2881 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     2941 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     3001 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     3061 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     3121 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     3181 cgacaaccac tacctgagct accagtccgc cctgagcaaa gaccccaacg agaagcgcga
     3241 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     3301 gtacaagtaa taagaattcg atatcaagct tatcgataat caacctctgg attacaaaat
     3361 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     3421 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     3481 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     3541 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     3601 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     3661 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     3721 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tatgttgcca cctggattct
     3781 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     3841 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     3901 gatctccctt tgggccgcct ccccgcatcg ataccgagcg ctgctcgaga gatcttcgac
     3961 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     4021 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     4081 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     4141 ggaagacaat agcaggcatg agatctcacg tgcggaccga gcggccgcag gaacccctag
     4201 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     4261 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     4321 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     4381 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4441 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     4501 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4561 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     4621 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4681 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     4741 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     4801 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     4861 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     4921 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     4981 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     5041 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     5101 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5161 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5221 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5281 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5341 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5401 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     5461 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     5521 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5581 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5641 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5701 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5761 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     5821 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5881 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5941 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     6001 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     6061 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     6121 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6181 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6241 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6301 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6361 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6421 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6481 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6541 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6601 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6661 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6721 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6781 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6841 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6901 cttttgctgg ccttttgctc acatgt
//
