LOCUS       sequence-259728-        7495 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. Uses the CAG promoter and 
            double floxed..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7495)
  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 7495)
  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-CAG-DIO-(STChRger2-TS-EYFP)
FEATURES             Location/Qualifiers
     source          1..7495
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        210..513
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        515..792
                     /label=chicken beta-actin promoter
     primer_bind     1122..1144
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1206..1225
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(1292..1325)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(1376..1409)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(1421..2134,2135..2137))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1466..1487)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1815..1834
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2074..2095
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(3356..3394)
                     /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"
     protein_bind    3480..3513
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    3564..3597
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3622..4210
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3675..3695)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4093..4104)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4242..4718
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4441..4460)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4758..4898
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4758..4887
                     /label=AAV2 ITR
     rep_origin      4973..5428
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5060..5079)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5270..5291
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5445..5464)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5564..5586
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5624..5642)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5710..5814
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5815..5883,5884..6675)
                     /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(6033..6052)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6846..7434
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7335..7354
                     /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 gcgtaacact agttattaat agtaatcaat tacggggtca
      181 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      241 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      301 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      361 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      421 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      481 tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc
      541 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta
      601 ttttgtgcag cgatgggggc gggggggggg ggggggcgcg cgccgggggg gggggggggg
      661 ggsggggggs ggggsggggs gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg
      721 ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc
      781 gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc
      841 tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac
      901 ggcccttctc ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg
      961 tggctgcgtg aaagccttga ggggctccgg gagcgccggc aggaaggaaa tgggcgggga
     1021 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1081 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1141 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1201 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tgcggcccaa
     1261 cggtaccgga tcctctagag tcgactccgg aataacttcg tataggatac tttatacgaa
     1321 gttatgcaga atggtagctg gattgtagct gctattagca atatgaaacc tcttaataac
     1381 ttcgtatagc atacattata cgaagttatg gcgcgcccta ttacttgtac agctcgtcca
     1441 tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct
     1501 cgttggggtc tttgctcagg gcggactggt agctcaggta gtggttgtcg ggcagcagca
     1561 cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt
     1621 cctcgatgtt gtggcggatc ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca
     1681 tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt
     1741 cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
     1801 tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga
     1861 cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc
     1921 gggcgaagca ctgcaggccg tagccgaagg tggtcacgag ggtgggccag ggcacgggca
     1981 gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc
     2041 cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg
     2101 gcaccacccc ggtgaacagc tcctcgccct tgctcaccac gttgatgtcg atctggtcca
     2161 gggggatgta ctcgccctcg ctggtgatcc tgctcttggc ggccgccacg aatctggttg
     2221 cttcgggaaa gtcagttgca caactaatga agctatctat tgaactcatg gatcgcatgt
     2281 caatcacgcc ctcggttctg gctgggaggg gtgccactgg tgatggcatg gatgacatct
     2341 ccaactcttc tggtggttta ctctgtggag ccatctcctt agtattcaag ataggctgag
     2401 attgtttatc ctcctctgtc acgaaggcca ggacgtccac cttgcggccc agaaactggg
     2461 agctcactgg tctgcgaatg tcgccgtaca taatgatatg ctcgtggatt ttgatgcgca
     2521 ggaagtgccc gagcagtccc caaatattct tggacagcag gtcggcgatg gtgtggccaa
     2581 tagtggaccc agccacgctc aggacgccga agccctcggg gccgaggatg aacaggatgg
     2641 ggaacatacc ccatgatacg aagaagagcc aagccatgcc agtcaccacc tggcgacacc
     2701 ggcccttcgg cacggtgtgg tacgcctcca cgtagatgat tcctgcgttg tagaatgtct
     2761 gggtgccgta caccaggccg atacagtaaa acagccactt cacccaccca gtggccagag
     2821 cggctgtcac gcccatgcag atagtgccca ggtctgacac cagcagagcc atggtacgct
     2881 tattatatgc ctcactcaga ccggtgatgt tgctcagatg gataagaatg acagggcatg
     2941 tcagcagcca ctcggcgtaa cgaagccatg gggtaatgtt gcctccgtac aggaacagca
     3001 tcgcaggact ggtaaactcg aaatagatct ccagaatcac tttgatcagc tccacggagc
     3061 acacgtacac ctcctcccag cccacgctgg ccttataggc gtgccagccg tagaaagcca
     3121 gaaacagggc agacaggaac acggcaaacc actggaagcc cctggctgtt tcctgctccc
     3181 acagggctcc tctgctgtgc agccaagcgg agcaatagca cttgtcggct ggtccgcaga
     3241 caaagtggtc ttctggcacc acggctcctt tagccagctc gtcgaagccg ttcatggcgt
     3301 agtgagacac tgctgcggcg gctgttccgt ctgtagaaga agctgctggg gcgctcttgg
     3361 tgggcttgta ggcgtccacc atcacgatgt gggcagaggc tgttgttgtg ctggtaattc
     3421 cgcagaggag gagagccagc agccaagaag cggcgaccag tctgctcatg gtggctagca
     3481 taacttcgta taaagtatcc tatacgaagt tatttgcctt aacccagaaa ttatcactgt
     3541 tattctttag aatggtgcaa agaataactt cgtataatgt atgctatacg aagttatgaa
     3601 ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
     3661 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     3721 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     3781 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     3841 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     3901 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     3961 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     4021 tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc
     4081 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     4141 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     4201 gcctccccgc atcgataccg agcgctgctc gagagatcta cgggtggcat ccctgtgacc
     4261 cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc
     4321 taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt
     4381 ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg
     4441 tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct
     4501 cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca
     4561 tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca
     4621 ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg
     4681 gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttgtag gtaaccacgt
     4741 gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     4801 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     4861 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     4921 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     4981 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     5041 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     5101 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     5161 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     5221 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     5281 ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt
     5341 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     5401 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     5461 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     5521 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     5581 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     5641 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     5701 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5761 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     5821 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5881 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5941 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     6001 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     6061 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     6121 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     6181 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     6241 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     6301 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     6361 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     6421 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     6481 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     6541 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     6601 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     6661 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     6721 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6781 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6841 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6901 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6961 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     7021 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     7081 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     7141 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     7201 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     7261 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     7321 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     7381 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     7441 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
