LOCUS       sequence-252522-        6896 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  fusion protein of Gaussia luciferase variant M23, Volvox 
            channelrhodopsin-1, and enhanced yellow fluorescent protein for 
            bioluminescent optogenetics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6896)
  AUTHORS   Park SY, Song SH, Palmateer B, Pal A, Petersen ED, Shall GP, Welchko
            RM, Ibata K, Miyawaki A, Augustine GJ, Hochgeschwender U
  TITLE     Novel luciferase-opsin combinations for improved luminopsins.
  JOURNAL   J Neurosci Res. 2017 Sep 1. doi: 10.1002/jnr.24152.
  PUBMED    28862809
REFERENCE   2  (bases 1 to 6896)
  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: "J Neurosci 
            Res. 2017 Sep 1. doi: 10.1002/jnr.24152."
COMMENT     Sequence Label: pAAV-hSyn-LMO4 (GLucM23-VChR1-EYFP)
FEATURES             Location/Qualifiers
     source          1..6896
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   5..134
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        179..626
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      667..676
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             673..1227
                     /codon_start=1
                     /product="enhanced variant of secreted Gaussia luciferase"
                     /label=Gaussia-Dura Luc
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGEKLPLEVLKELEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIDDIPEIPGFKDLEPIEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGDD"
     CDS             673..1227
                     /codon_start=1
                     /product="secreted Gaussia luciferase"
                     /label=hGLuc
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGEKLPLEVLKELEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIDDIPEIPGFKDLEPIEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGDD"
     regulatory      1267..1276
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2182..2184,2185..2898)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2224..2245)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2485..2504)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2832..2853
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2928..2969
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2979..2996
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    3027..3615
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3080..3100)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3498..3509)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3647..4123
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3846..3865)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4163..4303
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4163..4292
                     /label=AAV2 ITR
     rep_origin      4378..4833
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4465..4484)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4675..4696
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4850..4869)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4969..4991
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5029..5047)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5115..5219
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5220..5288,5289..6080)
                     /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(5438..5457)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6251..6839
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6740..6759
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc gtcgggcgac
       61 ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat
      121 cactaggggt tcctgcggcc gcacgcgtgt gtctagactg cagagggccc tgcgtatgag
      181 tgcaagtggg ttttaggacc aggatgaggc ggggtggggg tgcctacctg acgaccgacc
      241 ccgacccact ggacaagcac ccaaccccca ttccccaaat tgcgcatccc ctatcagaga
      301 gggggagggg aaacaggatg cggcgaggcg cgtgcgcact gccagcttca gcaccgcgga
      361 cagtgccttc gcccccgcct ggcggcgcgc gccaccgccg cctcagcact gaaggcgcgc
      421 tgacgtcact cgccggtccc ccgcaaactc cccttcccgg ccaccttggt cgcgtccgcg
      481 ccgccgccgg cccagccgga ccgcaccacg cgaggcgcga gatagggggg cacgggcgcg
      541 accatctgcg ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg ggcagcggag
      601 gagtcgtgtc gtgcctgaga gcgcagtcga gaaggtaccg gatcctctag agtcgagctc
      661 aagcttgcca ccatgggagt caaagttctg tttgccctga tctgcatcgc tgtggccgag
      721 gccaagccca ccgagaacaa cgaagacttc aacatcgtgg ccgtggccag caacttcgcg
      781 accacggatc tcgatgctga ccgcgggaag ttgcccggcg agaagctgcc gctggaggtg
      841 ctcaaagagt tggaagccaa tgcccggaaa gctggctgca ccaggggctg tctgatctgc
      901 ctgtcccaca tcaagtgcac gcccaagatg aagaagttca tcccaggacg ctgccacacc
      961 tacgaaggcg acaaagagtc cgcacagggc ggcataggcg aggcgatcga cgacattcct
     1021 gagattcctg ggttcaagga cttggagccc atagagcagt tcatcgcaca ggtcgatctg
     1081 tgtgtggact gcacaactgg ctgcctcaaa gggcttgcca acgtgcagtg ttctgacctg
     1141 ctcaagaagt ggctgccgca acgctgcgcg acctttgcca gcaagatcca gggccaggtg
     1201 gacaagatca agggggccgg tgatgacggc gacccactag tccagtgtgg tggaattgcc
     1261 ggatccgcca ccatggatta ccctgttgct agaagcctca tagttcgcta cccaaccgac
     1321 ctcggaaacg gcaccgtctg catgccaaga ggacagtgtt actgtgaagg ttggcttcgg
     1381 agtcgcggca cttccattga aaagacaata gcaattactc ttcagtgggt agtctttgct
     1441 ttgtcagtgg cttgcctggg gtggtatgcg tatcaagcgt ggcgagctac ctgcggatgg
     1501 gaggaggttt acgtagcctt gatagaaatg atgaaaagca tcatcgaggc cttccacgag
     1561 ttcgacagcc ctgcaacact gtggctgtct tcagggaacg gcgtagtttg gatgcggtat
     1621 ggcgaatggc tcctcacctg cccggtcctt ctgatccatc tgagcaacct cacaggcctg
     1681 aaggacgatt atagcaaaag gactatgggc ctgttggttt ctgatgtggg atgcatcgtg
     1741 tggggcgcaa ccagcgccat gtgtacgggg tggacgaaga tcctgttctt cctcatctca
     1801 ttgagctatg gtatgtatac ctattttcat gctgctaaag tttatatcga agcattccac
     1861 acagttccaa aagggatttg tcgagaactg gtccgagtga tggcctggac attctttgtg
     1921 gcttggggaa tgtttccagt cctgtttctg ctgggcacgg aaggattcgg tcatatcagc
     1981 ccttatggat ctgccattgg gcactccatc ctcgacctga ttgcaaagaa catgtggggt
     2041 gtgctgggga attacctgcg cgtcaaaatc cacgagcaca tcctgttgta tggcgacatc
     2101 agaaagaagc agaaaattac gatcgccggc caagagatgg aggttgagac actggtggct
     2161 gaagaggagg acgcggccgc cgtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     2221 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     2281 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     2341 cccgtgccct ggcccaccct cgtgaccacc ttcggctacg gcctgcagtg cttcgcccgc
     2401 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     2461 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     2521 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     2581 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     2641 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     2701 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     2761 ctgctgcccg acaaccacta cctgagctac cagtccgccc tgagcaaaga ccccaacgag
     2821 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     2881 gacgagctgt acaagtaaca tatgctctag agggcccgcg gttcgaaggt aagcctatcc
     2941 ctaaccctct cctcggtctc gattctacgc gtaccggtca tcatcaccat caccattgag
     3001 tttaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
     3061 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     3121 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     3181 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     3241 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     3301 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     3361 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     3421 aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt
     3481 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     3541 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     3601 gggccgcctc cccgcatcga taccgagcgc tgctcgagag atctacgggt ggcatccctg
     3661 tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct
     3721 tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat
     3781 ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg
     3841 cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc
     3901 cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc
     3961 atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt
     4021 ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt
     4081 gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt tgtaggtaac
     4141 cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     4201 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     4261 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     4321 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     4381 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4441 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4501 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4561 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     4621 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4681 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag
     4741 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4801 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     4861 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     4921 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     4981 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     5041 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     5101 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5161 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     5221 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5281 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     5341 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5401 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5461 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5521 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5581 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5641 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5701 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5761 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5821 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5881 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5941 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     6001 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     6061 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     6121 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     6181 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     6241 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6301 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6361 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     6421 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6481 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6541 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6601 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6661 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6721 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6781 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6841 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcac
//
