LOCUS       sequence-252568-        6937 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  fusion protein of Gaussia luciferase variant slow burn, 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 6937)
  AUTHORS   Berglund K, Clissold K, Li HE, Wen L, Park SY, Gleixner J, Klein ME,
            Lu D, Barter JW, Rossi MA, Augustine GJ, Yin HH, Hochgeschwender U
  TITLE     Luminopsins integrate opto- and chemogenetics by using physical and 
            biological light sources for opsin activation.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 Jan 19;113(3):E358-67. doi: 
            10.1073/pnas.1510899113. Epub 2016 Jan 5.
  PUBMED    26733686
REFERENCE   2  (bases 1 to 6937)
  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"; doi: 
            "10.1073/pnas.1510899113"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2016-01-19- 19"; volume: "113"; issue: "3"; pages: "E358-67"
COMMENT     Sequence Label: pAAV-hSyn-LMO3 (sbGLuc-VChR1-EYFP)
FEATURES             Location/Qualifiers
     source          1..6937
                     /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      687..696
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             693..1247
                     /codon_start=1
                     /product="enhanced variant of secreted Gaussia luciferase"
                     /label=Gaussia-Dura Luc
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGKKLPLEVLKELEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIVDIPEIPGFKDLEPLEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGGD"
     CDS             693..1247
                     /codon_start=1
                     /product="secreted Gaussia luciferase"
                     /label=hGLuc
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGKKLPLEVLKELEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIVDIPEIPGFKDLEPLEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGGD"
     regulatory      1287..1296
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2262..2264,2265..2975)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2304..2325)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2565..2584)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2912..2933
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3020..3037
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    3068..3656
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3121..3141)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3539..3550)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3688..4164
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3887..3906)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     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 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 accggtggcg cgccaccggt aagcttgcca ccatgggagt caaagttctg tttgccctga
      721 tctgcatcgc tgtggccgag gccaagccca ccgagaacaa cgaagacttc aacatcgtgg
      781 ccgtggccag caacttcgcg accacggatc tcgatgctga ccgcgggaag ttgcccggca
      841 agaagctgcc gctggaggtg ctcaaagagc tggaagccaa tgcccggaaa gctggctgca
      901 ccaggggctg tctgatctgc ctgtcccaca tcaagtgcac gcccaagatg aagaagttca
      961 tcccaggacg ctgccacacc tacgaaggcg acaaagagtc cgcacagggc ggcataggcg
     1021 aggcgatcgt cgacattcct gagattcctg ggttcaagga cttggagccc ctggagcagt
     1081 tcatcgcaca ggtcgatctg tgtgtggact gcacaactgg ctgcctcaaa gggcttgcca
     1141 acgtgcagtg ttctgacctg ctcaagaagt ggctgccgca acgctgtgcg acctttgcca
     1201 gcaagatcca gggccaggtg gacaagatca agggggccgg tggtgacggc gacccactag
     1261 tccagtgtgg tggaattgcc ggatccgcca ccatggatta ccctgttgct agaagcctca
     1321 tagttcgcta cccaaccgac ctcggaaacg gcaccgtctg catgccaaga ggacagtgtt
     1381 actgtgaagg ttggcttcgg agtcgcggca cttccattga aaagacaata gcaattactc
     1441 ttcagtgggt agtctttgct ttgtcagtgg cttgcctggg gtggtatgcg tatcaagcgt
     1501 ggcgagctac ctgcggatgg gaggaggttt acgtagcctt gatagaaatg atgaaaagca
     1561 tcatcgaggc cttccacgag ttcgacagcc ctgcaacact gtggctgtct tcagggaacg
     1621 gcgtagtttg gatgcggtat ggcgaatggc tcctcacctg cccggtcctt ctgatccatc
     1681 tgagcaacct cacaggcctg aaggacgatt atagcaaaag gactatgggc ctgttggttt
     1741 ctgatgtggg atgcatcgtg tggggcgcaa ccagcgccat gtgtacgggg tggacgaaga
     1801 tcctgttctt cctcatctca ttgagctatg gtatgtatac ctattttcat gctgctaaag
     1861 tttatatcga agcattccac acagttccaa aagggatttg tcgagaactg gtccgagtga
     1921 tggcctggac attctttgtg gcttggggaa tgtttccagt cctgtttctg ctgggcacgg
     1981 aaggattcgg tcatatcagc ccttatggat ctgccattgg gcactccatc ctcgacctga
     2041 ttgcaaagaa catgtggggt gtgctgggga attacctgcg cgtcaaaatc cacgagcaca
     2101 tcctgttgta tggcgacatc agaaagaagc agaaaattac gatcgccggc caagagatgg
     2161 aggttgagac actggtggct gaagaggagg acgcggccgc caagagcagg atcaccagcg
     2221 agggcgagta catccccctg gaccagatcg acatcaacgt ggtgagcaag ggcgaggagc
     2281 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     2341 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     2401 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ttcggctacg
     2461 gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     2521 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     2581 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     2641 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     2701 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     2761 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     2821 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccgccc
     2881 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     2941 ccgggatcac tctcggcatg gacgagctgt acaagttctg ctacgagaac gaggtgtaat
     3001 gagctagcca attgccggtc atcatcacca tcaccattga gtttaattcg atatcaagct
     3061 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     3121 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     3181 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     3241 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     3301 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     3361 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     3421 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     3481 gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     3541 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     3601 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     3661 ataccgagcg ctgctcgaga gatctacggg tggcatccct gtgacccctc cccagtgcct
     3721 ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt
     3781 tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta
     3841 tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca
     3901 agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg
     3961 attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc
     4021 taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac
     4081 tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga
     4141 accactgctc ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg 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 tgctcac
//
