LOCUS       sequence-252513-        7086 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  fusion protein of Gaussia luciferase variant superluminescent, 
            Leptosphaeria maculans opsin, and enhanced yellow fluorescent 
            protein for bioluminescent optogenetics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7086)
  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 7086)
  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-iLMO (slGLuc-Mac-EYFP)
FEATURES             Location/Qualifiers
     source          1..7086
                     /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      652..661
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             658..1212
                     /codon_start=1
                     /product="secreted Gaussia luciferase"
                     /label=hGLuc
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGKKLPLEVLKEMEANARKAGCTRGCLICLSHIKCTPKMKKWLPGRCHTYEGDKESA
                     QGGIGEAIVDIPEIPGFKDLEPMEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGGD"
     regulatory      1252..1261
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2452..2454,2455..3165)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2494..2515)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2755..2774)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3102..3123
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3217..3805
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3270..3290)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3688..3699)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3837..4313
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4036..4055)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4353..4493
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4353..4482
                     /label=AAV2 ITR
     rep_origin      4568..5023
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4655..4674)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4865..4886
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5040..5059)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5159..5181
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5219..5237)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5305..5409
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5410..5478,5479..6270)
                     /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(5628..5647)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6441..7029
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6930..6949
                     /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 gatccaagct tgccaccatg
      661 ggagtcaaag ttctgtttgc cctgatctgc atcgctgtgg ccgaggccaa gcccaccgag
      721 aacaacgaag acttcaacat cgtggccgtg gccagcaact tcgcgaccac ggatctcgat
      781 gctgaccgcg ggaagttgcc cggcaagaag ctgccgctgg aggtgctcaa agagatggaa
      841 gccaatgccc ggaaagctgg ctgcaccagg ggctgtctga tctgcctgtc ccacatcaag
      901 tgcacgccca agatgaagaa gtggctgcca ggacgctgcc acacctacga aggcgacaaa
      961 gagtccgcac agggcggcat aggcgaggcg atcgtcgaca ttcctgagat tcctgggttc
     1021 aaggacttgg agcccatgga gcagttcatc gcacaggtcg atctgtgtgt ggactgcaca
     1081 actggctgcc tcaaagggct tgccaacgtg cagtgttctg acctgctcaa gaagtggctg
     1141 ccgcaacgct gtgcgacctt tgccagcaag atccagggcc aggtggacaa gatcaagggg
     1201 gccggtggtg acggcgaccc actagtccag tgtggtggaa ttgccggatc tgccaccatg
     1261 gggacctgga tgctgacgaa ggctcgcgag gctgtgagca gccacagtgc cctgctcaga
     1321 agccccaagc tcgtcaatca agctggttct ccatttgcac tcaggagcac gggcaggcga
     1381 gtggccccta gttctggggg cagcggggga tccgccacca tgatcgtgga ccagttcgag
     1441 gaggtgctga tgaagaccag ccagctgttc ccactgccaa ccgctaccca gagcgcccag
     1501 ccaacccacg tggcccccgt gccaaccgtg ctgcccgaca cccccatcta cgagaccgtg
     1561 ggcgacagcg gcagcaagac cctgtgggtg gtgttcgtgc tgatgctgat cgccagcgcc
     1621 gccttcaccg ccctgagctg gaagatcccc gtgaacagga ggctgtacca cgtgatcacc
     1681 accatcatca ccctgaccgc cgccctgagc tacttcgcta tggctaccgg ccacggagtg
     1741 gccctgaaca agatcgtgat caggacccag cacgaccacg tgcccgacac ctacgagacc
     1801 gtgtaccgac aggtgtacta cgccaggtac atcgactggg ctatcaccac cccactgctg
     1861 ctgctggacc tgggactgct ggctggaatg agcggagccc acatcttcat ggccatcgtg
     1921 gctgacctga tcatggtgct gaccggcctg ttcgctgctt tcggcagcga gggaacccca
     1981 cagaagtggg gatggtacac catcgcctgc atcgcctaca tcttcgtggt gtggcacctg
     2041 gtgctgaacg gcggcgccaa cgccagggtg aagggcgaga agctgaggag cttcttcgtg
     2101 gccatcggag cttacaccct gatcctgtgg accgcttacc caatcgtgtg gggactggct
     2161 gacggagcta ggaagatcgg agtggacgga gagatcatcg cttacgctgt gctggacgtg
     2221 ctggctaagg gagtgttcgg agcttggctg ctggtgaccc acgccaacct gagggagagc
     2281 gacgtggagc tgaacggctt ctgggccaac ggcctgaaca gggagggcgc catcaggatc
     2341 ggcgaggacg acggcgcccg accggtagta gcagtgagca aggcggccgc caagagcagg
     2401 atcaccagcg agggcgagta catccccctg gaccagatcg acatcaacgt ggtgagcaag
     2461 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     2521 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     2581 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     2641 ttcggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
     2701 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     2761 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     2821 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     2881 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     2941 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     3001 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac
     3061 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     3121 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagttctg ctacgagaac
     3181 gaggtgtaat gagaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt
     3241 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     3301 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     3361 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     3421 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     3481 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     3541 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     3601 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg
     3661 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     3721 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     3781 atctcccttt gggccgcctc cccgcatcga taccgagcgc tgctcgagag atctacgggt
     3841 ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc
     3901 ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct
     3961 ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct
     4021 gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca
     4081 ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg
     4141 gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt
     4201 tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc
     4261 ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt
     4321 tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac
     4381 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     4441 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
     4501 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     4561 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4621 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
     4681 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4741 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     4801 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4861 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
     4921 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
     4981 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     5041 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     5101 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     5161 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     5221 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     5281 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     5341 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     5401 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     5461 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     5521 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     5581 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     5641 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5701 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     5761 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5821 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5881 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5941 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     6001 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     6061 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     6121 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     6181 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     6241 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     6301 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6361 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6421 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6481 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6541 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6601 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6661 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6721 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6781 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6841 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6901 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6961 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7021 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7081 gctcac
//
