LOCUS       sequence-216294-        7010 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 7010)
  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 7010)
  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..7010
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   5..145
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   16..145
                     /label=AAV2 ITR
     promoter        190..637
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             684..1238
                     /codon_start=1
                     /product="secreted Gaussia luciferase"
                     /label=hGLuc
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGKKLPLEVLKEMEANARKAGCTRGCLICLSHIKCTPKMKKWLPGRCHTYEGDKESA
                     QGGIGEAIVDIPEIPGFKDLEPMEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGGD"
     regulatory      1278..1287
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2400..2402,2403..3116)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2400..2402,2403..3113)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2442..2463)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2703..2722)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3050..3071
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3141..3729
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3194..3214)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3612..3623)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3761..4237
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3960..3979)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4277..4417
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4277..4406
                     /label=AAV2 ITR
     rep_origin      4492..4947
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4579..4598)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4789..4810
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4964..4983)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5083..5105
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5143..5161)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5229..5333
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5334..5402,5403..6194)
                     /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(5552..5571)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6365..6953
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6854..6873
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg
       61 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg
      121 gccaactcca tcactagggg ttcctgcggc cgcacgcgtg tgtctagact gcagagggcc
      181 ctgcgtatga gtgcaagtgg gttttaggac caggatgagg cggggtgggg gtgcctacct
      241 gacgaccgac cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc
      301 cctatcagag agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc
      361 agcaccgcgg acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac
      421 tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg
      481 tcgcgtccgc gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg
      541 gcacgggcgc gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt
      601 gggcagcgga ggagtcgtgt cgtgcctgag agcgcagtcg agaaggtacc ggatcctcta
      661 gagtcgagct caagcttggc accatgggag tcaaagttct gtttgccctg atctgcatcg
      721 ctgtggccga ggccaagccc accgagaaca acgaagactt caacatcgtg gccgtggcca
      781 gcaacttcgc gaccacggat ctcgatgctg accgcgggaa gttgcccggc aagaagctgc
      841 cgctggaggt gctcaaagag atggaagcca atgcccggaa agctggctgc accaggggct
      901 gtctgatctg cctgtcccac atcaagtgca cgcccaagat gaagaagtgg ctgccaggac
      961 gctgccacac ctacgaaggc gacaaagagt ccgcacaggg cggcataggc gaggcgatcg
     1021 tcgacattcc tgagattcct gggttcaagg acttggagcc catggagcag ttcatcgcac
     1081 aggtcgatct gtgtgtggac tgcacaactg gctgcctcaa agggcttgcc aacgtgcagt
     1141 gttctgacct gctcaagaag tggctgccgc aacgctgtgc gacctttgcc agcaagatcc
     1201 agggccaggt ggacaagatc aagggggccg gtggtgacgg cgacccacta gtccagtgtg
     1261 gtggaattgc cggatctgcc accatgggga cctggatgct gacgaaggct cgcgaggctg
     1321 tgagcagcca cagtgccctg ctcagaagcc ccaagctcgt caatcaagct ggttctccat
     1381 ttgcactcag gagcacgggc aggcgagtgg cccctagttc tgggggcagc gggggatccg
     1441 ccaccatgat cgtggaccag ttcgaggagg tgctgatgaa gaccagccag ctgttcccac
     1501 tgccaaccgc tacccagagc gcccagccaa cccacgtggc ccccgtgcca accgtgctgc
     1561 ccgacacccc catctacgag accgtgggcg acagcggcag caagaccctg tgggtggtgt
     1621 tcgtgctgat gctgatcgcc agcgccgcct tcaccgccct gagctggaag atccccgtga
     1681 acaggaggct gtaccacgtg atcaccacca tcatcaccct gaccgccgcc ctgagctact
     1741 tcgctatggc taccggccac ggagtggccc tgaacaagat cgtgatcagg acccagcacg
     1801 accacgtgcc cgacacctac gagaccgtgt accgacaggt gtactacgcc aggtacatcg
     1861 actgggctat caccacccca ctgctgctgc tggacctggg actgctggct ggaatgagcg
     1921 gagcccacat cttcatggcc atcgtggctg acctgatcat ggtgctgacc ggcctgttcg
     1981 ctgctttcgg cagcgaggga accccacaga agtggggatg gtacaccatc gcctgcatcg
     2041 cctacatctt cgtggtgtgg cacctggtgc tgaacggcgg cgccaacgcc agggtgaagg
     2101 gcgagaagct gaggagcttc ttcgtggcca tcggagctta caccctgatc ctgtggaccg
     2161 cttacccaat cgtgtgggga ctggctgacg gagctaggaa gatcggagtg gacggagaga
     2221 tcatcgctta cgctgtgctg gacgtgctgg ctaagggagt gttcggagct tggctgctgg
     2281 tgacccacgc caacctgagg gagagcgacg tggagctgaa cggcttctgg gccaacggcc
     2341 tgaacaggga gggcgccatc aggatcggcg aggacgacgg cgcccgaccg gtagtagcag
     2401 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     2461 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     2521 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     2581 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     2641 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     2701 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     2761 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     2821 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     2881 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     2941 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     3001 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     3061 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaagaat
     3121 tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg
     3181 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     3241 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     3301 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     3361 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     3421 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     3481 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     3541 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     3601 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     3661 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     3721 cctccccgca tcgataccga gcgctgctcg agagatctac gggtggcatc cctgtgaccc
     3781 ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct
     3841 aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg
     3901 gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt
     3961 ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc
     4021 ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat
     4081 gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag
     4141 gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg
     4201 attacaggcg tgaaccactg ctcccttccc tgtccttctg attttgtagg taaccacgtg
     4261 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     4321 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     4381 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     4441 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     4501 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4561 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4621 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4681 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     4741 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4801 tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta taagggattt
     4861 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4921 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     4981 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     5041 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     5101 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     5161 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     5221 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     5281 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     5341 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     5401 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     5461 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5521 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5581 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5641 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5701 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5761 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5821 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5881 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     5941 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     6001 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     6061 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     6121 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     6181 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     6241 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6301 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6361 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6421 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6481 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     6541 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6601 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6661 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6721 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6781 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6841 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6901 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6961 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
//
