LOCUS       sequence-252558-        6926 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  fusion protein of Gaussia luciferase variant M23, improved slowChloC
            anion conducting ChR, and enhanced yellow fluorescent protein for 
            bioluminescent optogenetics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6926)
  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 6926)
  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-iLMO4 (GLucM23-iChloC-EYFP)
FEATURES             Location/Qualifiers
     source          1..6926
                     /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             1273..2199
                     /codon_start=1
                     /product="N-terminal fragment of channelrhodopsin-2 from 
                     the alga Chlamydomonas reinhardtii (Nagel et al., 2003)"
                     /label=ChR2
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEQIYVCAIRMVKVILEFFFSFKN
                     PSMLYLATGHRVQWLRYAEWLLTCPVILIHLSNLTGLSNDYSRRTMGLLVSNIGCIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     CDS             join(2212..2214,2215..2928)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2254..2275)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2515..2534)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2862..2883
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2958..2999
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3009..3026
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    3057..3645
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3110..3130)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3528..3539)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3677..4153
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3876..3895)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4193..4333
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4193..4322
                     /label=AAV2 ITR
     rep_origin      4408..4863
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4495..4514)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4705..4726
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4880..4899)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4999..5021
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5059..5077)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5145..5249
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5250..5318,5319..6110)
                     /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(5468..5487)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6281..6869
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6770..6789
                     /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 tggaggcgcc ctgagtgccg ttgggcgcga gctgctattt
     1321 gtaacgaacc cagtagtcgt caatggctct gtacttgtgc ctgaggacca gtgttactgc
     1381 gcgggctgga ttgagtcgcg tggcacaaac ggtgcccaaa cggcgtcgaa cgtgctgcaa
     1441 tggcttgctg ctggcttctc catcctactg cttatgtttt acgcctacca aacatggaag
     1501 tcaacctgcg gctgggagca gatctatgtg tgcgctatca ggatggtcaa ggtgattctc
     1561 gagttcttct tctcctttaa gaacccgtcc atgctgtatc tagccacagg ccaccgcgtc
     1621 cagtggttgc gttacgccga gtggcttctc acctgcccgg tcattctcat tcacctgtca
     1681 aacctgacgg gcttgtccaa cgactacagc aggcgcacca tgggtctgct tgtgtctaat
     1741 attggctgta ttgtgtgggg cgccacttcc gccatggcca ccggatacgt caaggtcatc
     1801 ttcttctgcc tgggtctgtg ttatggtgct aacacgttct ttcacgctgc caaggcctac
     1861 atcgagggtt accacaccgt gccgaagggc cggtgtcgcc aggtggtgac tggcatggct
     1921 tggctcttct tcgtatcatg gggtatgttc cccatcctgt tcatcctcgg ccccgagggc
     1981 ttcggcgtcc tgagcgtgta cggctccacc gtcggccaca ccatcattga cctgatgtcg
     2041 aagaactgct ggggtctgct cggccactac ctgcgcgtgc tgatccacga gcatatcctc
     2101 atccacggcg acattcgcaa gaccaccaaa ttgaacattg gtggcactga gattgaggtc
     2161 gagacgctgg tggaggacga ggccgaggct ggcgccgtac ccgcggccgc cgtgagcaag
     2221 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     2281 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     2341 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     2401 ttcggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
     2461 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     2521 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     2581 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     2641 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     2701 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     2761 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac
     2821 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     2881 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaca tatgctctag
     2941 agggcccgcg gttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc
     3001 gtaccggtca tcatcaccat caccattgag tttaattcga tatcaagctt atcgataatc
     3061 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     3121 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     3181 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     3241 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     3301 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     3361 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     3421 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     3481 atgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     3541 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     3601 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgagcgc
     3661 tgctcgagag atctacgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct
     3721 ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt
     3781 gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg
     3841 gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc
     3901 agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc
     3961 tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt
     4021 tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca
     4081 ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc
     4141 cttccctgtc cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc
     4201 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     4261 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     4321 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     4381 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     4441 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt
     4501 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     4561 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4621 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     4681 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact
     4741 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa
     4801 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     4861 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     4921 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     4981 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     5041 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     5101 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     5161 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     5221 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     5281 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     5341 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     5401 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     5461 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     5521 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     5581 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     5641 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     5701 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     5761 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     5821 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     5881 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5941 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     6001 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     6061 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     6121 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     6181 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     6241 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6301 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6361 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     6421 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6481 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6541 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6601 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6661 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6721 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6781 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6841 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6901 tggccttttg ctggcctttt gctcac
//
