LOCUS       sequence-252555-        6855 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  fusion protein of Gaussia luciferase variant slow burn, Volvox 
            channelrhodopsin-1, and enhanced yellow fluorescent protein 
            (luminopsin LMO3) for bioluminescent optogenetics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6855)
  AUTHORS   Petersen ED, Sharkey ED, Pal A, Shafau LO, Zenchak-Petersen J, Pena 
            AJ, Aggarwal A, Prakash M, Hochgeschwender U
  TITLE     Restoring Function After Severe Spinal Cord Injury Through 
            BioLuminescent-OptoGenetics.
  JOURNAL   Front Neurol. 2022 Jan 20;12:792643. doi: 10.3389/fneur.2021.792643
  PUBMED    35126293
REFERENCE   2  (bases 1 to 6855)
  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: "Front 
            Neurol."; date: "2022-01-20"; volume: "12:792643. doi"; pages: " 
            10.3389/fneur.2021.79264"
COMMENT     Sequence Label: pAAV-Hb9-LMO3 (sbGLuc-VChR1-EYFP)
FEATURES             Location/Qualifiers
     source          1..6855
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   5..134
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      626..635
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             632..1186
                     /codon_start=1
                     /product="enhanced variant of secreted Gaussia luciferase"
                     /label=Gaussia-Dura Luc
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGKKLPLEVLKELEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIVDIPEIPGFKDLEPLEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGGD"
     CDS             632..1186
                     /codon_start=1
                     /product="secreted Gaussia luciferase"
                     /label=hGLuc
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGKKLPLEVLKELEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIVDIPEIPGFKDLEPLEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGGD"
     regulatory      1226..1235
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2141..2143,2144..2857)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2183..2204)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2444..2463)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2791..2812
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2887..2928
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2938..2955
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2986..3574
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3039..3059)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3457..3468)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3606..4082
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3805..3824)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4122..4262
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4122..4251
                     /label=AAV2 ITR
     rep_origin      4337..4792
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4424..4443)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4634..4655
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4809..4828)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4928..4950
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4988..5006)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5074..5178
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5179..5247,5248..6039)
                     /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(5397..5416)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6210..6798
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6699..6718
                     /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 gcacgcgttg aataaattta agcaggctaa ttaatatata
      181 aactagccca atttgtcaag ttgatttgta ttttagttaa ttgtgaaagt aattaccaca
      241 tggtcaaatt aacagctttc tggaaatgac caagcctgag gttttatttc cttcctgggt
      301 gaagaaaatt catttttcca agctcttgat gtgatgaata aaagtcataa atctgggtga
      361 ttggtgcagg cagagtctaa atggcttcat atttcatttt aggtttaata gaaatattca
      421 tgctctgttt taatgaaatt aaattgaagg gggatggggc cagagtagtt agctgatgaa
      481 ttgacaaaaa ctaatcagct ttattgggaa acaggtttaa gggcacgggc gtgtcaataa
      541 cgctcagcct gaccccctct tccattagct aggcaggctg attagagaat tcggtaccgg
      601 atcctctaga gtcgagctca agcttgccac catgggagtc aaagttctgt ttgccctgat
      661 ctgcatcgct gtggccgagg ccaagcccac cgagaacaac gaagacttca acatcgtggc
      721 cgtggccagc aacttcgcga ccacggatct cgatgctgac cgcgggaagt tgcccggcaa
      781 gaagctgccg ctggaggtgc tcaaagagct ggaagccaat gcccggaaag ctggctgcac
      841 caggggctgt ctgatctgcc tgtcccacat caagtgcacg cccaagatga agaagttcat
      901 cccaggacgc tgccacacct acgaaggcga caaagagtcc gcacagggcg gcataggcga
      961 ggcgatcgtc gacattcctg agattcctgg gttcaaggac ttggagcccc tggagcagtt
     1021 catcgcacag gtcgatctgt gtgtggactg cacaactggc tgcctcaaag ggcttgccaa
     1081 cgtgcagtgt tctgacctgc tcaagaagtg gctgccgcaa cgctgtgcga cctttgccag
     1141 caagatccag ggccaggtgg acaagatcaa gggggccggt ggtgacggcg acccactagt
     1201 ccagtgtggt ggaattgccg gatccgccac catggattac cctgttgcta gaagcctcat
     1261 agttcgctac ccaaccgacc tcggaaacgg caccgtctgc atgccaagag gacagtgtta
     1321 ctgtgaaggt tggcttcgga gtcgcggcac ttccattgaa aagacaatag caattactct
     1381 tcagtgggta gtctttgctt tgtcagtggc ttgcctgggg tggtatgcgt atcaagcgtg
     1441 gcgagctacc tgcggatggg aggaggttta cgtagccttg atagaaatga tgaaaagcat
     1501 catcgaggcc ttccacgagt tcgacagccc tgcaacactg tggctgtctt cagggaacgg
     1561 cgtagtttgg atgcggtatg gcgaatggct cctcacctgc ccggtccttc tgatccatct
     1621 gagcaacctc acaggcctga aggacgatta tagcaaaagg actatgggcc tgttggtttc
     1681 tgatgtggga tgcatcgtgt ggggcgcaac cagcgccatg tgtacggggt ggacgaagat
     1741 cctgttcttc ctcatctcat tgagctatgg tatgtatacc tattttcatg ctgctaaagt
     1801 ttatatcgaa gcattccaca cagttccaaa agggatttgt cgagaactgg tccgagtgat
     1861 ggcctggaca ttctttgtgg cttggggaat gtttccagtc ctgtttctgc tgggcacgga
     1921 aggattcggt catatcagcc cttatggatc tgccattggg cactccatcc tcgacctgat
     1981 tgcaaagaac atgtggggtg tgctggggaa ttacctgcgc gtcaaaatcc acgagcacat
     2041 cctgttgtat ggcgacatca gaaagaagca gaaaattacg atcgccggcc aagagatgga
     2101 ggttgagaca ctggtggctg aagaggagga cgcggccgcc gtgagcaagg gcgaggagct
     2161 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     2221 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     2281 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct tcggctacgg
     2341 cctgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     2401 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     2461 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     2521 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     2581 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     2641 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     2701 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccgccct
     2761 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     2821 cgggatcact ctcggcatgg acgagctgta caagtaacat atgctctaga gggcccgcgg
     2881 ttcgaaggta agcctatccc taaccctctc ctcggtctcg attctacgcg taccggtcat
     2941 catcaccatc accattgagt ttaattcgat atcaagctta tcgataatca acctctggat
     3001 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     3061 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     3121 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     3181 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     3241 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     3301 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     3361 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc
     3421 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     3481 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     3541 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgagcgct gctcgagaga
     3601 tctacgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca
     3661 ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg
     3721 tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga
     3781 agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat
     3841 cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg
     3901 agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga
     3961 gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc
     4021 caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc
     4081 ttctgatttt gtaggtaacc acgtgcggac cgagcggccg caggaacccc tagtgatgga
     4141 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     4201 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     4261 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     4321 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4381 acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc
     4441 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4501 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     4561 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4621 acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc
     4681 tattcttttg atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg
     4741 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
     4801 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     4861 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     4921 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     4981 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     5041 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5101 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5161 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5221 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5281 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5341 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5401 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5461 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5521 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5581 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5641 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5701 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5761 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5821 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5881 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5941 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6001 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     6061 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6121 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6181 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6241 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6301 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6361 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6421 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6481 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6541 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6601 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6661 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6721 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6781 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6841 tggccttttg ctcac
//
