LOCUS       sequence-252489-        7671 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  fusion protein of Gaussia luciferase, Volvox channelrhodopsin-1, and
            enhanced yellow fluorescent protein (luminopsin LMO2) for 
            bioluminescent optogenetics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7671)
  AUTHORS   Berglund K, Birkner E, Augustine GJ, Hochgeschwender U
  TITLE     Light-emitting channelrhodopsins for combined optogenetic and 
            chemical-genetic control of neurons.
  JOURNAL   PLoS One. 2013;8(3):e59759. doi: 10.1371/journal.pone.0059759. Epub 
            2013 Mar 27.
  PUBMED    23544095
REFERENCE   2  (bases 1 to 7671)
  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: "PLoS One.";
            date: "2013"; volume: "8(3):e59759. doi"; pages: " 
            10.1371/journal.pone.0059759. Epub 2013 Mar 27"
COMMENT     Sequence Label: pcDNA3.1-CMV-LMO2 (GLuc-VChR1-EYFP)
FEATURES             Location/Qualifiers
     source          1..7671
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      908..917
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             914..1468
                     /codon_start=1
                     /product="secreted Gaussia luciferase"
                     /label=hGLuc
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGKKLPLEVLKEMEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIVDIPEIPGFKDLEPMEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGGD"
     regulatory      1508..1517
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2423..2425,2426..3139)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2465..2486)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2726..2745)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3073..3094
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3169..3210
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3220..3237
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3260..3277)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3266..3490
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3536..3964
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3623..3642)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3833..3854
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3973..3993)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3978..4307
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4158..4293
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4220..4239
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4374..5168
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4428..4447)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5038..5057
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5344..5465
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5381..5400)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5435..5454
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5514..5530)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5514..5530)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5527..5549)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5538..5554
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5562..5568,5569..5586,5587..5592))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5607..5628
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5745..5762)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5916..6504)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5996..6015)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6675..7466,7467..7535))
                     /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     7298..7317
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7536..7640)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttgcc accatgggag tcaaagttct gtttgccctg atctgcatcg ctgtggccga
      961 ggccaagccc accgagaaca acgaagactt caacatcgtg gccgtggcca gcaacttcgc
     1021 gaccacggat ctcgatgctg accgcgggaa gttgcccggc aagaagctgc cgctggaggt
     1081 gctcaaagag atggaagcca atgcccggaa agctggctgc accaggggct gtctgatctg
     1141 cctgtcccac atcaagtgca cgcccaagat gaagaagttc atcccaggac gctgccacac
     1201 ctacgaaggc gacaaagagt ccgcacaggg cggcataggc gaggcgatcg tcgacattcc
     1261 tgagattcct gggttcaagg acttggagcc catggagcag ttcatcgcac aggtcgatct
     1321 gtgtgtggac tgcacaactg gctgcctcaa agggcttgcc aacgtgcagt gttctgacct
     1381 gctcaagaag tggctgccgc aacgctgtgc gacctttgcc agcaagatcc agggccaggt
     1441 ggacaagatc aagggggccg gtggtgacgg cgacccacta gtccagtgtg gtggaattgc
     1501 cggatccgcc accatggatt accctgttgc tagaagcctc atagttcgct acccaaccga
     1561 cctcggaaac ggcaccgtct gcatgccaag aggacagtgt tactgtgaag gttggcttcg
     1621 gagtcgcggc acttccattg aaaagacaat agcaattact cttcagtggg tagtctttgc
     1681 tttgtcagtg gcttgcctgg ggtggtatgc gtatcaagcg tggcgagcta cctgcggatg
     1741 ggaggaggtt tacgtagcct tgatagaaat gatgaaaagc atcatcgagg ccttccacga
     1801 gttcgacagc cctgcaacac tgtggctgtc ttcagggaac ggcgtagttt ggatgcggta
     1861 tggcgaatgg ctcctcacct gcccggtcct tctgatccat ctgagcaacc tcacaggcct
     1921 gaaggacgat tatagcaaaa ggactatggg cctgttggtt tctgatgtgg gatgcatcgt
     1981 gtggggcgca accagcgcca tgtgtacggg gtggacgaag atcctgttct tcctcatctc
     2041 attgagctat ggtatgtata cctattttca tgctgctaaa gtttatatcg aagcattcca
     2101 cacagttcca aaagggattt gtcgagaact ggtccgagtg atggcctgga cattctttgt
     2161 ggcttgggga atgtttccag tcctgtttct gctgggcacg gaaggattcg gtcatatcag
     2221 cccttatgga tctgccattg ggcactccat cctcgacctg attgcaaaga acatgtgggg
     2281 tgtgctgggg aattacctgc gcgtcaaaat ccacgagcac atcctgttgt atggcgacat
     2341 cagaaagaag cagaaaatta cgatcgccgg ccaagagatg gaggttgaga cactggtggc
     2401 tgaagaggag gacgcggccg ccgtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     2461 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     2521 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     2581 gcccgtgccc tggcccaccc tcgtgaccac cttcggctac ggcctgcagt gcttcgcccg
     2641 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     2701 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     2761 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     2821 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     2881 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     2941 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     3001 gctgctgccc gacaaccact acctgagcta ccagtccgcc ctgagcaaag accccaacga
     3061 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     3121 ggacgagctg tacaagtaac atatgctcta gagggcccgc ggttcgaagg taagcctatc
     3181 cctaaccctc tcctcggtct cgattctacg cgtaccggtc atcatcacca tcaccattga
     3241 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     3301 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     3361 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     3421 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     3481 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     3541 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3601 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3661 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3721 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3781 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3841 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3901 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3961 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     4021 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4081 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4141 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4201 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     4261 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     4321 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     4381 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4441 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4501 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     4561 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4621 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4681 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4741 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4801 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4861 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     4921 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4981 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5041 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5101 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5161 tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca
     5221 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     5281 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     5341 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     5401 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     5461 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     5521 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     5581 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     5641 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     5701 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5761 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5821 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5881 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5941 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     6001 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     6061 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     6121 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6181 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     6241 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6301 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     6361 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6421 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     6481 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6541 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6601 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6661 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6721 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6781 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6841 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6901 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6961 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     7021 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     7081 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     7141 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     7201 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     7261 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     7321 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     7381 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7441 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7501 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7561 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7621 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
