LOCUS       sequence-209659-        6533 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a chimeric G-protein coupled receptor 
            (Rhodopsin-GPR87; Opto-GPR87; D4).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6533)
  AUTHORS   Morri M, Sanchez-Romero I, Tichy AM, Kainrath S, Gerrard EJ, 
            Hirschfeld PP, Schwarz J, Janovjak H
  TITLE     Optical functionalization of human Class A orphan G-protein-coupled 
            receptors.
  JOURNAL   Nat Commun. 2018 May 16;9(1):1950. doi: 10.1038/s41467-018-04342-1.
  PUBMED    29769519
REFERENCE   2  (bases 1 to 6533)
  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: "Nat 
            Commun."; date: "2018-05-16"; volume: "9(1):1950. doi"; pages: " 
            10.1038/s41467-018-04342-1"
COMMENT     Sequence Label: Opto-GPR87
FEATURES             Location/Qualifiers
     source          1..6533
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        74..453
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        454..657
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     608..628
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     702..721
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        702..720
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             813..845
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     CDS             870..908
                     /codon_start=1
                     /product="segment of bovine rhodopsin containing a signal 
                     for N-linked glycosylation (Favaloro et al., 2008)"
                     /label=opsin tag
                     /note="The asparagine in the NKT tripeptide will be 
                     glycosylated if this sequence is exposed to the ER lumen."
                     /translation="GPNFYVPFSNKTG"
     primer_bind     complement(1963..1980)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1969..2193
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2239..2667
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2326..2345)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2536..2557
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2676..2696)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2681..3010
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2861..2996
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2923..2942
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3077..3871
                     /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(3131..3150)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3741..3760
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4045..4166
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4082..4101)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4136..4155
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4215..4231)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4215..4231)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4228..4250)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4239..4255
                     /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(4263..4269,4270..4287,4288..4293))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4308..4329
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4446..4463)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4617..5205)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4697..4716)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5376..6167,6168..6236))
                     /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     5999..6018
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6237..6341)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6416..6435)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
       61 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
      121 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      181 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      241 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      301 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      361 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      421 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      481 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      541 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      601 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc
      661 tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg
      721 gagacccaag ctggctagcg tttaaactta agcttgccac catgaaaacg atcatcgccc
      781 tgagctacat cttctgcctg gtattcgcca tgtacaccga tatagagatg aacaggctgg
      841 gaaaggatag cctcatgaac gggaccgagg gcccaaactt ctacgtgcct ttctccaaca
      901 agacgggcgt ggtgcgcagc cccttcgagg ccccgcagta ctacctggcg gagccatggc
      961 agttctccat gctggccgcc tacatgttcc tgctgatcat gcttggcttc cccatcaact
     1021 tcctcacgct gtacgtcttt ttccacatta ggaataaaac cagcctcaac tacatcctgc
     1081 tcaacctggc cgtggccgac ctcttcatgg tgttcggggg cttcaccacc accctctaca
     1141 cctctctgca cgggtacttc gtctttgggc ccacgggctg caacctggag ggcttctttg
     1201 ccaccttggg cggtgaaatt gcactgtggt ccttggtggt cctggccatc gagcggtacg
     1261 tggtggtggt caagccattt ggggactctc ggatgtacag cataaccttc gccatcatgg
     1321 gcgtcgcctt cacctgggtc atggctctgg cctgtgccgc gccccccctc gtcggctggt
     1381 ccaggtacat cccggagggc atgcagtgct cgtgcgggat tgactactac acgccccacg
     1441 aggaaaccaa caatgagtcg ttcgtcatct acatgttcgt ggtccacttc atcatccccc
     1501 tgattgtcat attcttctgc tacggagcca tatccaggta catccacaaa tccagcaggc
     1561 aattcataag tcagtcaagc cgaaagcgaa aacataaccg catggtgatc atcatggtca
     1621 tcgctttcct aatctgctgg ctgccctacg ctggggtggc gttctacatc ttcacccatc
     1681 agggctctga ctttggcccc atcttcatga ccatcccggc tttctttgcc aagacttctg
     1741 ccgtctacaa ccccgtcatc tacatcatga tgaacaagca gttccgcaga aggctgttca
     1801 aaaaatcaaa tatcagaacc aggagtgaaa gcatcagatc actgcaaagt gtgagaagat
     1861 cggaagttcg catatattat gattacactg atgtgacaga aaccagccaa gtggcgcctg
     1921 cctaactcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta
     1981 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     2041 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     2101 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     2161 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     2221 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     2281 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     2341 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     2401 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     2461 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     2521 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     2581 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     2641 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     2701 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     2761 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2821 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     2881 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     2941 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3001 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     3061 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     3121 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     3181 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     3241 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     3301 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     3361 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     3421 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     3481 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     3541 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     3601 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     3661 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     3721 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     3781 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     3841 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     3901 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     3961 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     4021 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     4081 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     4141 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     4201 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     4261 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     4321 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     4381 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     4441 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4501 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4561 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4621 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4681 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4741 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4801 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4861 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4921 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4981 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5041 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5101 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5161 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5221 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5281 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5341 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5401 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     5461 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5521 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5581 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5641 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5701 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5761 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5821 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5881 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5941 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6001 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6061 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6121 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6181 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6241 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6301 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6361 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
     6421 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     6481 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gac
//
