LOCUS       sequence-209658-        6688 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a chimeric G-protein coupled receptor 
            (Rhodopsin-GPR85; Opto-GPR85; D3).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6688)
  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 6688)
  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-GPR85
FEATURES             Location/Qualifiers
     source          1..6688
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..406
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        407..610
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     561..581
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     655..674
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        655..673
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             774..806
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     CDS             831..869
                     /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(2071..2088)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2077..2301
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2347..2775
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2434..2453)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2644..2665
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2784..2804)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2789..3118
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2969..3104
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3031..3050
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3185..3979
                     /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(3239..3258)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3849..3868
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4153..4274
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4190..4209)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4244..4263
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4323..4339)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4323..4339)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4336..4358)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4347..4363
                     /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(4371..4377,4378..4395,4396..4401))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4416..4437
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4554..4571)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4725..5313)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4805..4824)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5484..6275,6276..6344))
                     /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     6107..6126
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6345..6449)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6524..6543)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
       61 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      121 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      181 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      241 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      301 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      361 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      421 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      481 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      541 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      601 aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac
      661 gactcactat agggagacac aagctggcta gcgtttaaac gggccctcta gactcgagca
      721 ccatgaaaac gatcatcgcc ctgagctaca tcttctgcct ggtattcgcc atgtacaccg
      781 atatagagat gaacaggctg ggaaaggata gcctcatgaa cgggaccgag ggcccaaact
      841 tctacgtgcc tttctccaac aagacgggcg tggtgcgcag ccccttcgag gccccgcagt
      901 actacctggc ggagccatgg cagttctcca tgctggccgc ctacatgttc ctgctgatca
      961 tgcttggctt ccccatcaac ttcctcacgc tgtacgtcct agtgaaagat aagaccttgc
     1021 atagagcact caactacatc ctgctcaacc tggccgtggc cgacctcttc atggtgttcg
     1081 ggggcttcac caccaccctc tacacctctc tgcacgggta cttcgtcttt gggcccacgg
     1141 gctgcaacct ggagggcttc tttgccacct tgggcggtga aattgcactg tggtccttgg
     1201 tggtcctggc catcgagcgg tacgtggtgg tggcccatca ccgcttctat acaaagaggc
     1261 tgaccttttg gacggccatc atgggcgtcg ccttcacctg ggtcatggct ctggcctgtg
     1321 ccgcgccccc cctcgtcggc tggtccaggt acatcccgga gggcatgcag tgctcgtgcg
     1381 ggattgacta ctacacgccc cacgaggaaa ccaacaatga gtcgttcgtc atctacatgt
     1441 tcgtggtcca cttcatcatc cccctgattg tcatattctt ctgctacgga cacgatcgaa
     1501 gaaaaatgaa gccagtccag tttgtagcag cagtcagcca gaactggact tttcatggtc
     1561 ctggagccag tggccaggca gctgccaatt ggctagcagg atttggaagg ggtcccacac
     1621 cacccacctt gctgggcatc aggcaaaatg caaacaccac aggcagaaga aggctattgg
     1681 tcttagacga gttcaaaatg gagaaaagaa tcagccgcat ggtgatcatc atggtcatcg
     1741 ctttcctaat ctgctggctg ccctacgctg gggtggcgtt ctacatcttc acccatcagg
     1801 gctctgactt tggccccatc ttcatgacca tcccggcttt ctttgccaag acttctgccg
     1861 tctacaaccc cgtcatctac atcatgatga acaagcagtt ccgccgctgt ttcagcacaa
     1921 cccttcttta ctgcagaaaa tccaggttac caagggaacc ttactgtgtt ataacagaaa
     1981 ccagccaagt ggcgcctgcc taagaattcc accacactgg actagtggat ccgagctcgg
     2041 taccaagctt aagtttaaac cgctgatcag cctcgactgt gccttctagt tgccagccat
     2101 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     2161 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     2221 ggggtggggt ggggcaggac agcaaggggg aggattggga agagaatagc aggcatgctg
     2281 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     2341 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2401 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     2461 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     2521 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     2581 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     2641 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     2701 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     2761 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     2821 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     2881 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     2941 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3001 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     3061 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     3121 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     3181 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3241 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3301 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3361 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3421 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     3481 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     3541 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     3601 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     3661 cgaagaacat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     3721 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     3781 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     3841 ggacatagcg ttggctaccc gtgatattgc tgaagaactt ggcggcgaat gggctgaccg
     3901 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     3961 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4021 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4081 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4141 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4201 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4261 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     4321 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     4381 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4441 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4501 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttgcgc ttcctcgctc
     4561 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4621 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4681 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4741 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4801 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4861 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4921 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4981 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5041 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5101 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5161 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5221 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5281 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5341 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5401 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5461 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5521 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5581 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgggaccc acgctcaccg
     5641 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     5701 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     5761 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     5821 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     5881 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     5941 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6001 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6061 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6121 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6181 aggatcttac cactgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6241 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6301 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6361 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6421 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     6481 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
     6541 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     6601 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     6661 ttagggttag gcgttttgcg ctgcttcg
//
