LOCUS       sequence-209637-        7420 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a chimeric G-protein coupled receptor 
            (Rhodopsin-GPR50; Opto-GPR50; C2).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7420)
  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 7420)
  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-GPR50
FEATURES             Location/Qualifiers
     source          1..7420
                     /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             821..853
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     CDS             878..916
                     /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(2850..2867)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2856..3080
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3126..3554
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3213..3232)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3423..3444
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3563..3583)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3568..3897
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3748..3883
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3810..3829
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3964..4758
                     /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(4018..4037)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4628..4647
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4932..5053
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4969..4988)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5023..5042
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5102..5118)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5102..5118)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5115..5137)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5126..5142
                     /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(5150..5156,5157..5174,5175..5180))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5195..5216
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5333..5350)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5504..6092)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5584..5603)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6263..7054,7055..7123))
                     /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     6886..6905
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7124..7228)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7303..7322)
                     /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 gagacacaag ctggctagcg tttaaacggg ccctctagac tcgagcacca tgaaaacgat
      781 catcgccctg agctacatct tctgcctggt attcgccatg tacaccgata tagagatgaa
      841 caggctggga aaggatagcc tcatgaacgg gaccgagggc ccaaacttct acgtgccttt
      901 ctccaacaag acgggcgtgg tgcgcagccc cttcgaggcc ccgcagtact acctggcgga
      961 gccatggcag ttctccatgc tggccgccta catgttcctg ctgatcatgc ttggcttccc
     1021 catcaacttc ctcacgctgt acgtcgtgac gaagaacaag aagctccgga attctctcaa
     1081 ctacatcctg ctcaacctgg ccgtggccga cctcttcatg gtgttcgggg gcttcaccac
     1141 caccctctac acctctctgc acgggtactt cgtctttggg cccacgggct gcaacctgga
     1201 gggcttcttt gccaccttgg gcggtgaaat tgcactgtgg tccttggtgg tcctggccat
     1261 cgagcggtac gtggtggtgt gccacagcct ccagtacgaa cggatcttca gtgtgcgcaa
     1321 tgccatcatg ggcgtcgcct tcacctgggt catggctctg gcctgtgccg cgccccccct
     1381 cgtcggctgg tccaggtaca tcccggaggg catgcagtgc tcgtgcggga ttgactacta
     1441 cacgccccac gaggaaacca acaatgagtc gttcgtcatc tacatgttcg tggtccactt
     1501 catcatcccc ctgattgtca tattcttctg ctacggaagg atctggacca aagtgctggc
     1561 ggcccgtgac cctgcagggc agaatcctga caaccaactt gctgaggttc gcaattttcg
     1621 catggtgatc atcatggtca tcgctttcct aatctgctgg ctgccctacg ctggggtggc
     1681 gttctacatc ttcacccatc agggctctga ctttggcccc atcttcatga ccatcccggc
     1741 tttctttgcc aagacttctg ccgtctacaa ccccgtcatc tacatcatga tgaacaagca
     1801 gttccgcaga gaatactgga ccatcttcca tgctatgcgg caccctatca tattcttctc
     1861 tggcctcatc agtgatattc gtgagatgca ggaggcccgt accctggccc gcgcccgtgc
     1921 ccatgctcgc gaccaagctc gtgaacaaga ccgtgcccat gcctgtcctg ctgtggagga
     1981 aaccccgatg aatgtccgga atgttccatt acctggtgat gctgcagctg gccaccccga
     2041 ccgtgcctct ggccacccta agccccattc cagatcctcc tctgcctatc gcaaatctgc
     2101 ctctacccac cacaagtctg tctttagcca ctccaaggct gcctctggtc acctcaagcc
     2161 tgtctctggc cactccaagc ctgcctctgg tcaccccaag tctgccactg tctaccctaa
     2221 gcctgcctct gtccatttca aggctgactc tgtccatttc aagggtgact ctgtccattt
     2281 caagcctgac tctgttcatt tcaagcctgc ttccagcaac cccaagccca tcactggcca
     2341 ccatgtctct gctggcagcc actccaagtc tgccttcagt gctgccacca gccaccctaa
     2401 acccaccact ggccacatca agccagctac cagccatgca gagcccacca ctgctgacta
     2461 tcccaagcct gccactacca gccaccctaa gcccactgct gctgacaacc ctgagctctc
     2521 tgcctcccat tgccccgaga tccctgccat tgcccaccct gtgtctgacg acagtgacct
     2581 ccctgagtcg gcctctagcc ctgccgctgg gcccaccaag cctgctgcca gccagctgga
     2641 gtctgacacc atcgctgacc ttcctgaccc tactgtagtc actaccagta ccaatgatta
     2701 ccatgatgtc gtggttattg atgttgaaga tgatcctgat gaaatggctg tgacagaaac
     2761 cagccaagtg gcgcctgcct aagaattcca ccacactgga ctagtggatc cgagctcggt
     2821 accaagctta agtttaaacc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     2881 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     2941 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     3001 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gagaatagca ggcatgctgg
     3061 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     3121 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3181 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3241 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3301 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3361 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3421 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     3481 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3541 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3601 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3661 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3721 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3781 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     3841 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     3901 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     3961 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4021 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4081 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4141 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4201 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4261 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4321 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4381 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4441 gaagaacatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     4501 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4561 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4621 gacatagcgt tggctacccg tgatattgct gaagaacttg gcggcgaatg ggctgaccgc
     4681 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4741 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4801 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4861 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4921 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     4981 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5041 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     5101 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5161 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5221 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5281 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttgcgct tcctcgctca
     5341 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5401 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5461 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5521 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5581 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5641 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5701 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5761 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5821 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5881 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5941 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6001 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     6061 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6121 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6181 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6241 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6301 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6361 gggagggctt accatctggc cccagtgctg caatgatacc gcgggaccca cgctcaccgg
     6421 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6481 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6541 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6601 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6661 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6721 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6781 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6841 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6901 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6961 ggatcttacc actgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7021 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7081 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7141 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7201 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     7261 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
     7321 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
     7381 gagcaaaatt taagctacaa caaggcaagg cttgaccgac
//
