LOCUS       sequence-265229-        6728 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a chimeric G-protein coupled receptor 
            (Rhodopsin-GPR135; Opto-GPR135; D9).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6728)
  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 6728)
  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-GPR135
FEATURES             Location/Qualifiers
     source          1..6728
                     /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(2158..2175)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2164..2388
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2434..2862
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2521..2540)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2731..2752
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2871..2891)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2876..3205
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3056..3191
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3118..3137
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3272..4066
                     /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(3326..3345)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3936..3955
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4240..4361
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4277..4296)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4331..4350
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4410..4426)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4410..4426)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4423..4445)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4434..4450
                     /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(4458..4464,4465..4482,4483..4488))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4503..4524
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4641..4658)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4812..5400)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4892..4911)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5571..6362,6363..6431))
                     /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     6194..6213
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6432..6536)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6611..6630)
                     /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 gtacgtcatt gtgaagcacc ggcagctccg caccgtcctc aactacatcc
     1081 tgctcaacct ggccgtggcc gacctcttca tggtgttcgg gggcttcacc accaccctct
     1141 acacctctct gcacgggtac ttcgtctttg ggcccacggg ctgcaacctg gagggcttct
     1201 ttgccacctt gggcggtgaa attgcactgt ggtccttggt ggtcctggcc atcgagcggt
     1261 acgtggtggt ggtgcgcccg ccgcgggaga agatcggccg ccgccgcgcc atcatgggcg
     1321 tcgccttcac ctgggtcatg gctctggcct gtgccgcgcc ccccctcgtc ggctggtcca
     1381 ggtacatccc ggagggcatg cagtgctcgt gcgggattga ctactacacg ccccacgagg
     1441 aaaccaacaa tgagtcgttc gtcatctaca tgttcgtggt ccacttcatc atccccctga
     1501 ttgtcatatt cttctgctac ggacacatct gcaagacggt gcgcctgtcg gacgtgcgcg
     1561 tgcggccggt gaacacctac gcgcgcgtgc tgcgcttctt cagcgaggtg cgcacgcgca
     1621 tggtgatcat catggtcatc gctttcctaa tctgctggct gccctacgct ggggtggcgt
     1681 tctacatctt cacccatcag ggctctgact ttggccccat cttcatgacc atcccggctt
     1741 tctttgccaa gacttctgcc gtctacaacc ccgtcatcta catcatgatg aacaagcagt
     1801 tccgccgcaa ccgcgaggag ggctaccgga ctaggaatgt ggacgctttc ctgcccagcc
     1861 agggcccggg tctgcaagcc agaagccgca gtcgccttcg aaaccgctat gccaaccggc
     1921 tgggggcctg caacaggatg tcctcttcca acccggccag cggagtggca ggggacgtgg
     1981 ccatgtgggc ccgcaaaaat ccagttgtac ttttctgccg agagggacca ccagagccgg
     2041 tgacggcagt gaccaaacag cctaaatccg aagctgggga taccagcctc acagaaacca
     2101 gccaagtggc gcctgcctaa ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct
     2161 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     2221 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     2281 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     2341 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     2401 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     2461 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     2521 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     2581 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     2641 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     2701 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     2761 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     2821 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     2881 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2941 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3001 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3061 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3121 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     3181 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     3241 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     3301 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     3361 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     3421 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     3481 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     3541 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     3601 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     3661 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     3721 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     3781 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     3841 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     3901 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     3961 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4021 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4081 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4141 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4201 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     4261 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4321 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     4381 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4441 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4501 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4561 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4621 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4681 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4741 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4801 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4861 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4921 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4981 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5041 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5101 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5161 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5221 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5281 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5341 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5401 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5461 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5521 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5581 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5641 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5701 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5761 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5821 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5881 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5941 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6001 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6061 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6121 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6181 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6241 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6301 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6361 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6421 tgaatactca tactctccct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6481 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6541 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     6601 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     6661 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     6721 tgaccgac
//
