LOCUS       sequence-209688-        7357 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a chimeric G-protein coupled receptor 
            (Rhodopsin-GPR162; Opto-GPR162; E10).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7357)
  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 7357)
  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-GPR162
FEATURES             Location/Qualifiers
     source          1..7357
                     /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(2787..2804)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2793..3017
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3063..3491
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3150..3169)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3360..3381
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3500..3520)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3505..3834
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3685..3820
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3747..3766
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3901..4695
                     /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(3955..3974)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4565..4584
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4869..4990
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4906..4925)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4960..4979
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5039..5055)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5039..5055)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5052..5074)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5063..5079
                     /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(5087..5093,5094..5111,5112..5117))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5132..5153
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5270..5287)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5441..6029)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5521..5540)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6200..6991,6992..7060))
                     /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     6823..6842
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7061..7165)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7240..7259)
                     /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 acgtcatctc ggccaagcag cagaagcaca agccactcaa
     1081 ctacatcctg ctcaacctgg ccgtggccga cctcttcatg gtgttcgggg gcttcaccac
     1141 caccctctac acctctctgc acgggtactt cgtctttggg cccacgggct gcaacctgga
     1201 gggcttcttt gccaccttgg gcggtgaaat tgcactgtgg tccttggtgg tcctggccat
     1261 cgagcggtac gtggtggtgc gctggcccgt caactaccgc ctcagcaacg ccaagaagca
     1321 ggccatcatg ggcgtcgcct tcacctgggt catggctctg gcctgtgccg cgccccccct
     1381 cgtcggctgg tccaggtaca tcccggaggg catgcagtgc tcgtgcggga ttgactacta
     1441 cacgccccac gaggaaacca acaatgagtc gttcgtcatc tacatgttcg tggtccactt
     1501 catcatcccc ctgattgtca tattcttctg ctacggaaca ctgtgggccc ggccccggag
     1561 ggctcggcag gcccggagag tggggggtgg tggggggacc aaagcgggtg ggccaggggc
     1621 cttgggtacc cggccagctt ttgaggtacc agccattgtg gtggaggatg cccgagggaa
     1681 gcggcggtcc tcgctggatg gctcggagtc tgccaagaca tccctgcagc gcatggtgat
     1741 catcatggtc atcgctttcc taatctgctg gctgccctac gctggggtgg cgttctacat
     1801 cttcacccat cagggctctg actttggccc catcttcatg accatcccgg ctttctttgc
     1861 caagacttct gccgtctaca accccgtcat ctacatcatg atgaacaagc agttccgcgc
     1921 cgacgtgcgc acagtgtggg agcaatgcgt ggccatcatg tctgaggagg atggagatga
     1981 cgatgggggc tgtgacgact atgcagaggg ccgagtttgc aaagttcgct ttgatgctaa
     2041 cggagccaca ggaccaggga gccgggaccc cgcccaggtg aagctgctgc ctggaaggca
     2101 catgctgttc cctcctcttg agagagtcca ctacttacag gtccccctat cccggcgtct
     2161 gtcccatgat gagacaaaca tcttctctac ccctcgggaa ccaggctcct tcctgcacaa
     2221 gtggtcatcc tctgatgaca tccgggtcct cccagcccag agccgggccc tcgggggtcc
     2281 tcctgagtac ctgggacaaa gacacaggtt ggaggacgag gaggacgagg aagaggctga
     2341 aggtgggggg ctggccagcc ttcgccaatt cttggagagt ggggttctgg ggtcaggtgg
     2401 gggaccccca cggggtcctg gcttcttccg ggaggagatc accaccttca tcgatgagac
     2461 acctctgcct tctccgactg cctcaccagg gcactctcct cgtcggcccc ggccactggg
     2521 cctctcaccc cgccgactct cccttgggtc ccctgagagc agagccgttg gacttccttt
     2581 gggactaagc gcagggaggc gctgctccct gacggggggt gaagaaagtg caagggcttg
     2641 gggaggatcc tggggcccag gcaaccccat ctttccccag ctgaccctga cagaaaccag
     2701 ccaagtggcg cctgcctaag aattccacca cactggacta gtggatccga gctcggtacc
     2761 aagcttaagt ttaaaccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     2821 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2881 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     2941 tggggtgggg caggacagca agggggagga ttgggaagag aatagcaggc atgctgggga
     3001 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     3061 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3121 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3181 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     3241 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     3301 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3361 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3421 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3481 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3541 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3601 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3661 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3721 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     3781 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     3841 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     3901 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     3961 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4021 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4081 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4141 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4201 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4261 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4321 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4381 gaacatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     4441 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4501 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4561 atagcgttgg ctacccgtga tattgctgaa gaacttggcg gcgaatgggc tgaccgcttc
     4621 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4681 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     4741 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4801 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4861 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4921 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4981 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     5041 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5101 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5161 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5221 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttgcgcttcc tcgctcactg
     5281 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5341 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5401 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5461 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5521 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5581 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5641 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5701 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5761 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5821 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5881 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5941 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6001 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6061 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6121 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6181 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6241 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6301 agggcttacc atctggcccc agtgctgcaa tgataccgcg ggacccacgc tcaccggctc
     6361 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6421 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6481 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6541 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6601 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6661 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6721 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6781 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6841 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6901 tcttaccact gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6961 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7021 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7081 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7141 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     7201 gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc
     7261 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
     7321 caaaatttaa gctacaacaa ggcaaggctt gaccgac
//
