LOCUS       sequence-223843-        6733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a chimeric G-protein coupled receptor 
            (Rhodopsin-GPR62; Opto-GPR62; C6).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6733)
  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 6733)
  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-GPR62
FEATURES             Location/Qualifiers
     source          1..6733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             984..1016
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     CDS             1041..1079
                     /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(2326..2343)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2332..2556
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2602..3030
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2689..2708)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2899..2920
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3039..3059)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3044..3373
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3224..3359
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3286..3305
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3440..4234
                     /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(3494..3513)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4104..4123
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4408..4529
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4445..4464)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4499..4518
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4578..4594)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4578..4594)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4591..4613)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4602..4618
                     /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(4626..4632,4633..4650,4651..4656))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4671..4692
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4809..4826)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4980..5568)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5060..5079)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5739..6530,6531..6599))
                     /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     6362..6381
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6600..6704)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagacac aagctggcta
      901 gcgtttaaac gggccctcta gactcgagca ccatgaaaac gatcatcgcc ctgagctaca
      961 tcttctgcct ggtattcgcc atgtacaccg atatagagat gaacaggctg ggaaaggata
     1021 gcctcatgaa cgggaccgag ggcccaaact tctacgtgcc tttctccaac aagacgggcg
     1081 tggtgcgcag ccccttcgag gccccgcagt actacctggc ggagccatgg cagttctcca
     1141 tgctggccgc ctacatgttc ctgctgatca tgcttggctt ccccatcaac ttcctcacgc
     1201 tgtacgtcgt gctgcgcacg ccgggactgc gcgacctcaa ctacatcctg ctcaacctgg
     1261 ccgtggccga cctcttcatg gtgttcgggg gcttcaccac caccctctac acctctctgc
     1321 acgggtactt cgtctttggg cccacgggct gcaacctgga gggcttcttt gccaccttgg
     1381 gcggtgaaat tgcactgtgg tccttggtgg tcctggccat cgagcggtac gtggtggtgg
     1441 tgcacccgct gcggccaggc tcgcgcccgc cggccatcat gggcgtcgcc ttcacctggg
     1501 tcatggctct ggcctgtgcc gcgccccccc tcgtcggctg gtccaggtac atcccggagg
     1561 gcatgcagtg ctcgtgcggg attgactact acacgcccca cgaggaaacc aacaatgagt
     1621 cgttcgtcat ctacatgttc gtggtccact tcatcatccc cctgattgtc atattcttct
     1681 gctacggagg catcttcgtg gtggcgcgtc gcgctgccct gaggccccca cggccggcgc
     1741 gcgggtcccg actccactcg gactctctgg atagccgcct ttccatcttg ccgccgctcc
     1801 ggcctcgcct gcccgggggc aaggcggccc gcatggtgat catcatggtc atcgctttcc
     1861 taatctgctg gctgccctac gctggggtgg cgttctacat cttcacccat cagggctctg
     1921 actttggccc catcttcatg accatcccgg ctttctttgc caagacttct gccgtctaca
     1981 accccgtcat ctacatcatg atgaacaagc agttccgctt ggcactgggc cgcctctctc
     2041 gccgtgcact gcctggacct gtgcgggcct gcactccgca agcctggcac ccgcgggcac
     2101 tcttgcaatg cctccagagg cccccagagg gccctgccgt aggcccttct gaggctccag
     2161 aacagacccc cgagttggca ggagggcgga gccccgcata ccaggggcca cctgagagtt
     2221 ctctctccac agaaaccagc caagtggcgc ctgcctaaga attccaccac actggactag
     2281 tggatccgag ctcggtacca agcttaagtt taaaccgctg atcagcctcg actgtgcctt
     2341 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     2401 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     2461 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaga
     2521 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     2581 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     2641 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     2701 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     2761 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     2821 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     2881 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     2941 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     3001 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     3061 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3121 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3181 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3241 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3301 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3361 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     3421 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3481 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3541 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     3601 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     3661 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     3721 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     3781 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     3841 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     3901 tcaggatgat ctggacgaag aacatcaggg gctcgcgcca gccgaactgt tcgccaggct
     3961 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4021 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4081 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag aacttggcgg
     4141 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4201 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     4261 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     4321 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     4381 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     4441 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4501 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     4561 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     4621 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4681 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4741 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4801 tgcgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4861 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4921 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4981 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5041 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5101 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5161 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5221 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5281 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5341 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5401 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5461 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5521 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5581 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5641 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5701 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5761 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5821 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcgg
     5881 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5941 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6001 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6061 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6121 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6181 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6241 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6301 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6361 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6421 gggcgaaaac tctcaaggat cttaccactg ttgagatcca gttcgatgta acccactcgt
     6481 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6541 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6601 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6661 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6721 gtgccacctg acg
//
