LOCUS       sequence-209656-        6691 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a chimeric G-protein coupled receptor 
            (Rhodopsin-GPR83; Opto-GPR83; D1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6691)
  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 6691)
  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-GPR83
FEATURES             Location/Qualifiers
     source          1..6691
                     /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(2121..2138)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2127..2351
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2397..2825
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2484..2503)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2694..2715
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2834..2854)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2839..3168
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3019..3154
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3081..3100
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3235..4029
                     /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(3289..3308)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3899..3918
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4203..4324
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4240..4259)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4294..4313
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4373..4389)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4373..4389)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4386..4408)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4397..4413
                     /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(4421..4427,4428..4445,4446..4451))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4466..4487
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4604..4621)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4775..5363)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4855..4874)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5534..6325,6326..6394))
                     /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     6157..6176
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6395..6499)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6574..6593)
                     /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 acgtcatctt caagaaccag cgaatgcact cggccctcaa
     1081 ctacatcctg ctcaacctgg ccgtggccga cctcttcatg gtgttcgggg gcttcaccac
     1141 caccctctac acctctctgc acgggtactt cgtctttggg cccacgggct gcaacctgga
     1201 gggcttcttt gccaccttgg gcggtgaaat tgcactgtgg tccttggtgg tcctggccat
     1261 cgagcggtac gtggtggtga tgcacccctt gaaaccccgg atctcaatca caaaggccat
     1321 catgggcgtc gccttcacct gggtcatggc tctggcctgt gccgcgcccc ccctcgtcgg
     1381 ctggtccagg tacatcccgg agggcatgca gtgctcgtgc gggattgact actacacgcc
     1441 ccacgaggaa accaacaatg agtcgttcgt catctacatg ttcgtggtcc acttcatcat
     1501 ccccctgatt gtcatattct tctgctacgg acgtgtggcc aagaaactgt ggctgtgtaa
     1561 tatgattggc gatgtgacca cagagcagta ctttgccctg cggcgcaaaa agaagaaaac
     1621 catccgcatg gtgatcatca tggtcatcgc tttcctaatc tgctggctgc cctacgctgg
     1681 ggtggcgttc tacatcttca cccatcaggg ctctgacttt ggccccatct tcatgaccat
     1741 cccggctttc tttgccaaga cttctgccgt ctacaacccc gtcatctaca tcatgatgaa
     1801 caagcagttc cgcattgagc taaaggcatt actgagcatg tgtcaaagac ctcccaagcc
     1861 tcaggaggac aggccaccct ccccagttcc ttccttcagg gtggcctgga cagagaagaa
     1921 tgatggccag agggctcccc ttgccaataa cctcctgccc acctcccaac tccagtctgg
     1981 gaaaacagac ctgtcatctg tggaacccat tgtgacgatg agtacagaaa ccagccaagt
     2041 ggcgcctgcc taagaattcc accacactgg actagtggat ccgagctcgg taccaagctt
     2101 aagtttaaac cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     2161 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     2221 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     2281 ggggcaggac agcaaggggg aggattggga agagaatagc aggcatgctg gggatgcggt
     2341 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     2401 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2461 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2521 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     2581 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     2641 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2701 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     2761 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     2821 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     2881 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     2941 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3001 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3061 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     3121 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     3181 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3241 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3301 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3361 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     3421 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3481 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3541 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3601 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3661 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagaacat
     3721 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     3781 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     3841 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     3901 ttggctaccc gtgatattgc tgaagaactt ggcggcgaat gggctgaccg cttcctcgtg
     3961 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4021 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4081 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4141 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4201 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     4261 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     4321 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     4381 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     4441 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     4501 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     4561 gcgcggggag aggcggtttg cgtattgggc gctcttgcgc ttcctcgctc actgactcgc
     4621 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4681 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4741 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4801 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4861 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4921 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4981 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5041 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5101 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5161 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5221 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5281 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5341 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5401 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5461 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5521 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5581 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     5641 taccatctgg ccccagtgct gcaatgatac cgcgggaccc acgctcaccg gctccagatt
     5701 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5761 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5821 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5881 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5941 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6001 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6061 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6121 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6181 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6241 cactgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6301 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6361 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     6421 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6481 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     6541 gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     6601 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
     6661 ttaagctaca acaaggcaag gcttgaccga c
//
