LOCUS       sequence-209668-        6605 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a chimeric G-protein coupled receptor 
            (Rhodopsin-GPR139; Opto-GPR139; D10).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6605)
  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 6605)
  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-GPR139
FEATURES             Location/Qualifiers
     source          1..6605
                     /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(2035..2052)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2041..2265
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2311..2739
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2398..2417)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2608..2629
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2748..2768)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2753..3082
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2933..3068
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2995..3014
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3149..3943
                     /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(3203..3222)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3813..3832
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4117..4238
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4154..4173)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4208..4227
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4287..4303)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4287..4303)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4300..4322)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4311..4327
                     /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(4335..4341,4342..4359,4360..4365))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4380..4401
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4518..4535)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4689..5277)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4769..4788)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5448..6239,6240..6308))
                     /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     6071..6090
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6309..6413)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6488..6507)
                     /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 gtacgtcctc tcccagctgg tggcaagaag gcagaagctc aactacatcc
     1081 tgctcaacct ggccgtggcc gacctcttca tggtgttcgg gggcttcacc accaccctct
     1141 acacctctct gcacgggtac ttcgtctttg ggcccacggg ctgcaacctg gagggcttct
     1201 ttgccacctt gggcggtgaa attgcactgt ggtccttggt ggtcctggcc atcgagcggt
     1261 acgtggtggt gtgccacccg ctcaagtacc acacggtgtc atacccagcc cgcgccatca
     1321 tgggcgtcgc cttcacctgg gtcatggctc tggcctgtgc cgcgcccccc ctcgtcggct
     1381 ggtccaggta catcccggag ggcatgcagt gctcgtgcgg gattgactac tacacgcccc
     1441 acgaggaaac caacaatgag tcgttcgtca tctacatgtt cgtggtccac ttcatcatcc
     1501 ccctgattgt catattcttc tgctacggaa tcattgtgta caagctcagg aggaaaagca
     1561 attttcggct ccgtggctac tccacgggga aaaccacccg catggtgatc atcatggtca
     1621 tcgctttcct aatctgctgg ctgccctacg ctggggtggc gttctacatc ttcacccatc
     1681 agggctctga ctttggcccc atcttcatga ccatcccggc tttctttgcc aagacttctg
     1741 ccgtctacaa ccccgtcatc tacatcatga tgaacaagca gttccgcacc atggcagccg
     1801 ccacgctcaa ggctttcttc aagtgccaga agcaacctgt acagttctac accaatcata
     1861 acttttccat aacaagtagc ccctggatct cgccggcaaa ctcacactgc atcaagatgc
     1921 tggtgtacca gtatgacaaa aatggaaaac ctataaaagt atccccgaca gaaaccagcc
     1981 aagtggcgcc tgcctaactc gagtctagag ggcccgttta aacccgctga tcagcctcga
     2041 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     2101 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     2161 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     2221 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     2281 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     2341 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     2401 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     2461 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     2521 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     2581 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     2641 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     2701 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     2761 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2821 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2881 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     2941 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3001 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     3061 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     3121 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     3181 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3241 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3301 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     3361 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     3421 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     3481 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     3541 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     3601 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     3661 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     3721 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     3781 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     3841 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     3901 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     3961 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     4021 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4081 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     4141 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4201 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     4261 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     4321 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4381 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4441 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4501 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4561 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4621 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4681 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4741 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4801 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4861 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4921 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4981 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5041 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5101 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5161 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5221 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5281 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5341 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5401 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5461 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5521 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5581 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5641 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     5701 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5761 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5821 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5881 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5941 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6001 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6061 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6121 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6181 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6241 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6301 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6361 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6421 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
     6481 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     6541 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     6601 ccgac
//
