LOCUS       sequence-232387-        6614 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a chimeric G-protein coupled receptor 
            (Rhodopsin-GPR119; Opto-GPR119; D6).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6614)
  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 6614)
  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-GPR119
FEATURES             Location/Qualifiers
     source          1..6614
                     /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             976..1008
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     CDS             1033..1071
                     /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(2207..2224)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2213..2437
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2483..2911
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2570..2589)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2780..2801
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2920..2940)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2925..3254
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3105..3240
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3167..3186
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3321..4115
                     /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(3375..3394)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3985..4004
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4289..4410
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4326..4345)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4380..4399
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4459..4475)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4459..4475)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4472..4494)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4483..4499
                     /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(4507..4513,4514..4531,4532..4537))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4552..4573
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4690..4707)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4861..5449)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4941..4960)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5620..6411,6412..6480))
                     /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     6243..6262
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6481..6585)
                     /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 agggagaccc aagctggcta
      901 gcgtttaaac ttaagcttgc caccatgaaa acgatcatcg ccctgagcta catcttctgc
      961 ctggtattcg ccatgtacac cgatatagag atgaacaggc tgggaaagga tagcctcatg
     1021 aacgggaccg agggcccaaa cttctacgtg cctttctcca acaagacggg cgtggtgcgc
     1081 agccccttcg aggccccgca gtactacctg gcggagccat ggcagttctc catgctggcc
     1141 gcctacatgt tcctgctgat catgcttggc ttccccatca acttcctcac gctgtacgtc
     1201 gtgctgctgt tgatccacaa gaatgatggg ctcaactaca tcctgctcaa cctggccgtg
     1261 gccgacctct tcatggtgtt cgggggcttc accaccaccc tctacacctc tctgcacggg
     1321 tacttcgtct ttgggcccac gggctgcaac ctggagggct tctttgccac cttgggcggt
     1381 gaaattgcac tgtggtcctt ggtggtcctg gccatcgagc ggtacgtggt ggtgaagcag
     1441 cccttccgct acttgaagat catgagtggg ttcgtggcca tcatgggcgt cgccttcacc
     1501 tgggtcatgg ctctggcctg tgccgcgccc cccctcgtcg gctggtccag gtacatcccg
     1561 gagggcatgc agtgctcgtg cgggattgac tactacacgc cccacgagga aaccaacaat
     1621 gagtcgttcg tcatctacat gttcgtggtc cacttcatca tccccctgat tgtcatattc
     1681 ttctgctacg gagacatgct caagattgcc tccatgcaca gccagcagat tcgaaagatg
     1741 gaacatgcag gagccatggc tggaggttat cgatccccac ggactcccag cgacttcaaa
     1801 gctctccgca tggtgatcat catggtcatc gctttcctaa tctgctggct gccctacgct
     1861 ggggtggcgt tctacatctt cacccatcag ggctctgact ttggccccat cttcatgacc
     1921 atcccggctt tctttgccaa gacttctgcc gtctacaacc ccgtcatcta catcatgatg
     1981 aacaagcagt tccgcctgca gctctaccac atggccctag gagtgaagaa ggtgctcacc
     2041 tcattcctcc tctttctctc ggccaggaat tgtggcccag agaggcccag ggaaagttcc
     2101 tgtcacatcg tcactatctc cagctcagag tttgatggca cagaaaccag ccaagtggcg
     2161 cctgcctaac tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct
     2221 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     2281 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     2341 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     2401 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     2461 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2521 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2581 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2641 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     2701 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2761 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     2821 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     2881 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     2941 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3001 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3061 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3121 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3181 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3241 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     3301 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3361 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3421 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3481 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     3541 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     3601 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     3661 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     3721 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     3781 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     3841 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     3901 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     3961 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4021 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4081 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4141 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4201 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4261 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     4321 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4381 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     4441 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4501 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4561 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4621 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4681 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4741 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4801 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4861 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4921 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4981 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5041 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5101 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5161 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5221 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5281 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5341 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5401 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5461 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5521 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5581 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5641 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5701 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5761 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5821 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5881 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5941 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6001 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6061 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6121 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6181 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6241 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6301 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6361 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6421 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6481 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6541 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6601 agtgccacct gacg
//
