LOCUS       sequence-209684-        7403 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a chimeric G-protein coupled receptor 
            (Rhodopsin-GPR153; Opto-GPR153; E7).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7403)
  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 7403)
  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-GPR153
FEATURES             Location/Qualifiers
     source          1..7403
                     /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(2833..2850)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2839..3063
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3109..3537
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3196..3215)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3406..3427
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3546..3566)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3551..3880
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3731..3866
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3947..4741
                     /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(4001..4020)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4611..4630
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4915..5036
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4952..4971)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5006..5025
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5085..5101)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5085..5101)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5098..5120)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5109..5125
                     /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(5133..5139,5140..5157,5158..5163))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5178..5199
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5316..5333)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5487..6075)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5567..5586)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6246..7037,7038..7106))
                     /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     6869..6888
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7107..7211)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7286..7305)
                     /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 gtacgtcgtt ggcgccaagc agaagaagtg gaagcccctc aactacatcc
     1081 tgctcaacct ggccgtggcc gacctcttca tggtgttcgg gggcttcacc accaccctct
     1141 acacctctct gcacgggtac ttcgtctttg ggcccacggg ctgcaacctg gagggcttct
     1201 ttgccacctt gggcggtgaa attgcactgt ggtccttggt ggtcctggcc atcgagcggt
     1261 acgtggtggt gtgctggcct gtcaactacc gattgagcaa tgccaagaag caggccatca
     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 cgctggccgt gcaggtgggg cgccaggccg
     1561 accgccgcgc cttcaccgtg cccaccatcg tggtggagga cgcgcagggc aagcggcgct
     1621 cctccatcga tggctcggag cccgccaaaa cctctctgca gcgcatggtg atcatcatgg
     1681 tcatcgcttt cctaatctgc tggctgccct acgctggggt ggcgttctac atcttcaccc
     1741 atcagggctc tgactttggc cccatcttca tgaccatccc ggctttcttt gccaagactt
     1801 ctgccgtcta caaccccgtc atctacatca tgatgaacaa gcagttccgc gctgacctca
     1861 aagctgtccg ggagaagtgc atggccctca tggccaacga cgaggagtca gacgatgaaa
     1921 ccagcctgga aggtggcatc tccccggacc tggtgttgga gcgctccctg gactatggct
     1981 atggaggtga ttttgtggcc ctagatagga tggccaagta tgagatctcc gccctggagg
     2041 ggggcctgcc ccagctctac ccactgcggc ccttgcagga ggacaagatg caatacctgc
     2101 aggtcccgcc cacgcggcgc ttctcccacg acgatgcgga cgtgtgggcc gccgtcccgc
     2161 tgcccgcctt cctgccgcgc tggggctccg gcgaggacct ggccgccctg gcgcacctgg
     2221 tgctgcctgc cgggcccgag cggcgccgcg ccagcctcct ggccttcgcg gaggacgcac
     2281 caccgtcccg cgcgcgccgc cgctcggccg agagcctgct gtcgctgcgg ccctcggccc
     2341 tggatagcgg cccgcgggga gcccgcgact cgccccccgg cagcccgcgc cgccgccccg
     2401 ggcccggccc ccgctccgcc tcggcctcgc tgctgcccga cgccttcgcc ctgaccgcct
     2461 tcgagtgcga gccacaggcc ctgcgccgcc cgcccgggcc cttccccgct gcgcccgccg
     2521 cccccgacgg cgcagatccc ggagaggccc cgacgccccc aagcagcgcc cagcggagcc
     2581 cagggccacg cccctctgcg cactcgcacg ccggctctct gcgccccggc ctgagcgcgt
     2641 cgtggggcga gcccgggggg ctgcgcgcgg cgggcggcgg cggcagcacc agcagcttcc
     2701 tgagttcccc ctccgagtcc tcgggctacg ccacgctgca ctcggactcg ctgggctccg
     2761 cgtccacaga aaccagccaa gtggcgcctg cctaactcga gtctagaggg cccgtttaaa
     2821 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     2881 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     2941 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     3001 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     3061 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     3121 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3181 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3241 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3301 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     3361 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3421 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     3481 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     3541 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     3601 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3661 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3721 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3781 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     3841 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     3901 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     3961 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4021 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4081 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     4141 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4201 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4261 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4321 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4381 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4441 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     4501 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4561 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4621 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4681 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4741 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     4801 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     4861 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     4921 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4981 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5041 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     5101 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     5161 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     5221 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5281 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5341 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5401 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5461 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5521 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5581 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5641 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5701 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5761 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5821 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5881 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5941 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6001 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6061 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6121 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6181 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6241 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6301 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6361 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6421 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6481 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6541 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6601 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6661 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6721 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6781 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6841 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6901 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6961 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7021 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7081 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7141 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7201 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc
     7261 ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     7321 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
     7381 caacaaggca aggcttgacc gac
//
