LOCUS       sequence-209699-        7066 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a chimeric G-protein coupled receptor 
            (Rhodopsin-GPR176; Opto-GPR176; F2).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7066)
  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 7066)
  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-GPR176
FEATURES             Location/Qualifiers
     source          1..7066
                     /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(2496..2513)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2502..2726
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2772..3200
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2859..2878)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3069..3090
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3209..3229)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3214..3543
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3394..3529
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3456..3475
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3610..4404
                     /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(3664..3683)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4274..4293
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4578..4699
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4615..4634)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4669..4688
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4748..4764)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4748..4764)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4761..4783)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4772..4788
                     /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(4796..4802,4803..4820,4821..4826))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4841..4862
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4979..4996)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5150..5738)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5230..5249)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5909..6700,6701..6769))
                     /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     6532..6551
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6770..6874)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6949..6968)
                     /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 acgtcacttg ccgcacaacc gtgttcaaat ctgtcctcaa
     1081 ctacatcctg ctcaacctgg ccgtggccga cctcttcatg gtgttcgggg gcttcaccac
     1141 caccctctac acctctctgc acgggtactt cgtctttggg cccacgggct gcaacctgga
     1201 gggcttcttt gccaccttgg gcggtgaaat tgcactgtgg tccttggtgg tcctggccat
     1261 cgagcggtac gtggtggtgc tctatccact ggagaggaaa atatctgatg ccaaggccat
     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 actgatccga cgggccctga gtgccagcca
     1561 gaagaagaag gtcatcatag cagcgctccg gaccccacag aacaccatct ctattcccta
     1621 tgcctcccag cgggaggccg agctgcaccg catggtgatc atcatggtca tcgctttcct
     1681 aatctgctgg ctgccctacg ctggggtggc gttctacatc ttcacccatc agggctctga
     1741 ctttggcccc atcttcatga ccatcccggc tttctttgcc aagacttctg ccgtctacaa
     1801 ccccgtcatc tacatcatga tgaacaagca gttccgcaag tgcttgatag ggaccctggt
     1861 gcaactacac caccgataca gtcgccgtaa tgtggtcagt acagggagtg gcatggctga
     1921 ggccagcctg gaacccagca tacgctcggg tagccagctc ctggagatgt tccacattgg
     1981 gcagcagcag atctttaagc ccacagagga tgaggaagag agtgaggcca agtacattgg
     2041 ttcagctgac ttccaggcca aggagatatt tagcacctgc ctggagggag agcaggggcc
     2101 acagtttgcg ccctctgccc cacccctgag cacagtggac tctgtatccc aggtggcacc
     2161 ggcagcccct gtggaacctg aaacattccc tgataagtat tccctgcagt ttggctttgg
     2221 gccttttgag ttgcctcctc agtggctctc agaaacccga aacagcaaga agcggctgct
     2281 tccccccttg ggcaacaccc cagaggagct gatccagaca aaggtgccca aggtaggcag
     2341 ggtggagcgg aagatgagca gaaacaataa agtgagcatt tttccaaagg tggattccac
     2401 agaaaccagc caagtggcgc ctgcctaaga attccaccac actggactag tggatccgag
     2461 ctcggtacca agcttaagtt taaaccgctg atcagcctcg actgtgcctt ctagttgcca
     2521 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     2581 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     2641 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaga atagcaggca
     2701 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     2761 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     2821 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     2881 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     2941 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3001 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3061 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3121 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     3181 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3241 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3301 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3361 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3421 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3481 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3541 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     3601 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3661 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3721 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3781 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3841 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3901 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     3961 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4021 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4081 ctggacgaag aacatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4141 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4201 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4261 tatcaggaca tagcgttggc tacccgtgat attgctgaag aacttggcgg cgaatgggct
     4321 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4381 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     4441 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4501 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4561 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4621 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4681 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     4741 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4801 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4861 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4921 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tgcgcttcct
     4981 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5041 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5101 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5161 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5221 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5281 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5341 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5401 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5461 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5521 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5581 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5641 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5701 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5761 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5821 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5881 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5941 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6001 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcgg gacccacgct
     6061 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6121 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6181 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6241 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6301 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6361 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6421 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6481 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6541 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6601 tctcaaggat cttaccactg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6661 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6721 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6781 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6841 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6901 acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc
     6961 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
     7021 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgac
//
