LOCUS       sequence-396730-        3670 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  GPCR/G protein BRET biosensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3670)
  AUTHORS   Janicot R, Maziarz M, Park JC, Zhao J, Luebbers A, Green E, 
            Philibert CE, Zhang H, Layne MD, Wu JC, Garcia-Marcos M
  TITLE     Direct interrogation of context-dependent GPCR activity with a 
            universal biosensor platform.
  JOURNAL   Cell. 2024 Feb 16:S0092-8674(24)00065-5. doi: 
            10.1016/j.cell.2024.01.028.
  PUBMED    38412860
REFERENCE   2  (bases 1 to 3670)
  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: "Cell. 2024 
            Feb 16:S0092-8674(24)00065-5. doi: 10.1016/j.cell.2024.01.028."
COMMENT     Sequence Label: pLVX-CMV-Galpha-i3 G183S ONE-GO
FEATURES             Location/Qualifiers
     source          1..3670
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             join(374..376,377..379,380..1090)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(419..440)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(680..699)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1027..1048
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(1830..2404,2405..2407,2408..2416)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1997..2014)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2224..2243
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2427..2447
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSK"
     CDS             2553..3068
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     misc_feature    3082..3670
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3135..3155)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3553..3564)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 tcgagactag ttctagagcg gccgcggatc catgggctgc acgttgagcg ccgaggacaa
       61 ggcggcggtg gagcggagta agatgatcga ccgcaacttg cgggaggacg gagagaaagc
      121 ggccaaagaa gtgaagctgc tgctgctcgg cgctggagaa tctggtaaaa gtactattgt
      181 gaaacagatg aaaatcattc atgaggatgg ctattccgag gacgagtgta agcagtataa
      241 agttgtcgtc tacagcaata ccattcagtc catcattgca atcataagag ccatgggacg
      301 gttgaagatt gattttgggg aagctgccag agcggatgat gcccgacagt tatttgtttt
      361 agctggcagc gctatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
      421 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
      481 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
      541 gccctggccc accctcgtga ccaccttcgg ctacggcctg atgtgcttcg cccgctaccc
      601 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
      661 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
      721 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
      781 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
      841 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
      901 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
      961 gcccgacaac cactacctga gctaccagtc cgccctgagc aaagacccca acgagaagcg
     1021 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     1081 gctgtacaag agcgctgagg aaggagtcat gacttcagaa ctagcaggcg tgattaaacg
     1141 tttatggcga gatggcgggg tgcaggcatg cttcagcagg tccagggaat atcagctcaa
     1201 tgattctgct tcatattacc taaatgattt ggatagaata tcccagacca actacattcc
     1261 aactcagcaa gatgttcttc ggacgagagt gaagactaca agcattgtgg agacccactt
     1321 caccttcaag gagctctact tcaaaatgtt tgatgtaggt ggccaaagat ccgaacggaa
     1381 aaagtggatc cactgttttg agggagtgac agcaattatc ttttgtgtgg ctctcagtga
     1441 ttacgacctt gttctggctg aggacgagga aatgaaccga atgcatgaaa gcatgaaatt
     1501 gtttgacagc atttgtaaca acaaatggtt tacagacact tcaatcattc tcttccttaa
     1561 taagaaagac ctttttgagg aaaaaataaa gaggagtcca ttaacaatct gttatccaga
     1621 atacacaggt tccaatacgt acgaagaggc agctgcttac atccagtgcc agtttgaaga
     1681 tctgaaccga agaaaggaca ccaaggaggt ctacactcac tttacctgtg ccactgacac
     1741 caaaaacgtg cagtttgttt ttgatgctgt tacagatgtc atcattaaaa acaacttaaa
     1801 ggaatgtggg ctttactgag cggccgccgc ccctctccct cccccccccc taacgttact
     1861 ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata
     1921 ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt
     1981 cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa
     2041 gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag
     2101 cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca
     2161 cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc
     2221 aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat
     2281 tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta
     2341 aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga
     2401 taagcttgcc acaacccgta ccaaagatgg ggagtagcaa gagcaagacg tcgaattccg
     2461 gaatcaagct tggtggatct tctcgaggtt actaccatgg tatttgggtg ggtgaagaag
     2521 gtcgactttc tcgaggtggc accggtggat ccatggtctt cacactcgaa gatttcgttg
     2581 gggactggcg acagacagcc ggctacaacc tggaccaagt ccttgaacag ggaggtgtgt
     2641 ccagtttgtt tcagaatctc ggggtgtccg taactccgat ccaaaggatt gtcctgagcg
     2701 gtgaaaatgg gctgaagatc gacatccatg tcatcatccc gtatgaaggt ctgagcggcg
     2761 accaaatggg ccagatcgaa aaaattttta aggtggtgta ccctgtggat gatcatcact
     2821 ttaaggtgat cctgcactat ggcacactgg taatcgacgg ggttacgccg aacatgatcg
     2881 actatttcgg acggccgtat gaaggcatcg ccgtgttcga cggcaaaaag atcactgtaa
     2941 cagggaccct gtggaacggc aacaaaatta tcgacgagcg cctgatcaac cccgacggct
     3001 ccctgctgtt ccgagtaacc atcaacggag tgaccggctg gcggctgtgc gaacgcattc
     3061 tggcgtaaac gcgtctggaa caatcaacct ctggattaca aaatttgtga aagattgact
     3121 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     3181 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     3241 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     3301 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     3361 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     3421 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg
     3481 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     3541 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     3601 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     3661 gcctccccgc
//
