LOCUS       sequence-394357-        4369 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 4369)
  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 4369)
  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-oA ONE-GO
FEATURES             Location/Qualifiers
     source          1..4369
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     enhancer        58..361
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        362..565
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     516..536
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     562..586
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(1076..1078,1079..1081,1082..1792)
                     /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(1121..1142)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1382..1401)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1729..1750
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(2529..3103,3104..3106,3107..3115)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2696..2713)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2923..2942
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3126..3146
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSK"
     CDS             3252..3767
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     misc_feature    3781..4369
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3834..3854)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4252..4263)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 tcgggtttat tacagggaca gcagagatcc agtttatcga taagcttggg agttccgcgt
       61 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      121 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      181 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      241 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      301 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      361 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
      421 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      481 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
      541 gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagacgcca
      601 tccacgctgt tttgacctcc atagaagaca ccgactctac tagaggatct atttccggtg
      661 aattcctcga gactagttct agagcggccg cggatccatg ggatgtactc tgagcgcaga
      721 ggagagagcc gccctcgagc ggagcaaggc gattgagaaa aatctcaaag aagatggcat
      781 cagcgccgcc aaagacgtga aattactcct gctgggggct ggagaatcag gaaaaagcac
      841 cattgtgaag cagatgaaga tcatccatga agatggcttc tctggagaag acgtaaagca
      901 gtacaagcct gtcgtctaca gcaacaccat ccagtctctg gcagccattg tgcgggccat
      961 ggatactctg ggcgtggagt atggtgacaa ggagaggaag gcagactcca agatggtgtg
     1021 tgacgtggtg agtcgcatgg aggacactga accaagatcc ggtgggggcg ggagcatggt
     1081 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     1141 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     1201 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     1261 gaccaccttc ggctacggcc tgatgtgctt cgcccgctac cccgaccaca tgaagcagca
     1321 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1381 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1441 ccgcattgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     1501 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     1561 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     1621 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     1681 gagctaccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     1741 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtccggagg
     1801 tggaggatct ttctctgcag aactgctttc tgccatgatg cgactctggg gcgactcggg
     1861 gatccaggag tgcttcaacc gatctcggga gtatcagctc aacgactctg ccaaatacta
     1921 cctggacagc ttggatcgga ttggagctgc agaatacatg cccaccgagc aggacatcct
     1981 ccgaaccagg gtcaaaacaa ctggcatcgt agaaacccac ttcaccttca agaacctcca
     2041 cttcaggctg tttgacgttg ggggccagcg atctgaacgt aagaagtgga tccactgctt
     2101 cgaggatgtc acggccatca tcttctgtgt cgcactcagc ggctatgacc aggtgctcca
     2161 cgaggacgaa accacgaacc gcatgcacga gtctctcatg ctcttcgact ccatctgtaa
     2221 caacaagttt ttcatcgata cctccatcat tctcttcctc aacaagaaag acctctttgg
     2281 cgagaagatt aagaagtcac ccttgaccat ctgctttcct gaatacccag gctccaacac
     2341 ctatgaagac gcagctgcct acatccaaac acagtttgaa agcaaaaacc gctcacccaa
     2401 caaagaaatt tactgtcaca tgacttgtgc cacagacacg aataatatcc aggtggtatt
     2461 cgacgccgtc accgacatca tcattgccaa caatctccgg ggctgtggct tgtactgagc
     2521 ggccgccgcc cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa
     2581 ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg
     2641 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc
     2701 gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct
     2761 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac
     2821 aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc
     2881 cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta
     2941 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg
     3001 cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg
     3061 aaccacgggg acgtggtttt cctttgaaaa acacgatgat aagcttgcca caacccgtac
     3121 caaagatggg gagtagcaag agcaagacgt cgaattccgg aatcaagctt ggtggatctt
     3181 ctcgaggtta ctaccatggt atttgggtgg gtgaagaagg tcgactttct cgaggtggca
     3241 ccggtggatc catggtcttc acactcgaag atttcgttgg ggactggcga cagacagccg
     3301 gctacaacct ggaccaagtc cttgaacagg gaggtgtgtc cagtttgttt cagaatctcg
     3361 gggtgtccgt aactccgatc caaaggattg tcctgagcgg tgaaaatggg ctgaagatcg
     3421 acatccatgt catcatcccg tatgaaggtc tgagcggcga ccaaatgggc cagatcgaaa
     3481 aaatttttaa ggtggtgtac cctgtggatg atcatcactt taaggtgatc ctgcactatg
     3541 gcacactggt aatcgacggg gttacgccga acatgatcga ctatttcgga cggccgtatg
     3601 aaggcatcgc cgtgttcgac ggcaaaaaga tcactgtaac agggaccctg tggaacggca
     3661 acaaaattat cgacgagcgc ctgatcaacc ccgacggctc cctgctgttc cgagtaacca
     3721 tcaacggagt gaccggctgg cggctgtgcg aacgcattct ggcgtaaacg cgtctggaac
     3781 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     3841 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     3901 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     3961 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     4021 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     4081 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     4141 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc
     4201 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     4261 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     4321 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgc
//
