LOCUS       sequence-394362-        4339 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 4339)
  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 4339)
  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-i2 ONE-GO
FEATURES             Location/Qualifiers
     source          1..4339
                     /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(1040..1042,1043..1045,1046..1756)
                     /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(1085..1106)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1346..1365)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1693..1714
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(2499..3073,3074..3076,3077..3085)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2666..2683)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2893..2912
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3096..3116
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSK"
     CDS             3222..3737
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     misc_feature    3751..4339
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3804..3824)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4222..4233)
                     /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 ggatgcaccg ttagcgccga
      721 agacaaagca gcggcggagc gaagcaagat gattgacaaa aatttgagag aggatggcga
      781 gaaggcggct cgcgaggtca aactgttgct tctgggtgcc ggagagagcg gcaagagcac
      841 tattgtaaaa cagatgaaga tcattcatga ggatggctac tcagaggagg aatgtcggca
      901 ataccgcgca gttgtgtata gtaatactat tcaatccatt atggcaattg taaaggcgat
      961 gggcaacctc cagatcgact ttgctgaccc gcagagggca gacgacgcga ggcagctgtt
     1021 cgccttgagc tgcgccgcta tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     1081 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     1141 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     1201 gcccgtgccc tggcccaccc tcgtgaccac cttcggctac ggcctgatgt gcttcgcccg
     1261 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     1321 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     1381 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     1441 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     1501 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     1561 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     1621 gttgctgccc gacaaccact acctgagcta ccagtccgcc ctgagcaaag accccaacga
     1681 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     1741 ggacgagctg tacaagagcg gcgaggaaca agggatgttg ccggaagatc tgtcaggagt
     1801 gattcggcgg ctgtgggccg accatggagt gcaagcgtgc tttgggagga gccgggaata
     1861 ccaactgaac gattccgccg cttattactt gaacgacctt gagcgcattg ctcaaagcga
     1921 ttatattcca actcaacaag atgtactgag aacacgcgtg aagacaacag gaatagtaga
     1981 gactcatttc acgtttaagg atctccactt taaaatgttc gatgttggag gacaacgatc
     2041 tgagcgcaag aagtggatac actgttttga gggcgttacc gcgatcattt tctgtgttgc
     2101 tttgtctgca tacgacctcg tcttggcaga ggatgaagaa atgaatcgaa tgcacgaatc
     2161 catgaagttg tttgattcta tatgcaacaa taagtggttt actgatacca gcatcatact
     2221 tttcttgaat aaaaaagact tgtttgagga gaagataaca cagagcccac ttacgatttg
     2281 tttcccggag tatacgggag cgaacaagta tgatgaagcc gcaagttata tccaatccaa
     2341 gtttgaggac ctgaataaga gaaaagacac aaaggagatt tacacacact tcacctgtgc
     2401 cactgacact aaaaacgtcc aatttgtttt cgacgccgtt accgatgtca taataaaaaa
     2461 taacctcaaa gactgtggtc ttttttaagc ggccgccgcc cctctccctc ccccccccct
     2521 aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt
     2581 tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg
     2641 acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc
     2701 gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt
     2761 tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta
     2821 taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg
     2881 gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag
     2941 gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag
     3001 tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa
     3061 acacgatgat aagcttgcca caacccgtac caaagatggg gagtagcaag agcaagacgt
     3121 cgaattccgg aatcaagctt ggtggatctt ctcgaggtta ctaccatggt atttgggtgg
     3181 gtgaagaagg tcgactttct cgaggtggca ccggtggatc catggtcttc acactcgaag
     3241 atttcgttgg ggactggcga cagacagccg gctacaacct ggaccaagtc cttgaacagg
     3301 gaggtgtgtc cagtttgttt cagaatctcg gggtgtccgt aactccgatc caaaggattg
     3361 tcctgagcgg tgaaaatggg ctgaagatcg acatccatgt catcatcccg tatgaaggtc
     3421 tgagcggcga ccaaatgggc cagatcgaaa aaatttttaa ggtggtgtac cctgtggatg
     3481 atcatcactt taaggtgatc ctgcactatg gcacactggt aatcgacggg gttacgccga
     3541 acatgatcga ctatttcgga cggccgtatg aaggcatcgc cgtgttcgac ggcaaaaaga
     3601 tcactgtaac agggaccctg tggaacggca acaaaattat cgacgagcgc ctgatcaacc
     3661 ccgacggctc cctgctgttc cgagtaacca tcaacggagt gaccggctgg cggctgtgcg
     3721 aacgcattct ggcgtaaacg cgtctggaac aatcaacctc tggattacaa aatttgtgaa
     3781 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     3841 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     3901 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     3961 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     4021 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     4081 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     4141 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     4201 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     4261 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     4321 ctttgggccg cctccccgc
//
