LOCUS       sequence-394364-        4336 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 4336)
  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 4336)
  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 ONE-GO
FEATURES             Location/Qualifiers
     source          1..4336
                     /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(2496..3070,3071..3073,3074..3082)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2663..2680)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2890..2909
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3093..3113
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSK"
     CDS             3219..3734
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     misc_feature    3748..4336
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3801..3821)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4219..4230)
                     /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 ggctgcacgt tgagcgccga
      721 ggacaaggcg gcggtggagc ggagtaagat gatcgaccgc aacttgcggg aggacggaga
      781 gaaagcggcc aaagaagtga agctgctgct gctcggcgct ggagaatctg gtaaaagtac
      841 tattgtgaaa cagatgaaaa tcattcatga ggatggctat tccgaggacg agtgtaagca
      901 gtataaagtt gtcgtctaca gcaataccat tcagtccatc attgcaatca taagagccat
      961 gggacggttg aagattgatt ttggggaagc tgccagagcg gatgatgccc gacagttatt
     1021 tgttttagct ggcagcgcta 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 gctgctgccc gacaaccact acctgagcta ccagtccgcc ctgagcaaag accccaacga
     1681 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     1741 ggacgagctg tacaagagcg ctgaggaagg agtcatgact tcagaactag caggcgtgat
     1801 taaacgttta tggcgagatg gcggggtgca ggcatgcttc agcaggtcca gggaatatca
     1861 gctcaatgat tctgcttcat attacctaaa tgatttggat agaatatccc agaccaacta
     1921 cattccaact cagcaagatg ttcttcggac gagagtgaag actacaggca ttgtggagac
     1981 ccacttcacc ttcaaggagc tctacttcaa aatgtttgat gtaggtggcc aaagatccga
     2041 acggaaaaag tggatccact gttttgaggg agtgacagca attatctttt gtgtggctct
     2101 cagtgattac gaccttgttc tggctgagga cgaggaaatg aaccgaatgc atgaaagcat
     2161 gaaattgttt gacagcattt gtaacaacaa atggtttaca gacacttcaa tcattctctt
     2221 ccttaataag aaagaccttt ttgaggaaaa aataaagagg agtccattaa caatctgtta
     2281 tccagaatac acaggttcca atacgtacga agaggcagct gcttacatcc agtgccagtt
     2341 tgaagatctg aaccgaagaa aggacaccaa ggaggtctac actcacttta cctgtgccac
     2401 tgacaccaaa aacgtgcagt ttgtttttga tgctgttaca gatgtcatca ttaaaaacaa
     2461 cttaaaggaa tgtgggcttt actgagcggc cgccgcccct ctccctcccc cccccctaac
     2521 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
     2581 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
     2641 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
     2701 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
     2761 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
     2821 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
     2881 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
     2941 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg
     3001 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
     3061 cgatgataag cttgccacaa cccgtaccaa agatggggag tagcaagagc aagacgtcga
     3121 attccggaat caagcttggt ggatcttctc gaggttacta ccatggtatt tgggtgggtg
     3181 aagaaggtcg actttctcga ggtggcaccg gtggatccat ggtcttcaca ctcgaagatt
     3241 tcgttgggga ctggcgacag acagccggct acaacctgga ccaagtcctt gaacagggag
     3301 gtgtgtccag tttgtttcag aatctcgggg tgtccgtaac tccgatccaa aggattgtcc
     3361 tgagcggtga aaatgggctg aagatcgaca tccatgtcat catcccgtat gaaggtctga
     3421 gcggcgacca aatgggccag atcgaaaaaa tttttaaggt ggtgtaccct gtggatgatc
     3481 atcactttaa ggtgatcctg cactatggca cactggtaat cgacggggtt acgccgaaca
     3541 tgatcgacta tttcggacgg ccgtatgaag gcatcgccgt gttcgacggc aaaaagatca
     3601 ctgtaacagg gaccctgtgg aacggcaaca aaattatcga cgagcgcctg atcaaccccg
     3661 acggctccct gctgttccga gtaaccatca acggagtgac cggctggcgg ctgtgcgaac
     3721 gcattctggc gtaaacgcgt ctggaacaat caacctctgg attacaaaat ttgtgaaaga
     3781 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     3841 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     3901 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     3961 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     4021 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     4081 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     4141 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     4201 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     4261 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     4321 tgggccgcct ccccgc
//
