LOCUS       sequence-394360-        4372 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 4372)
  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 4372)
  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-z ONE-GO
FEATURES             Location/Qualifiers
     source          1..4372
                     /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(2532..3106,3107..3109,3110..3118)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2699..2716)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2926..2945
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3129..3149
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSK"
     CDS             3255..3770
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     misc_feature    3784..4372
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3837..3857)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4255..4266)
                     /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 ggctgtcggc agtcatcaga
      721 agaaaaggag gcggcacgac gatctaggcg catagatcgc catctgcgat ctgaatcaca
      781 gcggcaaagg cgcgaaatta agcttctctt gctgggaacc tccaattctg gcaaaagcac
      841 aatcgtaaaa caaatgaaga taattcattc aggcggtttt aatctcgagg cctgcaagga
      901 gtataaaccc ttgataatat ataacgccat agattcactc acacgaatta tccgggctct
      961 ggcagcgctt aaaattgact tccacaatcc tgaccgcgca tacgatgctg tacaactgtt
     1021 tgcattgacc ggcccggctg agtcaaaagg tgaacgatct ggaggtgggg gctctatggt
     1081 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     1141 cgtaaacggc cacaagttca gtgtgtccgg 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 agtctggcgg
     1801 tggaggctca attaccccgg agcttctggg agtcatgagg aggctctggg ccgaccccgg
     1861 agcacaagca tgcttcgggc ggagttcaga atatcacctg gaagataatg ctgcgtatta
     1921 tttgaatgac ttggaacgaa ttgctgctcc tgattatatc ccgaccgtgg aggacatttt
     1981 gcggtctagg gacatgacca cagggattgt cgagaacaaa ttcacgttta aggaacttac
     2041 attcaaaatg gtcgatgtgg gtggacagcg gtctgagcgg aaaaagtgga tacattgctt
     2101 cgagggggta acggccatta tcttttgtgt tgaactctct ggttacgatc tgaaattgta
     2161 cgaggacaac cagacgtccc gaatggccga aagtctccgc ttgtttgaca gcatttgcaa
     2221 caataactgg tttattaaca cttcactcat actttttctc aacaagaagg accttctgag
     2281 tgaaaaaata cgacgcattc cgcttagtgt gtgctttccg gagtacaagg ggcagaacac
     2341 ttatgaggag gctgccgtgt acattcagcg gcaattcgag gacctcaaca ggaataagga
     2401 gacaaaagaa atttactccc attttacttg tgcgaccgac acttccaata ttcagtttgt
     2461 ctttgacgct gtcaccgacg tcattatcca gaataacctt aagtacatag ggttgtgctg
     2521 agcggccgcc gcccctctcc ctcccccccc cctaacgtta ctggccgaag ccgcttggaa
     2581 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat
     2641 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct
     2701 ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct
     2761 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc
     2821 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa
     2881 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc
     2941 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg
     3001 gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc
     3061 ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataagcttg ccacaacccg
     3121 taccaaagat ggggagtagc aagagcaaga cgtcgaattc cggaatcaag cttggtggat
     3181 cttctcgagg ttactaccat ggtatttggg tgggtgaaga aggtcgactt tctcgaggtg
     3241 gcaccggtgg atccatggtc ttcacactcg aagatttcgt tggggactgg cgacagacag
     3301 ccggctacaa cctggaccaa gtccttgaac agggaggtgt gtccagtttg tttcagaatc
     3361 tcggggtgtc cgtaactccg atccaaagga ttgtcctgag cggtgaaaat gggctgaaga
     3421 tcgacatcca tgtcatcatc ccgtatgaag gtctgagcgg cgaccaaatg ggccagatcg
     3481 aaaaaatttt taaggtggtg taccctgtgg atgatcatca ctttaaggtg atcctgcact
     3541 atggcacact ggtaatcgac ggggttacgc cgaacatgat cgactatttc ggacggccgt
     3601 atgaaggcat cgccgtgttc gacggcaaaa agatcactgt aacagggacc ctgtggaacg
     3661 gcaacaaaat tatcgacgag cgcctgatca accccgacgg ctccctgctg ttccgagtaa
     3721 ccatcaacgg agtgaccggc tggcggctgt gcgaacgcat tctggcgtaa acgcgtctgg
     3781 aacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     3841 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     3901 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     3961 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     4021 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     4081 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     4141 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg
     4201 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     4261 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     4321 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gc
//
