LOCUS       sequence-396719-        5876 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 5876)
  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 5876)
  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-SV40-Galpha-s ONE-GO
FEATURES             Location/Qualifiers
     source          1..5876
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        50..366
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      217..352
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(708..710,711..1421)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(750..771)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1011..1030)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1358..1379
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(2290..2864,2865..2867,2868..2876)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2457..2474)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2684..2703
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2887..2907
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSK"
     CDS             3016..3528
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             3538..3591
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     misc_feature    5288..5876
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5341..5361)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5759..5770)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 caaaattttc gggtttatta cagggacagc agagatccag tttatcgatg gtgtggaaag
       61 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
      121 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
      181 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
      241 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
      301 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
      361 tgcaaactcg agactagttc tagagcggcc gcggatccat gggctgcctc gggaacagta
      421 agaccgagga ccagcgcaac gaggagaagg cgcagcgtga ggccaacaaa aagatcgaga
      481 agcagctgca gaaggacaag caggtctacc gggccacgca ccgcctgctg ctgctgggtg
      541 ctggagaatc tggtaaaagc accattgtga agcagatgag gatcctgcat gttaatgggt
      601 ttaatggaga cagtgagaag gcaaccaaag tgcaggacat caaaaacaac ctgaaagagg
      661 cgattgaaac cattgtggcc gccatgagca acctgggtgg cggaggggtg agcaagggcg
      721 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
      781 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
      841 agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg
      901 gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
      961 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     1021 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     1081 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     1141 acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact
     1201 tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga
     1261 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt
     1321 ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     1381 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gggcggtggg ggagtgcccc
     1441 ccgtggagct ggccaacccc gagaaccagt tcagagtgga ctacattctg agtgtgatga
     1501 acgtgcctga ctttgacttc cctcccgaat tctatgagca tgccaaggct ctgtgggagg
     1561 atgaaggagt gcgtgcctgc tacgaacgct ccaacgagta ccagctgatt gactgtgccc
     1621 agtacttcct ggacaagatc gacgtgatca agcaggctga ctatgtgccg agcgatcagg
     1681 acctgcttcg ctgccgtgtc ctgacttctg gaatctttga gaccaagttc caggtggaca
     1741 aagtcaactt ccacatgttt gacgtgggtg gccagcgcga tgaacgccgc aagtggatcc
     1801 agtgcttcaa cgatgtgact gccatcatct tcgtggtggc cagcagcagc tacaacatgg
     1861 tcatccggga ggacaaccag accaaccgcc tgcaggaggc tctgaacctc ttcaagagca
     1921 tctggaacaa cagatggctg cgcaccatct ctgtgatcct gttcctcaac aagcaagatc
     1981 tgctcgctga gaaagtcctt gctgggaaat cgaagattga ggactacttt ccagaatttg
     2041 ctcgctacac tactcctgag gatgctactc ccgagcccgg agaggaccca cgcgtgaccc
     2101 gggccaagta cttcattcga gatgagtttc tgaggatcag cactgccagt ggagatgggc
     2161 gtcactactg ctaccctcat ttcacctgcg ctgtggacac tgagaacatc cgccgtgtgt
     2221 tcaacgactg ccgtgacatc attcagcgca tgcaccttcg tcagtacgag ctgctataag
     2281 cggccgccgc ccctctccct cccccccccc taacgttact ggccgaagcc gcttggaata
     2341 aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt
     2401 gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct
     2461 cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc
     2521 ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga
     2581 caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc
     2641 ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt
     2701 attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg
     2761 gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc
     2821 gaaccacggg gacgtggttt tcctttgaaa aacacgatga taagcttgcc acaacccgta
     2881 ccaaagatgg ggagtagcaa gagcaagacg tcgaattccg gaatcaagct tggtggatca
     2941 tcaggaggta gcagggagct ggcctggggc atcagcgagt ggctggagga gtggggcagc
     3001 ggaggtggtg gatccatggt cttcacactc gaagatttcg ttggggactg gcgacagaca
     3061 gccggctaca acctggacca agtccttgaa cagggaggtg tgtccagttt gtttcagaat
     3121 ctcggggtgt ccgtaactcc gatccaaagg attgtcctga gcggtgaaaa tgggctgaag
     3181 atcgacatcc atgtcatcat cccgtatgaa ggtctgagcg gcgaccaaat gggccagatc
     3241 gaaaaaattt ttaaggtggt gtaccctgtg gatgatcatc actttaaggt gatcctgcac
     3301 tatggcacac tggtaatcga cggggttacg ccgaacatga tcgactattt cggacggccg
     3361 tatgaaggca tcgccgtgtt cgacggcaaa aagatcactg taacagggac cctgtggaac
     3421 ggcaacaaaa ttatcgacga gcgcctgatc aaccccgacg gctccctgct gttccgagta
     3481 accatcaacg gagtgaccgg ctggcggctg tgcgaacgca ttctggcggg atcaggagaa
     3541 ggtagaggaa gtctactaac atgcggggac gtggaggaaa atcccggccc aatggatgaa
     3601 gagcgcgccc tctacatcgt ccgggccggc gaggcggggg ctatcgagcg ggtcctgagg
     3661 gattacagcg acaagcatag ggctactttc aaatttgaat cagcagatga agataaaaga
     3721 aagaaactct gtgaaggcat atttaaggtc cttgtcaagg aggtcccaac gacatgccaa
     3781 gtgtcctgcc tggaagtact ccgcattctc tccagagaca aaaagatctt agttcctgta
     3841 acaactaagg aaaatatgca aatactgctg agactagcca agctccacga gtcagacgat
     3901 tctttggaga aggtgtcaga gttcccagtt attgtggaat cattgaagtg tctgtgtaac
     3961 atcgtgttca acagtcagat ggcacagcag cttagcctgg aactgaatct cgctgcaaag
     4021 ctctgtaacc tcctaaggaa gtgcaaggac cgaaaattta tcaatgacat aaagtgcttt
     4081 gacttgcgct tgctcttcgt cttgtcactt ttgcacactg acatcaggtc acaattgcgc
     4141 tatgagctcc agggactccc gctgctaacc cagatcttgg aaagtgcctt tagcatcaag
     4201 tggaccgatg agtatgagtc ggccatagac cataatggac ctcctctctc acctcaggag
     4261 acagactgtg ccattgaggc cctcaaagct ctgttcaatg tgacggtaga cagttggaag
     4321 gtgcataaag agagcgattc tcatcagttt cgtgtcatgg cagctgtcct tcgccattgt
     4381 ttactaattg taggtccaac tgaagacaaa accgaagagc tgcacagtaa tgccgtcaac
     4441 cttttaagca atgttccagt ctcttgtttg gatgttctca tttgtccatt aacccatgaa
     4501 gaaacagccc aagaggcagc aactctagat gaactgccca gtgataaaac aacggagaaa
     4561 gacacagctt tgaaaaacag taccatggta tacaatggca tgaatatgga ggccattcac
     4621 gttttactca attttatgga gaagagaatt gataagggaa gcagctatag agagggtcta
     4681 actccagttc tcagcttatt aacagaatgt tcccgagccc atcggaacat ccgaaaattt
     4741 ctcaaagatc aggttttacc gcctttgaga gatgtgacaa atcgaccaga agtcggctca
     4801 accgtgagaa ataagctggt tcgtctcatg acacatgttg acctcggagt caagcagatt
     4861 gctgctgagt tcctttttgt cctttgcaaa gagagagtgg atagcctgct gaaatacact
     4921 ggctatggga acgctgcagg actcttggcg gccaggggcc tcttggctgg aggaagagga
     4981 gataattggt actcagagga cgaggacacg gacactgaag aatacaaaaa cgcaaaacca
     5041 aacattaatc ttatcactgg tcatttagag gaaccaatgc caaaccctat agatgaaatg
     5101 acagaagaac aaaaagaata tgaagccatg aaacttgtca acatgcttga taaactttcc
     5161 agagaggagt tgcttagacc aatgggacta aaacctgacg ggacaataac gcctttggag
     5221 gaagcactca gccagtattc tgtcatcgaa gagaccagct cggacacaga ctgaacgcgt
     5281 ctggaacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     5341 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     5401 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     5461 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     5521 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     5581 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     5641 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     5701 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     5761 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     5821 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgc
//
