LOCUS       sequence-396731-        5119 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 5119)
  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 5119)
  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-IRESWT-Gbetagamma ONE-GO
FEATURES             Location/Qualifiers
     source          1..5119
                     /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"
     misc_feature    694..1267
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    717..1269
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(861..878)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1088..1107
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1269..1289
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSK"
     CDS             1896..2408
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             2418..2471
                     /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"
     CDS             3492..3545
                     /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"
     CDS             3552..4286
                     /codon_start=1
                     /product="circularly permuted form of the Venus yellow 
                     fluorescent protein (Nagai et al., 2004)"
                     /label=cp173Venus
                     /note="efficient FRET acceptor for CFP"
                     /translation="MDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRD
                     HMVLLEFVTAAGITLGMDELYKGGSGGMVSKGEELFTGVVPILVELDGDVNGHKFSVSG
                     EGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPE
                     GYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHN
                     VYITADKQKNGIKANFKIRHNIE"
     primer_bind     3689..3710
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3813..3834)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4074..4093)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    4531..5119
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4584..4604)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5002..5013)
                     /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 ccgcccctct ccctcccccc cccctaacgt
      721 tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac
      781 catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag
      841 cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa
      901 ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag
      961 gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga
     1021 tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag
     1081 agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc
     1141 ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag
     1201 gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg
     1261 atgataatat ggggagtagc aagagcaaga cgtcgaattc cggaatcaag cttcttgatt
     1321 gcgaccaaga actctacaag aacttccctc tggtgatctc tgagcgctgg cagcaggaag
     1381 tggcggaaac agtttatgaa gcagtaaatg cagacacgga taaaatcgag gccaggaaga
     1441 gagctaaaaa taagcagctt ggccacgaag aagattacgc cctgggaaga gactgcatcg
     1501 tgcacgggta catgctgaag ctggggaacc ctttcctgac ccagtggcag cgccgctatt
     1561 tttacctctt tccgaacaga cttgagtgga gaggagaagg cgagtcgcga caaagtttac
     1621 tgacaatgga acagattgtg tccgtggaag aaactcagat taaagacaaa aagtgcattt
     1681 tgttgagaat aaaaggaggg aagcagttcg ttttgcagtg tgagagtgac ccagagtttg
     1741 tgcagtggaa gaaagagctg acggagacat tcatggaggc ccagcggctg ctacggcgag
     1801 cccccaagtt cctcaacaaa tcccgctcag ccgtcgtgga actctcaaag cctcccctct
     1861 gccataggaa cagcaacggc ctcggaggtg gatccatggt cttcacactc gaagatttcg
     1921 ttggggactg gcgacagaca gccggctaca acctggacca agtccttgaa cagggaggtg
     1981 tgtccagttt gtttcagaat ctcggggtgt ccgtaactcc gatccaaagg attgtcctga
     2041 gcggtgaaaa tgggctgaag atcgacatcc atgtcatcat cccgtatgaa ggtctgagcg
     2101 gcgaccaaat gggccagatc gaaaaaattt ttaaggtggt gtaccctgtg gatgatcatc
     2161 actttaaggt gatcctgcac tatggcacac tggtaatcga cggggttacg ccgaacatga
     2221 tcgactattt cggacggccg tatgaaggca tcgccgtgtt cgacggcaaa aagatcactg
     2281 taacagggac cctgtggaac ggcaacaaaa ttatcgacga gcgcctgatc aaccccgacg
     2341 gctccctgct gttccgagta accatcaacg gagtgaccgg ctggcggctg tgcgaacgca
     2401 ttctggcggg atcaggagaa ggtagaggaa gtctactaac atgcggggac gtggaggaaa
     2461 atcccggccc aatgggggag cttgaccagt tacggcagga ggccgagcaa cttaagaacc
     2521 agattcgaga cgccaggaaa gcatgtgcag atgcaactct ctctcagatc acaaacaaca
     2581 tcgacccagt gggaagaatc caaatgcgca cgaggaggac actgcggggg cacctggcca
     2641 agatctacgc catgcactgg ggcacagact ccaggcttct cgtcagtgcc tcgcaggatg
     2701 gtaaacttat catctgggac agctacacca ccaacaaggt ccacgccatc cctctgcgct
     2761 cctcctgggt catgacctgt gcatatgccc cttctgggaa ctatgtggcc tgcggtggcc
     2821 tggataacat ttgctccatt tacaatctga aaactcgtga ggggaacgtg cgcgtgagtc
     2881 gtgagctggc aggacacaca ggttacctgt cctgctgccg attcctggat gacaatcaga
     2941 tcgtcaccag ctctggagac accacgtgtg ccctgtggga catcgagacc ggccagcaga
     3001 cgaccacgtt taccggacac actggagatg tcatgagcct ttctcttgct cctgacacca
     3061 gactgttcgt ctctggtgct tgtgatgctt cagccaaact ctgggatgtg cgagaaggca
     3121 tgtgccggca gaccttcact ggccacgagt ctgacatcaa tgccatttgc ttctttccaa
     3181 atggcaatgc atttgccact ggctcagacg acgccacctg caggctgttt gaccttcgtg
     3241 ctgaccagga gctcatgact tactcccatg acaacatcat ctgcgggatc acctctgtct
     3301 ccttctccaa gagcgggcgc ctcctccttg ctgggtacga cgacttcaac tgcaacgtct
     3361 gggatgcact caaagccgac cgggcaggtg tcttggctgg gcatgacaac cgcgtcagct
     3421 gcctgggcgt gactgacgat ggcatggctg tggcgacagg gtcctgggat agcttcctca
     3481 agatctggaa cgagggcaga ggaagtcttc taacatgcgg tgacgtggag gagaatcccg
     3541 gccctaccgg tatggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca
     3601 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag tccgccctga
     3661 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     3721 ggatcactct cggcatggac gagctgtaca agggtggcag cggtggcatg gtgagcaagg
     3781 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     3841 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     3901 tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     3961 tgggctacgg cctgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct
     4021 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     4081 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     4141 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     4201 actacaacag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca
     4261 acttcaagat ccgccacaac atcgagtccg gactcagatc tatggccagc aacaacaccg
     4321 ccagcatagc acaagccagg aagctggtag agcagcttaa gatggaagcc aatatcgaca
     4381 ggataaaggt gtccaaggca gctgcagatt tgatggccta ctgtgaagca catgccaagg
     4441 aagaccccct cctgacccct gttccggctt cagaaaaccc gtttagggag aagaagtttt
     4501 tctgtgccat cctttaaacg cgtctggaac aatcaacctc tggattacaa aatttgtgaa
     4561 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     4621 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     4681 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     4741 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     4801 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     4861 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     4921 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     4981 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     5041 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     5101 ctttgggccg cctccccgc
//
