LOCUS       sequence-396732-        6276 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 6276)
  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 6276)
  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-s-IRESWT-Gbetagamma ONE-GO
FEATURES             Location/Qualifiers
     source          1..6276
                     /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    1851..2424
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1874..2426
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2018..2035)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2245..2264
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2426..2446
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSK"
     CDS             3053..3565
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             3575..3628
                     /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             4649..4702
                     /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             4709..5443
                     /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     4846..4867
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4970..4991)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5231..5250)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    5688..6276
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5741..5761)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6159..6170)
                     /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 ggctgcctcg ggaacagtaa
      721 gaccgaggac cagcgcaacg aggagaaggc gcagcgtgag gccaacaaaa agatcgagaa
      781 gcagctgcag aaggacaagc aggtctaccg ggccacgcac cgcctgctgc tgctgggtgc
      841 tggagaatct ggtaaaagca ccattgtgaa gcagatgagg atcctgcatg ttaatgggtt
      901 taatggagac agtgagaagg caaccaaagt gcaggacatc aaaaacaacc tgaaagaggc
      961 gattgaaacc attgtggccg ccatgagcaa cctggtgccc cccgtggagc tggccaaccc
     1021 cgagaaccag ttcagagtgg actacattct gagtgtgatg aacgtgcctg actttgactt
     1081 ccctcccgaa ttctatgagc atgccaaggc tctgtgggag gatgaaggag tgcgtgcctg
     1141 ctacgaacgc tccaacgagt accagctgat tgactgtgcc cagtacttcc tggacaagat
     1201 cgacgtgatc aagcaggctg actatgtgcc gagcgatcag gacctgcttc gctgccgtgt
     1261 cctgacttct ggaatctttg agaccaagtt ccaggtggac aaagtcaact tccacatgtt
     1321 tgacgtgggt ggccagcgcg atgaacgccg caagtggatc cagtgcttca acgatgtgac
     1381 tgccatcatc ttcgtggtgg ccagcagcag ctacaacatg gtcatccggg aggacaacca
     1441 gaccaaccgc ctgcaggagg ctctgaacct cttcaagagc atctggaaca acagatggct
     1501 gcgcaccatc tctgtgatcc tgttcctcaa caagcaagat ctgctcgctg agaaagtcct
     1561 tgctgggaaa tcgaagattg aggactactt tccagaattt gctcgctaca ctactcctga
     1621 ggatgctact cccgagcccg gagaggaccc acgcgtgacc cgggccaagt acttcattcg
     1681 agatgagttt ctgaggatca gcactgccag tggagatggg cgtcactact gctaccctca
     1741 tttcacctgc gctgtggaca ctgagaacat ccgccgtgtg ttcaacgact gccgtgacat
     1801 cattcagcgc atgcaccttc gtcagtacga gctgctataa gcggccgccg cccctctccc
     1861 tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc
     1921 tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc
     1981 cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt
     2041 ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct
     2101 gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa
     2161 aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt
     2221 tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag
     2281 gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt
     2341 acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt
     2401 ttcctttgaa aaacacgatg ataatatggg gagtagcaag agcaagacgt cgaattccgg
     2461 aatcaagctt cttgattgcg accaagaact ctacaagaac ttccctctgg tgatctctga
     2521 gcgctggcag caggaagtgg cggaaacagt ttatgaagca gtaaatgcag acacggataa
     2581 aatcgaggcc aggaagagag ctaaaaataa gcagcttggc cacgaagaag attacgccct
     2641 gggaagagac tgcatcgtgc acgggtacat gctgaagctg gggaaccctt tcctgaccca
     2701 gtggcagcgc cgctattttt acctctttcc gaacagactt gagtggagag gagaaggcga
     2761 gtcgcgacaa agtttactga caatggaaca gattgtgtcc gtggaagaaa ctcagattaa
     2821 agacaaaaag tgcattttgt tgagaataaa aggagggaag cagttcgttt tgcagtgtga
     2881 gagtgaccca gagtttgtgc agtggaagaa agagctgacg gagacattca tggaggccca
     2941 gcggctgcta cggcgagccc ccaagttcct caacaaatcc cgctcagccg tcgtggaact
     3001 ctcaaagcct cccctctgcc ataggaacag caacggcctc ggaggtggat ccatggtctt
     3061 cacactcgaa gatttcgttg gggactggcg acagacagcc ggctacaacc tggaccaagt
     3121 ccttgaacag ggaggtgtgt ccagtttgtt tcagaatctc ggggtgtccg taactccgat
     3181 ccaaaggatt gtcctgagcg gtgaaaatgg gctgaagatc gacatccatg tcatcatccc
     3241 gtatgaaggt ctgagcggcg accaaatggg ccagatcgaa aaaattttta aggtggtgta
     3301 ccctgtggat gatcatcact ttaaggtgat cctgcactat ggcacactgg taatcgacgg
     3361 ggttacgccg aacatgatcg actatttcgg acggccgtat gaaggcatcg ccgtgttcga
     3421 cggcaaaaag atcactgtaa cagggaccct gtggaacggc aacaaaatta tcgacgagcg
     3481 cctgatcaac cccgacggct ccctgctgtt ccgagtaacc atcaacggag tgaccggctg
     3541 gcggctgtgc gaacgcattc tggcgggatc aggagaaggt agaggaagtc tactaacatg
     3601 cggggacgtg gaggaaaatc ccggcccaat gggggagctt gaccagttac ggcaggaggc
     3661 cgagcaactt aagaaccaga ttcgagacgc caggaaagca tgtgcagatg caactctctc
     3721 tcagatcaca aacaacatcg acccagtggg aagaatccaa atgcgcacga ggaggacact
     3781 gcgggggcac ctggccaaga tctacgccat gcactggggc acagactcca ggcttctcgt
     3841 cagtgcctcg caggatggta aacttatcat ctgggacagc tacaccacca acaaggtcca
     3901 cgccatccct ctgcgctcct cctgggtcat gacctgtgca tatgcccctt ctgggaacta
     3961 tgtggcctgc ggtggcctgg ataacatttg ctccatttac aatctgaaaa ctcgtgaggg
     4021 gaacgtgcgc gtgagtcgtg agctggcagg acacacaggt tacctgtcct gctgccgatt
     4081 cctggatgac aatcagatcg tcaccagctc tggagacacc acgtgtgccc tgtgggacat
     4141 cgagaccggc cagcagacga ccacgtttac cggacacact ggagatgtca tgagcctttc
     4201 tcttgctcct gacaccagac tgttcgtctc tggtgcttgt gatgcttcag ccaaactctg
     4261 ggatgtgcga gaaggcatgt gccggcagac cttcactggc cacgagtctg acatcaatgc
     4321 catttgcttc tttccaaatg gcaatgcatt tgccactggc tcagacgacg ccacctgcag
     4381 gctgtttgac cttcgtgctg accaggagct catgacttac tcccatgaca acatcatctg
     4441 cgggatcacc tctgtctcct tctccaagag cgggcgcctc ctccttgctg ggtacgacga
     4501 cttcaactgc aacgtctggg atgcactcaa agccgaccgg gcaggtgtct tggctgggca
     4561 tgacaaccgc gtcagctgcc tgggcgtgac tgacgatggc atggctgtgg cgacagggtc
     4621 ctgggatagc ttcctcaaga tctggaacga gggcagagga agtcttctaa catgcggtga
     4681 cgtggaggag aatcccggcc ctaccggtat ggacggcggc gtgcagctcg ccgaccacta
     4741 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     4801 ctaccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     4861 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg gtggcagcgg
     4921 tggcatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     4981 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     5041 ctacggcaag ctgaccctga agctgatctg caccaccggc aagctgcccg tgccctggcc
     5101 caccctcgtg accaccctgg gctacggcct gcagtgcttc gcccgctacc ccgaccacat
     5161 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     5221 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     5281 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     5341 gcacaagctg gagtacaact acaacagcca caacgtctat atcaccgccg acaagcagaa
     5401 gaacggcatc aaggccaact tcaagatccg ccacaacatc gagtccggac tcagatctat
     5461 ggccagcaac aacaccgcca gcatagcaca agccaggaag ctggtagagc agcttaagat
     5521 ggaagccaat atcgacagga taaaggtgtc caaggcagct gcagatttga tggcctactg
     5581 tgaagcacat gccaaggaag accccctcct gacccctgtt ccggcttcag aaaacccgtt
     5641 tagggagaag aagtttttct gtgccatcct ttaaacgcgt ctggaacaat caacctctgg
     5701 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     5761 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     5821 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     5881 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     5941 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     6001 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     6061 attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca
     6121 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     6181 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     6241 agacgagtcg gatctccctt tgggccgcct ccccgc
//
