LOCUS       sequence-396733-        6192 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 6192)
  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 6192)
  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-IRESWT-Gbetagamma ONE-GO
FEATURES             Location/Qualifiers
     source          1..6192
                     /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    1767..2340
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1790..2342
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1934..1951)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2161..2180
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2342..2362
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSK"
     CDS             2969..3481
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             3491..3544
                     /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             4565..4618
                     /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             4625..5359
                     /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     4762..4783
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4886..4907)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5147..5166)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    5604..6192
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5657..5677)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6075..6086)
                     /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 catgggctgc acgttgagcg ccgaggacaa
      721 ggcggcggtg gagcggagta agatgatcga ccgcaacttg cgggaggacg gagagaaagc
      781 ggccaaagaa gtgaagctgc tgctgctcgg cgctggagaa tctggtaaaa gtactattgt
      841 gaaacagatg aaaatcattc atgaggatgg ctattccgag gacgagtgta agcagtataa
      901 agttgtcgtc tacagcaata ccattcagtc catcattgca atcataagag ccatgggacg
      961 gttgaagatt gattttgggg aagctgccag agcggatgat gcccgacagt tatttgtttt
     1021 agctggcagt gctgaggaag gagtcatgac ttcagaacta gcaggcgtga ttaaacgttt
     1081 atggcgagat ggcggggtgc aggcatgctt cagcaggtcc agggaatatc agctcaatga
     1141 ttctgcttca tattacctaa atgatttgga tagaatatcc cagaccaact acattccaac
     1201 tcagcaagat gttcttcgga cgagagtgaa gactacaggc attgtggaga cccacttcac
     1261 cttcaaggag ctctacttca aaatgtttga tgtaggtggc caaagatccg aacggaaaaa
     1321 gtggatccac tgttttgagg gagtgacagc aattatcttt tgtgtggctc tcagtgatta
     1381 cgaccttgtt ctggctgagg acgaggaaat gaaccgaatg catgaaagca tgaaattgtt
     1441 tgacagcatt tgtaacaaca aatggtttac agacacttca atcattctct tccttaataa
     1501 gaaagacctt tttgaggaaa aaataaagag gagtccatta acaatctgtt atccagaata
     1561 cacaggttcc aatacgtacg aagaggcagc tgcttacatc cagtgccagt ttgaagatct
     1621 gaaccgaaga aaggacacca aggaggtcta cactcacttt acctgtgcca ctgacaccaa
     1681 aaacgtgcag tttgtttttg atgctgttac agatgtcatc attaaaaaca acttaaagga
     1741 atgtgggctt tactgagcgg ccgccgcccc tctccctccc ccccccctaa cgttactggc
     1801 cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg
     1861 ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct
     1921 aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca
     1981 gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg
     2041 aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct
     2101 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa
     2161 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt
     2221 atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa
     2281 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa
     2341 tatggggagt agcaagagca agacgtcgaa ttccggaatc aagcttcttg attgcgacca
     2401 agaactctac aagaacttcc ctctggtgat ctctgagcgc tggcagcagg aagtggcgga
     2461 aacagtttat gaagcagtaa atgcagacac ggataaaatc gaggccagga agagagctaa
     2521 aaataagcag cttggccacg aagaagatta cgccctggga agagactgca tcgtgcacgg
     2581 gtacatgctg aagctgggga accctttcct gacccagtgg cagcgccgct atttttacct
     2641 ctttccgaac agacttgagt ggagaggaga aggcgagtcg cgacaaagtt tactgacaat
     2701 ggaacagatt gtgtccgtgg aagaaactca gattaaagac aaaaagtgca ttttgttgag
     2761 aataaaagga gggaagcagt tcgttttgca gtgtgagagt gacccagagt ttgtgcagtg
     2821 gaagaaagag ctgacggaga cattcatgga ggcccagcgg ctgctacggc gagcccccaa
     2881 gttcctcaac aaatcccgct cagccgtcgt ggaactctca aagcctcccc tctgccatag
     2941 gaacagcaac ggcctcggag gtggatccat ggtcttcaca ctcgaagatt tcgttgggga
     3001 ctggcgacag acagccggct acaacctgga ccaagtcctt gaacagggag gtgtgtccag
     3061 tttgtttcag aatctcgggg tgtccgtaac tccgatccaa aggattgtcc tgagcggtga
     3121 aaatgggctg aagatcgaca tccatgtcat catcccgtat gaaggtctga gcggcgacca
     3181 aatgggccag atcgaaaaaa tttttaaggt ggtgtaccct gtggatgatc atcactttaa
     3241 ggtgatcctg cactatggca cactggtaat cgacggggtt acgccgaaca tgatcgacta
     3301 tttcggacgg ccgtatgaag gcatcgccgt gttcgacggc aaaaagatca ctgtaacagg
     3361 gaccctgtgg aacggcaaca aaattatcga cgagcgcctg atcaaccccg acggctccct
     3421 gctgttccga gtaaccatca acggagtgac cggctggcgg ctgtgcgaac gcattctggc
     3481 gggatcagga gaaggtagag gaagtctact aacatgcggg gacgtggagg aaaatcccgg
     3541 cccaatgggg gagcttgacc agttacggca ggaggccgag caacttaaga accagattcg
     3601 agacgccagg aaagcatgtg cagatgcaac tctctctcag atcacaaaca acatcgaccc
     3661 agtgggaaga atccaaatgc gcacgaggag gacactgcgg gggcacctgg ccaagatcta
     3721 cgccatgcac tggggcacag actccaggct tctcgtcagt gcctcgcagg atggtaaact
     3781 tatcatctgg gacagctaca ccaccaacaa ggtccacgcc atccctctgc gctcctcctg
     3841 ggtcatgacc tgtgcatatg ccccttctgg gaactatgtg gcctgcggtg gcctggataa
     3901 catttgctcc atttacaatc tgaaaactcg tgaggggaac gtgcgcgtga gtcgtgagct
     3961 ggcaggacac acaggttacc tgtcctgctg ccgattcctg gatgacaatc agatcgtcac
     4021 cagctctgga gacaccacgt gtgccctgtg ggacatcgag accggccagc agacgaccac
     4081 gtttaccgga cacactggag atgtcatgag cctttctctt gctcctgaca ccagactgtt
     4141 cgtctctggt gcttgtgatg cttcagccaa actctgggat gtgcgagaag gcatgtgccg
     4201 gcagaccttc actggccacg agtctgacat caatgccatt tgcttctttc caaatggcaa
     4261 tgcatttgcc actggctcag acgacgccac ctgcaggctg tttgaccttc gtgctgacca
     4321 ggagctcatg acttactccc atgacaacat catctgcggg atcacctctg tctccttctc
     4381 caagagcggg cgcctcctcc ttgctgggta cgacgacttc aactgcaacg tctgggatgc
     4441 actcaaagcc gaccgggcag gtgtcttggc tgggcatgac aaccgcgtca gctgcctggg
     4501 cgtgactgac gatggcatgg ctgtggcgac agggtcctgg gatagcttcc tcaagatctg
     4561 gaacgagggc agaggaagtc ttctaacatg cggtgacgtg gaggagaatc ccggccctac
     4621 cggtatggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     4681 cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccgccc tgagcaaaga
     4741 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     4801 tctcggcatg gacgagctgt acaagggtgg cagcggtggc atggtgagca agggcgagga
     4861 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     4921 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct
     4981 gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta
     5041 cggcctgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     5101 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     5161 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     5221 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     5281 cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa
     5341 gatccgccac aacatcgagt ccggactcag atctatggcc agcaacaaca ccgccagcat
     5401 agcacaagcc aggaagctgg tagagcagct taagatggaa gccaatatcg acaggataaa
     5461 ggtgtccaag gcagctgcag atttgatggc ctactgtgaa gcacatgcca aggaagaccc
     5521 cctcctgacc cctgttccgg cttcagaaaa cccgtttagg gagaagaagt ttttctgtgc
     5581 catcctttaa acgcgtctgg aacaatcaac ctctggatta caaaatttgt gaaagattga
     5641 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     5701 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     5761 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     5821 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     5881 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     5941 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc
     6001 tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     6061 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     6121 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     6181 ccgcctcccc gc
//
