LOCUS       sequence-394356-        6175 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 6175)
  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 6175)
  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 ONE-GO
FEATURES             Location/Qualifiers
     source          1..6175
                     /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(1007..1009,1010..1720)
                     /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(1049..1070)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1310..1329)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1657..1678
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(2589..3163,3164..3166,3167..3175)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2756..2773)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2983..3002
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3186..3206
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSK"
     CDS             3315..3827
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             3837..3890
                     /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    5587..6175
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5640..5660)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6058..6069)
                     /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 cctgggtggc ggaggggtga gcaagggcga
     1021 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     1081 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     1141 gctgatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctggg
     1201 ctacggcctg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
     1261 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     1321 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     1381 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     1441 caacagccac aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt
     1501 caagatccgc cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa
     1561 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc
     1621 caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     1681 cgccgccggg atcactctcg gcatggacga gctgtacaag ggcggtgggg gagtgccccc
     1741 cgtggagctg gccaaccccg agaaccagtt cagagtggac tacattctga gtgtgatgaa
     1801 cgtgcctgac tttgacttcc ctcccgaatt ctatgagcat gccaaggctc tgtgggagga
     1861 tgaaggagtg cgtgcctgct acgaacgctc caacgagtac cagctgattg actgtgccca
     1921 gtacttcctg gacaagatcg acgtgatcaa gcaggctgac tatgtgccga gcgatcagga
     1981 cctgcttcgc tgccgtgtcc tgacttctgg aatctttgag accaagttcc aggtggacaa
     2041 agtcaacttc cacatgtttg acgtgggtgg ccagcgcgat gaacgccgca agtggatcca
     2101 gtgcttcaac gatgtgactg ccatcatctt cgtggtggcc agcagcagct acaacatggt
     2161 catccgggag gacaaccaga ccaaccgcct gcaggaggct ctgaacctct tcaagagcat
     2221 ctggaacaac agatggctgc gcaccatctc tgtgatcctg ttcctcaaca agcaagatct
     2281 gctcgctgag aaagtccttg ctgggaaatc gaagattgag gactactttc cagaatttgc
     2341 tcgctacact actcctgagg atgctactcc cgagcccgga gaggacccac gcgtgacccg
     2401 ggccaagtac ttcattcgag atgagtttct gaggatcagc actgccagtg gagatgggcg
     2461 tcactactgc taccctcatt tcacctgcgc tgtggacact gagaacatcc gccgtgtgtt
     2521 caacgactgc cgtgacatca ttcagcgcat gcaccttcgt cagtacgagc tgctataagc
     2581 ggccgccgcc cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa
     2641 ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg
     2701 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc
     2761 gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct
     2821 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac
     2881 aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc
     2941 cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta
     3001 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg
     3061 cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg
     3121 aaccacgggg acgtggtttt cctttgaaaa acacgatgat aagcttgcca caacccgtac
     3181 caaagatggg gagtagcaag agcaagacgt cgaattccgg aatcaagctt ggtggatcat
     3241 caggaggtag cagggagctg gcctggggca tcagcgagtg gctggaggag tggggcagcg
     3301 gaggtggtgg atccatggtc ttcacactcg aagatttcgt tggggactgg cgacagacag
     3361 ccggctacaa cctggaccaa gtccttgaac agggaggtgt gtccagtttg tttcagaatc
     3421 tcggggtgtc cgtaactccg atccaaagga ttgtcctgag cggtgaaaat gggctgaaga
     3481 tcgacatcca tgtcatcatc ccgtatgaag gtctgagcgg cgaccaaatg ggccagatcg
     3541 aaaaaatttt taaggtggtg taccctgtgg atgatcatca ctttaaggtg atcctgcact
     3601 atggcacact ggtaatcgac ggggttacgc cgaacatgat cgactatttc ggacggccgt
     3661 atgaaggcat cgccgtgttc gacggcaaaa agatcactgt aacagggacc ctgtggaacg
     3721 gcaacaaaat tatcgacgag cgcctgatca accccgacgg ctccctgctg ttccgagtaa
     3781 ccatcaacgg agtgaccggc tggcggctgt gcgaacgcat tctggcggga tcaggagaag
     3841 gtagaggaag tctactaaca tgcggggacg tggaggaaaa tcccggccca atggatgaag
     3901 agcgcgccct ctacatcgtc cgggccggcg aggcgggggc tatcgagcgg gtcctgaggg
     3961 attacagcga caagcatagg gctactttca aatttgaatc agcagatgaa gataaaagaa
     4021 agaaactctg tgaaggcata tttaaggtcc ttgtcaagga ggtcccaacg acatgccaag
     4081 tgtcctgcct ggaagtactc cgcattctct ccagagacaa aaagatctta gttcctgtaa
     4141 caactaagga aaatatgcaa atactgctga gactagccaa gctccacgag tcagacgatt
     4201 ctttggagaa ggtgtcagag ttcccagtta ttgtggaatc attgaagtgt ctgtgtaaca
     4261 tcgtgttcaa cagtcagatg gcacagcagc ttagcctgga actgaatctc gctgcaaagc
     4321 tctgtaacct cctaaggaag tgcaaggacc gaaaatttat caatgacata aagtgctttg
     4381 acttgcgctt gctcttcgtc ttgtcacttt tgcacactga catcaggtca caattgcgct
     4441 atgagctcca gggactcccg ctgctaaccc agatcttgga aagtgccttt agcatcaagt
     4501 ggaccgatga gtatgagtcg gccatagacc ataatggacc tcctctctca cctcaggaga
     4561 cagactgtgc cattgaggcc ctcaaagctc tgttcaatgt gacggtagac agttggaagg
     4621 tgcataaaga gagcgattct catcagtttc gtgtcatggc agctgtcctt cgccattgtt
     4681 tactaattgt aggtccaact gaagacaaaa ccgaagagct gcacagtaat gccgtcaacc
     4741 ttttaagcaa tgttccagtc tcttgtttgg atgttctcat ttgtccatta acccatgaag
     4801 aaacagccca agaggcagca actctagatg aactgcccag tgataaaaca acggagaaag
     4861 acacagcttt gaaaaacagt accatggtat acaatggcat gaatatggag gccattcacg
     4921 ttttactcaa ttttatggag aagagaattg ataagggaag cagctataga gagggtctaa
     4981 ctccagttct cagcttatta acagaatgtt cccgagccca tcggaacatc cgaaaatttc
     5041 tcaaagatca ggttttaccg cctttgagag atgtgacaaa tcgaccagaa gtcggctcaa
     5101 ccgtgagaaa taagctggtt cgtctcatga cacatgttga cctcggagtc aagcagattg
     5161 ctgctgagtt cctttttgtc ctttgcaaag agagagtgga tagcctgctg aaatacactg
     5221 gctatgggaa cgctgcagga ctcttggcgg ccaggggcct cttggctgga ggaagaggag
     5281 ataattggta ctcagaggac gaggacacgg acactgaaga atacaaaaac gcaaaaccaa
     5341 acattaatct tatcactggt catttagagg aaccaatgcc aaaccctata gatgaaatga
     5401 cagaagaaca aaaagaatat gaagccatga aacttgtcaa catgcttgat aaactttcca
     5461 gagaggagtt gcttagacca atgggactaa aacctgacgg gacaataacg cctttggagg
     5521 aagcactcag ccagtattct gtcatcgaag agaccagctc ggacacagac tgaacgcgtc
     5581 tggaacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     5641 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     5701 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     5761 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     5821 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     5881 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     5941 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg
     6001 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     6061 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     6121 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgc
//
