LOCUS       sequence-394366-        4981 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 4981)
  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 4981)
  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-12 ONE-GO
FEATURES             Location/Qualifiers
     source          1..4981
                     /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"
     primer_bind     complement(1433..1452)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             1592..1843
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNH
                     YLSYQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     1780..1801
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(2598..3172,3173..3175,3176..3184)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2765..2782)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2992..3011
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3195..3215
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSK"
     CDS             3864..4379
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     misc_feature    4393..4981
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4446..4466)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4864..4875)
                     /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 tcaggtgtag taaggacatt
      721 gtcccgatgt ctgttgcccg cagaggcggg gggcgctcgc gaaagacgcg cgggctctgg
      781 cgcccgggac gctgaaaggg aagcgcgaag aagatcacgc gatattgacg cgcttcttgc
      841 gcgggaaagg agagcggtcc gcagactcgt gaaaatattg ctgctcgggg ctggcgagtc
      901 tggcaaatcc acgttcctca aacaaatgag aatcatacac gggcgggaat tcgaccaaaa
      961 agctttgctc gaattcaggg acactatctt tgataatata ctcaaagggt cccgggttct
     1021 cgttgatgcc agggacaagc tcggcattcc gtggcaatac tctgagaatg aaaagcatgg
     1081 catgtttttg atggctttcg agaacaaagc gggacgggga tcaggcatgg tttcaaaggg
     1141 agaagagttg ttcacagggg ttgtaccgat tcttgtagaa cttgacggag atgtcaacgg
     1201 gcacaagttt agtgtaagtg gagaaggcga gggcgatgcg acctatggaa aactgaccct
     1261 taaatttatt tgtactacgg gtaaattgcc cgttccctgg ccgacattgg taactacatt
     1321 cggatatggt ttgatgtgct ttgcaaggta tccggatcac atgaagcagc atgatttctt
     1381 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     1441 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     1501 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     1561 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     1621 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     1681 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca
     1741 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     1801 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagggatcag gcccgcggct
     1861 tccagtggaa ccagcaacgt ttcagcttta tgtgcctgct cttagcgctc tctggagaga
     1921 ttcagggatc cgagaggcgt tttcacgacg gtccgaattc caactgggag aatccgtgaa
     1981 atattttctg gacaatctcg atcgcatcgg tcaattgaat tatttcccca gcaagcaaga
     2041 tatccttctg gctcgcaagg cgacgaaagg tatagtagag catgacttcg tcataaaaaa
     2101 aatccctttt aaaatggttg atgtaggagg ccagaggtca cagcggcaaa aatggtttca
     2161 atgcttcgac ggaatcactt ctatcttgtt catggtgtcc agttctgaat acgaccaagt
     2221 gctcatggag gaccggagga ccaatcgact ggtcgaaagc atgaacatat tcgagactat
     2281 agtgaataac aagttgttct tcaatgtaag tattatcctt tttctcaata aaatggatct
     2341 tttggtagag aaggttaaaa ccgtatcaat taaaaagcat tttcccgatt ttagaggtga
     2401 cccccaccaa ctggaagacg tacagcgata tctggtccag tgtttcgacc ggaaacgccg
     2461 caataggtcc aagcccctgt ttcaccattt tacgactgcg attgataccg aaaacgttag
     2521 gtttgttttc catgctgtga aagatacaat acttcaagag aacctcaagg acattatgct
     2581 ccaatgagcg gccgccgccc ctctccctcc ccccccccta acgttactgg ccgaagccgc
     2641 ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt
     2701 ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt
     2761 tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg
     2821 gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca
     2881 cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg
     2941 gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc
     3001 tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct
     3061 gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta
     3121 ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata agcttgccac
     3181 aacccgtacc aaagatgggg agtagcaaga gcaagacgtc gaattccgga atcaagcttc
     3241 tgattattgg cccagaggaa gattatgacc caggttattt caacaatgag agtgacatca
     3301 tcttccaaga tcttgaaaaa ctgaagtcac atccagctta cttggtagtt tttctacgtt
     3361 acatcctctc tcaggcagac cctggccccc tgctttttta tttgtgttca gaagtttatc
     3421 aacagacaaa tcccaaagat tcccgaagtc tggggaaaga catctggaac attttcctgg
     3481 agaaaaatgc gcctctcaga gtgaagatcc ctgagatgtt gcaggctgaa attgacctac
     3541 gcctgcggaa caatgaggac cctcgcaatg tgctctgtga agctcaggag gcagtcatgc
     3601 tggaaatcca ggagcagatc aacgactaca gatccaagcg tactctgggc ctgggcagcc
     3661 tctatggtga aaatgacctg ctaggcctgg atggggaccc tcttcgagaa cgccaaatgg
     3721 ctgagaagca gctggctgcc cttggagata tcttgtccaa atatgaggaa gatcggagtg
     3781 cccccatgga ctttgctgtt aataccttta tgagccacgc tgggatccgt cttcgggagt
     3841 ctcgaggcgg atcaggcgga tccatggtct tcacactcga agatttcgtt ggggactggc
     3901 gacagacagc cggctacaac ctggaccaag tccttgaaca gggaggtgtg tccagtttgt
     3961 ttcagaatct cggggtgtcc gtaactccga tccaaaggat tgtcctgagc ggtgaaaatg
     4021 ggctgaagat cgacatccat gtcatcatcc cgtatgaagg tctgagcggc gaccaaatgg
     4081 gccagatcga aaaaattttt aaggtggtgt accctgtgga tgatcatcac tttaaggtga
     4141 tcctgcacta tggcacactg gtaatcgacg gggttacgcc gaacatgatc gactatttcg
     4201 gacggccgta tgaaggcatc gccgtgttcg acggcaaaaa gatcactgta acagggaccc
     4261 tgtggaacgg caacaaaatt atcgacgagc gcctgatcaa ccccgacggc tccctgctgt
     4321 tccgagtaac catcaacgga gtgaccggct ggcggctgtg cgaacgcatt ctggcgtaaa
     4381 cgcgtctgga acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     4441 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     4501 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     4561 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     4621 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     4681 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     4741 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     4801 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     4861 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     4921 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     4981 c
//
