LOCUS       sequence-394361-        4333 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 4333)
  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 4333)
  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-i1 ONE-GO
FEATURES             Location/Qualifiers
     source          1..4333
                     /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(1061..1063,1064..1066,1067..1777)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1106..1127)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1367..1386)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1714..1735
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(2493..3067,3068..3070,3071..3079)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2660..2677)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2887..2906
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3090..3110
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSK"
     CDS             3216..3731
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     misc_feature    3745..4333
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3798..3818)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4216..4227)
                     /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 ggctgcacac tgagcgctga
      721 ggacaaggcg gccgtggagc gcagcaagat gatcgaccgc aacctccggg aggacggaga
      781 gaaggcagcg cgcgaggtca agctgctgct gctgggtgct ggtgaatccg ggaagagcac
      841 aattgtgaag cagatgaaaa ttatccacga ggctggctac tcagaggaag agtgtaagca
      901 gtacaaagca gtggtctaca gcaacaccat ccagtccatc attgccatca ttagagccat
      961 ggggagattg aaaatcgact ttggagacgc tgctcgtgcg gatgatgctc gccaactctt
     1021 cgtgcttgct ggggctgcag aggaaggctt tatgaccgcg atggtgagca agggcgagga
     1081 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     1141 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     1201 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttcggcta
     1261 cggcctgatg tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     1321 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     1381 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     1441 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     1501 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     1561 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     1621 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccgc
     1681 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     1741 cgccgggatc actctcggca tggacgagct atacaaggcg gagctcgccg gcgtcataaa
     1801 gagactgtgg aaggacagcg gtgtgcaagc ctgcttcaac agatcccggg agtaccagct
     1861 gaacgattcg gcggcgtact acctgaatga cttggacaga atagcacaac caaattacat
     1921 cccaacccag caggatgttc tcagaactag agtgaaaacg acgggaattg tggaaaccca
     1981 ctttactttc aaagatcttc attttaaaat gtttgacgtg ggaggccaga gatcagagcg
     2041 gaagaagtgg attcactgct ttgaaggcgt gactgccatc atcttctgtg tggccctgag
     2101 tgactatgac ctggttcttg ctgaggatga agaaatgaac cggatgcacg aaagcatgaa
     2161 gctgttcgat agcatatgta acaacaagtg gtttacggac acatccatca tccttttcct
     2221 gaacaagaag gacctcttcg aagagaagat caaaaagagt cccctcacga tatgctatcc
     2281 agaatatgca ggctcaaaca catatgaaga ggcggctgcg tatatccagt gtcagtttga
     2341 agacctcaat aaaaggaagg acacaaagga aatttacacc cacttcactt gcgccacgga
     2401 tacgaagaat gtgcagtttg tgttcgatgc tgtaacggac gtcatcataa agaataacct
     2461 aaaagactgt ggtctcttct aagcggccgc cgcccctctc cctccccccc ccctaacgtt
     2521 actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc
     2581 atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc
     2641 attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag
     2701 gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg
     2761 cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat
     2821 acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga
     2881 gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc
     2941 cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg
     3001 ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga
     3061 tgataagctt gccacaaccc gtaccaaaga tggggagtag caagagcaag acgtcgaatt
     3121 ccggaatcaa gcttggtgga tcttctcgag gttactacca tggtatttgg gtgggtgaag
     3181 aaggtcgact ttctcgaggt ggcaccggtg gatccatggt cttcacactc gaagatttcg
     3241 ttggggactg gcgacagaca gccggctaca acctggacca agtccttgaa cagggaggtg
     3301 tgtccagttt gtttcagaat ctcggggtgt ccgtaactcc gatccaaagg attgtcctga
     3361 gcggtgaaaa tgggctgaag atcgacatcc atgtcatcat cccgtatgaa ggtctgagcg
     3421 gcgaccaaat gggccagatc gaaaaaattt ttaaggtggt gtaccctgtg gatgatcatc
     3481 actttaaggt gatcctgcac tatggcacac tggtaatcga cggggttacg ccgaacatga
     3541 tcgactattt cggacggccg tatgaaggca tcgccgtgtt cgacggcaaa aagatcactg
     3601 taacagggac cctgtggaac ggcaacaaaa ttatcgacga gcgcctgatc aaccccgacg
     3661 gctccctgct gttccgagta accatcaacg gagtgaccgg ctggcggctg tgcgaacgca
     3721 ttctggcgta aacgcgtctg gaacaatcaa cctctggatt acaaaatttg tgaaagattg
     3781 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     3841 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     3901 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     3961 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     4021 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     4081 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag
     4141 ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     4201 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     4261 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     4321 gccgcctccc cgc
//
