LOCUS       sequence-396725-        4460 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 4460)
  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 4460)
  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-hSyn-Galpha-i1 ONE-GO
FEATURES             Location/Qualifiers
     source          1..4460
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_feature    188..303
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        345..792
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             join(1188..1190,1191..1193,1194..1904)
                     /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(1233..1254)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1494..1513)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1841..1862
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(2620..3194,3195..3197,3198..3206)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2787..2804)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3014..3033
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3217..3237
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSK"
     CDS             3343..3858
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     misc_feature    3872..4460
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3925..3945)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4343..4354)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
       61 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
      121 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg
      181 tatcgccttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
      241 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa
      301 ttttcgggtt tattacaggg acagcagaga tccagtttat cgatagtgca agtgggtttt
      361 aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga cccactggac
      421 aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg gaggggaaac
      481 aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt gccttcgccc
      541 ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac gtcactcgcc
      601 ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc cgccggccca
      661 gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca tctgcgctgc
      721 ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt cgtgtcgtgc
      781 ctgagagcgc agctcgagac tagttctaga gcggccgcgg atccatgggc tgcacactga
      841 gcgctgagga caaggcggcc gtggagcgca gcaagatgat cgaccgcaac ctccgggagg
      901 acggagagaa ggcagcgcgc gaggtcaagc tgctgctgct gggtgctggt gaatccggga
      961 agagcacaat tgtgaagcag atgaaaatta tccacgaggc tggctactca gaggaagagt
     1021 gtaagcagta caaagcagtg gtctacagca acaccatcca gtccatcatt gccatcatta
     1081 gagccatggg gagattgaaa atcgactttg gagacgctgc tcgtgcggat gatgctcgcc
     1141 aactcttcgt gcttgctggg gctgcagagg aaggctttat gaccgcgatg gtgagcaagg
     1201 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     1261 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     1321 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct
     1381 tcggctacgg cctgatgtgc ttcgcccgct accccgacca catgaagcag cacgacttct
     1441 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     1501 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     1561 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     1621 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     1681 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     1741 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc
     1801 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     1861 tgaccgccgc cgggatcact ctcggcatgg acgagctata caaggcggag ctcgccggcg
     1921 tcataaagag actgtggaag gacagcggtg tgcaagcctg cttcaacaga tcccgggagt
     1981 accagctgaa cgattcggcg gcgtactacc tgaatgactt ggacagaata gcacaaccaa
     2041 attacatccc aacccagcag gatgttctca gaactagagt gaaaacgacg ggaattgtgg
     2101 aaacccactt tactttcaaa gatcttcatt ttaaaatgtt tgacgtggga ggccagagat
     2161 cagagcggaa gaagtggatt cactgctttg aaggcgtgac tgccatcatc ttctgtgtgg
     2221 ccctgagtga ctatgacctg gttcttgctg aggatgaaga aatgaaccgg atgcacgaaa
     2281 gcatgaagct gttcgatagc atatgtaaca acaagtggtt tacggacaca tccatcatcc
     2341 ttttcctgaa caagaaggac ctcttcgaag agaagatcaa aaagagtccc ctcacgatat
     2401 gctatccaga atatgcaggc tcaaacacat atgaagaggc ggctgcgtat atccagtgtc
     2461 agtttgaaga cctcaataaa aggaaggaca caaaggaaat ttacacccac ttcacttgcg
     2521 ccacggatac gaagaatgtg cagtttgtgt tcgatgctgt aacggacgtc atcataaaga
     2581 ataacctaaa agactgtggt ctcttctaag cggccgccgc ccctctccct cccccccccc
     2641 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt
     2701 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt
     2761 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt
     2821 cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct
     2881 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt
     2941 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt
     3001 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa
     3061 ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta
     3121 gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa
     3181 aacacgatga taagcttgcc acaacccgta ccaaagatgg ggagtagcaa gagcaagacg
     3241 tcgaattccg gaatcaagct tggtggatct tctcgaggtt actaccatgg tatttgggtg
     3301 ggtgaagaag gtcgactttc tcgaggtggc accggtggat ccatggtctt cacactcgaa
     3361 gatttcgttg gggactggcg acagacagcc ggctacaacc tggaccaagt ccttgaacag
     3421 ggaggtgtgt ccagtttgtt tcagaatctc ggggtgtccg taactccgat ccaaaggatt
     3481 gtcctgagcg gtgaaaatgg gctgaagatc gacatccatg tcatcatccc gtatgaaggt
     3541 ctgagcggcg accaaatggg ccagatcgaa aaaattttta aggtggtgta ccctgtggat
     3601 gatcatcact ttaaggtgat cctgcactat ggcacactgg taatcgacgg ggttacgccg
     3661 aacatgatcg actatttcgg acggccgtat gaaggcatcg ccgtgttcga cggcaaaaag
     3721 atcactgtaa cagggaccct gtggaacggc aacaaaatta tcgacgagcg cctgatcaac
     3781 cccgacggct ccctgctgtt ccgagtaacc atcaacggag tgaccggctg gcggctgtgc
     3841 gaacgcattc tggcgtaaac gcgtctggaa caatcaacct ctggattaca aaatttgtga
     3901 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     3961 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     4021 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     4081 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     4141 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     4201 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     4261 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     4321 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     4381 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     4441 cctttgggcc gcctccccgc
//
