LOCUS       sequence-396727-        4496 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 4496)
  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 4496)
  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-oA ONE-GO
FEATURES             Location/Qualifiers
     source          1..4496
                     /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(1203..1205,1206..1208,1209..1919)
                     /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(1248..1269)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1509..1528)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1856..1877
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(2656..3230,3231..3233,3234..3242)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2823..2840)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3050..3069
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3253..3273
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSK"
     CDS             3379..3894
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     misc_feature    3908..4496
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3961..3981)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4379..4390)
                     /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 atccatggga tgtactctga
      841 gcgcagagga gagagccgcc ctcgagcgga gcaaggcgat tgagaaaaat ctcaaagaag
      901 atggcatcag cgccgccaaa gacgtgaaat tactcctgct gggggctgga gaatcaggaa
      961 aaagcaccat tgtgaagcag atgaagatca tccatgaaga tggcttctct ggagaagacg
     1021 taaagcagta caagcctgtc gtctacagca acaccatcca gtctctggca gccattgtgc
     1081 gggccatgga tactctgggc gtggagtatg gtgacaagga gaggaaggca gactccaaga
     1141 tggtgtgtga cgtggtgagt cgcatggagg acactgaacc aagatccggt gggggcggga
     1201 gcatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     1261 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     1321 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     1381 ccctcgtgac caccttcggc tacggcctga tgtgcttcgc ccgctacccc gaccacatga
     1441 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     1501 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     1561 tggtgaaccg cattgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     1621 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     1681 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     1741 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     1801 actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     1861 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     1921 ccggaggtgg aggatctttc tctgcagaac tgctttctgc catgatgcga ctctggggcg
     1981 actcggggat ccaggagtgc ttcaaccgat ctcgggagta tcagctcaac gactctgcca
     2041 aatactacct ggacagcttg gatcggattg gagctgcaga atacatgccc accgagcagg
     2101 acatcctccg aaccagggtc aaaacaactg gcatcgtaga aacccacttc accttcaaga
     2161 acctccactt caggctgttt gacgttgggg gccagcgatc tgaacgtaag aagtggatcc
     2221 actgcttcga ggatgtcacg gccatcatct tctgtgtcgc actcagcggc tatgaccagg
     2281 tgctccacga ggacgaaacc acgaaccgca tgcacgagtc tctcatgctc ttcgactcca
     2341 tctgtaacaa caagtttttc atcgatacct ccatcattct cttcctcaac aagaaagacc
     2401 tctttggcga gaagattaag aagtcaccct tgaccatctg ctttcctgaa tacccaggct
     2461 ccaacaccta tgaagacgca gctgcctaca tccaaacaca gtttgaaagc aaaaaccgct
     2521 cacccaacaa agaaatttac tgtcacatga cttgtgccac agacacgaat aatatccagg
     2581 tggtattcga cgccgtcacc gacatcatca ttgccaacaa tctccggggc tgtggcttgt
     2641 actgagcggc cgccgcccct ctccctcccc cccccctaac gttactggcc gaagccgctt
     2701 ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg
     2761 caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc
     2821 ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga
     2881 agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc
     2941 tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc
     3001 acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc
     3061 aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga
     3121 tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg
     3181 ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataag cttgccacaa
     3241 cccgtaccaa agatggggag tagcaagagc aagacgtcga attccggaat caagcttggt
     3301 ggatcttctc gaggttacta ccatggtatt tgggtgggtg aagaaggtcg actttctcga
     3361 ggtggcaccg gtggatccat ggtcttcaca ctcgaagatt tcgttgggga ctggcgacag
     3421 acagccggct acaacctgga ccaagtcctt gaacagggag gtgtgtccag tttgtttcag
     3481 aatctcgggg tgtccgtaac tccgatccaa aggattgtcc tgagcggtga aaatgggctg
     3541 aagatcgaca tccatgtcat catcccgtat gaaggtctga gcggcgacca aatgggccag
     3601 atcgaaaaaa tttttaaggt ggtgtaccct gtggatgatc atcactttaa ggtgatcctg
     3661 cactatggca cactggtaat cgacggggtt acgccgaaca tgatcgacta tttcggacgg
     3721 ccgtatgaag gcatcgccgt gttcgacggc aaaaagatca ctgtaacagg gaccctgtgg
     3781 aacggcaaca aaattatcga cgagcgcctg atcaaccccg acggctccct gctgttccga
     3841 gtaaccatca acggagtgac cggctggcgg ctgtgcgaac gcattctggc gtaaacgcgt
     3901 ctggaacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     3961 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     4021 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     4081 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     4141 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     4201 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     4261 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     4321 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     4381 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     4441 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgc
//
