LOCUS       sequence-396734-        6403 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 6403)
  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 6403)
  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-s-IRESWT-Gbetagamma ONE-GO
FEATURES             Location/Qualifiers
     source          1..6403
                     /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)"
     misc_feature    1978..2551
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2001..2553
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2145..2162)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2372..2391
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2553..2573
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSK"
     CDS             3180..3692
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             3702..3755
                     /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"
     CDS             4776..4829
                     /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"
     CDS             4836..5570
                     /codon_start=1
                     /product="circularly permuted form of the Venus yellow 
                     fluorescent protein (Nagai et al., 2004)"
                     /label=cp173Venus
                     /note="efficient FRET acceptor for CFP"
                     /translation="MDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRD
                     HMVLLEFVTAAGITLGMDELYKGGSGGMVSKGEELFTGVVPILVELDGDVNGHKFSVSG
                     EGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPE
                     GYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHN
                     VYITADKQKNGIKANFKIRHNIE"
     primer_bind     4973..4994
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5097..5118)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5358..5377)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    5815..6403
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5868..5888)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6286..6297)
                     /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 tgcctcggga
      841 acagtaagac cgaggaccag cgcaacgagg agaaggcgca gcgtgaggcc aacaaaaaga
      901 tcgagaagca gctgcagaag gacaagcagg tctaccgggc cacgcaccgc ctgctgctgc
      961 tgggtgctgg agaatctggt aaaagcacca ttgtgaagca gatgaggatc ctgcatgtta
     1021 atgggtttaa tggagacagt gagaaggcaa ccaaagtgca ggacatcaaa aacaacctga
     1081 aagaggcgat tgaaaccatt gtggccgcca tgagcaacct ggtgcccccc gtggagctgg
     1141 ccaaccccga gaaccagttc agagtggact acattctgag tgtgatgaac gtgcctgact
     1201 ttgacttccc tcccgaattc tatgagcatg ccaaggctct gtgggaggat gaaggagtgc
     1261 gtgcctgcta cgaacgctcc aacgagtacc agctgattga ctgtgcccag tacttcctgg
     1321 acaagatcga cgtgatcaag caggctgact atgtgccgag cgatcaggac ctgcttcgct
     1381 gccgtgtcct gacttctgga atctttgaga ccaagttcca ggtggacaaa gtcaacttcc
     1441 acatgtttga cgtgggtggc cagcgcgatg aacgccgcaa gtggatccag tgcttcaacg
     1501 atgtgactgc catcatcttc gtggtggcca gcagcagcta caacatggtc atccgggagg
     1561 acaaccagac caaccgcctg caggaggctc tgaacctctt caagagcatc tggaacaaca
     1621 gatggctgcg caccatctct gtgatcctgt tcctcaacaa gcaagatctg ctcgctgaga
     1681 aagtccttgc tgggaaatcg aagattgagg actactttcc agaatttgct cgctacacta
     1741 ctcctgagga tgctactccc gagcccggag aggacccacg cgtgacccgg gccaagtact
     1801 tcattcgaga tgagtttctg aggatcagca ctgccagtgg agatgggcgt cactactgct
     1861 accctcattt cacctgcgct gtggacactg agaacatccg ccgtgtgttc aacgactgcc
     1921 gtgacatcat tcagcgcatg caccttcgtc agtacgagct gctataagcg gccgccgccc
     1981 ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc
     2041 gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa
     2101 acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat
     2161 gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac
     2221 aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg
     2281 cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt
     2341 tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg
     2401 gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac
     2461 atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga
     2521 cgtggttttc ctttgaaaaa cacgatgata atatggggag tagcaagagc aagacgtcga
     2581 attccggaat caagcttctt gattgcgacc aagaactcta caagaacttc cctctggtga
     2641 tctctgagcg ctggcagcag gaagtggcgg aaacagttta tgaagcagta aatgcagaca
     2701 cggataaaat cgaggccagg aagagagcta aaaataagca gcttggccac gaagaagatt
     2761 acgccctggg aagagactgc atcgtgcacg ggtacatgct gaagctgggg aaccctttcc
     2821 tgacccagtg gcagcgccgc tatttttacc tctttccgaa cagacttgag tggagaggag
     2881 aaggcgagtc gcgacaaagt ttactgacaa tggaacagat tgtgtccgtg gaagaaactc
     2941 agattaaaga caaaaagtgc attttgttga gaataaaagg agggaagcag ttcgttttgc
     3001 agtgtgagag tgacccagag tttgtgcagt ggaagaaaga gctgacggag acattcatgg
     3061 aggcccagcg gctgctacgg cgagccccca agttcctcaa caaatcccgc tcagccgtcg
     3121 tggaactctc aaagcctccc ctctgccata ggaacagcaa cggcctcgga ggtggatcca
     3181 tggtcttcac actcgaagat ttcgttgggg actggcgaca gacagccggc tacaacctgg
     3241 accaagtcct tgaacaggga ggtgtgtcca gtttgtttca gaatctcggg gtgtccgtaa
     3301 ctccgatcca aaggattgtc ctgagcggtg aaaatgggct gaagatcgac atccatgtca
     3361 tcatcccgta tgaaggtctg agcggcgacc aaatgggcca gatcgaaaaa atttttaagg
     3421 tggtgtaccc tgtggatgat catcacttta aggtgatcct gcactatggc acactggtaa
     3481 tcgacggggt tacgccgaac atgatcgact atttcggacg gccgtatgaa ggcatcgccg
     3541 tgttcgacgg caaaaagatc actgtaacag ggaccctgtg gaacggcaac aaaattatcg
     3601 acgagcgcct gatcaacccc gacggctccc tgctgttccg agtaaccatc aacggagtga
     3661 ccggctggcg gctgtgcgaa cgcattctgg cgggatcagg agaaggtaga ggaagtctac
     3721 taacatgcgg ggacgtggag gaaaatcccg gcccaatggg ggagcttgac cagttacggc
     3781 aggaggccga gcaacttaag aaccagattc gagacgccag gaaagcatgt gcagatgcaa
     3841 ctctctctca gatcacaaac aacatcgacc cagtgggaag aatccaaatg cgcacgagga
     3901 ggacactgcg ggggcacctg gccaagatct acgccatgca ctggggcaca gactccaggc
     3961 ttctcgtcag tgcctcgcag gatggtaaac ttatcatctg ggacagctac accaccaaca
     4021 aggtccacgc catccctctg cgctcctcct gggtcatgac ctgtgcatat gccccttctg
     4081 ggaactatgt ggcctgcggt ggcctggata acatttgctc catttacaat ctgaaaactc
     4141 gtgaggggaa cgtgcgcgtg agtcgtgagc tggcaggaca cacaggttac ctgtcctgct
     4201 gccgattcct ggatgacaat cagatcgtca ccagctctgg agacaccacg tgtgccctgt
     4261 gggacatcga gaccggccag cagacgacca cgtttaccgg acacactgga gatgtcatga
     4321 gcctttctct tgctcctgac accagactgt tcgtctctgg tgcttgtgat gcttcagcca
     4381 aactctggga tgtgcgagaa ggcatgtgcc ggcagacctt cactggccac gagtctgaca
     4441 tcaatgccat ttgcttcttt ccaaatggca atgcatttgc cactggctca gacgacgcca
     4501 cctgcaggct gtttgacctt cgtgctgacc aggagctcat gacttactcc catgacaaca
     4561 tcatctgcgg gatcacctct gtctccttct ccaagagcgg gcgcctcctc cttgctgggt
     4621 acgacgactt caactgcaac gtctgggatg cactcaaagc cgaccgggca ggtgtcttgg
     4681 ctgggcatga caaccgcgtc agctgcctgg gcgtgactga cgatggcatg gctgtggcga
     4741 cagggtcctg ggatagcttc ctcaagatct ggaacgaggg cagaggaagt cttctaacat
     4801 gcggtgacgt ggaggagaat cccggcccta ccggtatgga cggcggcgtg cagctcgccg
     4861 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     4921 acctgagcta ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     4981 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagggtg
     5041 gcagcggtgg catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     5101 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     5161 atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc
     5221 cctggcccac cctcgtgacc accctgggct acggcctgca gtgcttcgcc cgctaccccg
     5281 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     5341 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     5401 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     5461 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca
     5521 agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag tccggactca
     5581 gatctatggc cagcaacaac accgccagca tagcacaagc caggaagctg gtagagcagc
     5641 ttaagatgga agccaatatc gacaggataa aggtgtccaa ggcagctgca gatttgatgg
     5701 cctactgtga agcacatgcc aaggaagacc ccctcctgac ccctgttccg gcttcagaaa
     5761 acccgtttag ggagaagaag tttttctgtg ccatccttta aacgcgtctg gaacaatcaa
     5821 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     5881 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     5941 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     6001 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     6061 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     6121 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     6181 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt
     6241 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     6301 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     6361 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgc
//
