LOCUS       sequence-396728-        4835 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 4835)
  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 4835)
  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-q ONE-GO
FEATURES             Location/Qualifiers
     source          1..4835
                     /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(1215..1217,1218..1220,1221..1931)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1260..1281)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1521..1540)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1868..1889
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(2668..3242,3243..3245,3246..3254)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2835..2852)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3062..3081
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3265..3285
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSK"
     CDS             3718..4233
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     misc_feature    4247..4835
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4300..4320)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4718..4729)
                     /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 atccatgact ctggagtcca
      841 tcatggcgtg ctgcctgagc gaggaggcca aggaagcccg gaggatcaac gacgagatcg
      901 agcggcagct gcgcagggac aagcgcgacg cccgccggga actcaagctg ctgctgctgg
      961 ggacagggga gagtggcaag agcaccttca tcaagcagat gaggatcatc cacgggtcgg
     1021 gctactctga cgaagacaag cgcggcttca ccaagctggt gtatcagaac atcttcacgg
     1081 ccatgcaggc catgatcaga gcgatggaca cactcaagat cccatacaag tatgaacaca
     1141 ataaggctca tgcacaattg gttcgagagg ttgatgtgga gaaggtgtct gctttttcag
     1201 gaggtggcgg atccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     1261 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     1321 gcgatgccac ctacggcaag ctgaccctga agctgatctg caccaccggc aagctgcccg
     1381 tgccctggcc caccctcgtg accaccctgg gctacggcct gcagtgcttc gcccgctacc
     1441 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     1501 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     1561 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     1621 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcaccgccg
     1681 acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggcg
     1741 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     1801 tgcccgacaa ccactacctg agctaccagt ccaaactgag caaagacccc aacgagaagc
     1861 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     1921 agctgtacaa gtctggcggt ggaggatccg agaatccata tgtagatgca ataaagagct
     1981 tgtggaatga tcctggaatc caggagtgct acgacagacg acgggaatat cagttatctg
     2041 actctaccaa atactatctg aatgacttgg accgtgtagc cgaccctgag tacatgccga
     2101 cagagcaaga cgtgcttaga gttcgagtcc ccactacagg gatcatcgaa tacccctttg
     2161 acttacaaag tgtcattttc agaatggtcg atgtaggggg ccaaaggtca gagagaagaa
     2221 aatggataca ctgctttgaa aatgtcacct ccatcatgtt tctagtagcg cttagcgaat
     2281 atgatcaagt tcttgtggag tcagacaatg agaaccgcat ggaggagagc aaagcactct
     2341 ttagaacaat tatcacctac ccctggttcc agaactcctc tgtgattctg ttcttaaaca
     2401 agaaagatct tctagaggag aaaatcatgt attcccacct agtcgactac ttcccagaat
     2461 atgatggacc ccagagagat gcccaggcag ctcgagaatt catcctgaaa atgttcgtgg
     2521 acctgaaccc cgacagtgac aaaatcatct actcccactt cacgtgcgcc acagataccg
     2581 agaacatccg cttcgtcttt gcagccgtca aggacaccat cctgcagctg aacctgaagg
     2641 agtacaatct ggtctaagcg gccgccgccc ctctccctcc ccccccccta acgttactgg
     2701 ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt
     2761 gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc
     2821 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc
     2881 agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg
     2941 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc
     3001 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa
     3061 atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg
     3121 tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa
     3181 aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata
     3241 agcttgccac aacccgtacc aaagatgggg agtagcaaga gcaagacgtc gaattccgga
     3301 atcaagctta agtacctgga ggaccggggc gaggtgactt ttgagaagat cttctcccag
     3361 aagctggggt acctgctttt ccgagacttc tgcctgaagc acctggagga ggccaagccc
     3421 ttggtagagt tctacgagga gatcaagaaa tacgagaagc tggagacaga ggaggagcgc
     3481 ctggtctgca gccgagagat cttcgacacg tacatcatga aggagctgct ggcctgctca
     3541 catcctttct cgaagagcgc cattgagcac gtccagggcc atctggtgaa gaagcaggtg
     3601 cctccggatc tcttccagcc atatattgaa gaaatttgcc agaacctccg aggagacgtg
     3661 ttccagaaat tcatcgagag cgataaattc acacggtttt gccagtggaa gggatccatg
     3721 gtcttcacac tcgaagattt cgttggggac tggcgacaga cagccggcta caacctggac
     3781 caagtccttg aacagggagg tgtgtccagt ttgtttcaga atctcggggt gtccgtaact
     3841 ccgatccaaa ggattgtcct gagcggtgaa aatgggctga agatcgacat ccatgtcatc
     3901 atcccgtatg aaggtctgag cggcgaccaa atgggccaga tcgaaaaaat ttttaaggtg
     3961 gtgtaccctg tggatgatca tcactttaag gtgatcctgc actatggcac actggtaatc
     4021 gacggggtta cgccgaacat gatcgactat ttcggacggc cgtatgaagg catcgccgtg
     4081 ttcgacggca aaaagatcac tgtaacaggg accctgtgga acggcaacaa aattatcgac
     4141 gagcgcctga tcaaccccga cggctccctg ctgttccgag taaccatcaa cggagtgacc
     4201 ggctggcggc tgtgcgaacg cattctggcg taaacgcgtc tggaacaatc aacctctgga
     4261 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     4321 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     4381 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     4441 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     4501 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     4561 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     4621 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     4681 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     4741 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     4801 gacgagtcgg atctcccttt gggccgcctc cccgc
//
