LOCUS       sequence-396726-        4463 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 4463)
  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 4463)
  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-i3 ONE-GO
FEATURES             Location/Qualifiers
     source          1..4463
                     /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(1167..1169,1170..1172,1173..1883)
                     /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(1212..1233)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1473..1492)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1820..1841
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(2623..3197,3198..3200,3201..3209)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2790..2807)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3017..3036
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3220..3240
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSK"
     CDS             3346..3861
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     misc_feature    3875..4463
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3928..3948)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4346..4357)
                     /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 tgcacgttga
      841 gcgccgagga caaggcggcg gtggagcgga gtaagatgat cgaccgcaac ttgcgggagg
      901 acggagagaa agcggccaaa gaagtgaagc tgctgctgct cggcgctgga gaatctggta
      961 aaagtactat tgtgaaacag atgaaaatca ttcatgagga tggctattcc gaggacgagt
     1021 gtaagcagta taaagttgtc gtctacagca ataccattca gtccatcatt gcaatcataa
     1081 gagccatggg acggttgaag attgattttg gggaagctgc cagagcggat gatgcccgac
     1141 agttatttgt tttagctggc agcgctatgg tgagcaaggg cgaggagctg ttcaccgggg
     1201 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     1261 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     1321 gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt cggctacggc ctgatgtgct
     1381 tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     1441 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     1501 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     1561 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     1621 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     1681 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     1741 gccccgtgct gctgcccgac aaccactacc tgagctacca gtccgccctg agcaaagacc
     1801 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     1861 tcggcatgga cgagctgtac aagagcgctg aggaaggagt catgacttca gaactagcag
     1921 gcgtgattaa acgtttatgg cgagatggcg gggtgcaggc atgcttcagc aggtccaggg
     1981 aatatcagct caatgattct gcttcatatt acctaaatga tttggataga atatcccaga
     2041 ccaactacat tccaactcag caagatgttc ttcggacgag agtgaagact acaggcattg
     2101 tggagaccca cttcaccttc aaggagctct acttcaaaat gtttgatgta ggtggccaaa
     2161 gatccgaacg gaaaaagtgg atccactgtt ttgagggagt gacagcaatt atcttttgtg
     2221 tggctctcag tgattacgac cttgttctgg ctgaggacga ggaaatgaac cgaatgcatg
     2281 aaagcatgaa attgtttgac agcatttgta acaacaaatg gtttacagac acttcaatca
     2341 ttctcttcct taataagaaa gacctttttg aggaaaaaat aaagaggagt ccattaacaa
     2401 tctgttatcc agaatacaca ggttccaata cgtacgaaga ggcagctgct tacatccagt
     2461 gccagtttga agatctgaac cgaagaaagg acaccaagga ggtctacact cactttacct
     2521 gtgccactga caccaaaaac gtgcagtttg tttttgatgc tgttacagat gtcatcatta
     2581 aaaacaactt aaaggaatgt gggctttact gagcggccgc cgcccctctc cctccccccc
     2641 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
     2701 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
     2761 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
     2821 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac
     2881 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
     2941 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
     3001 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca
     3061 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt
     3121 ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg
     3181 aaaaacacga tgataagctt gccacaaccc gtaccaaaga tggggagtag caagagcaag
     3241 acgtcgaatt ccggaatcaa gcttggtgga tcttctcgag gttactacca tggtatttgg
     3301 gtgggtgaag aaggtcgact ttctcgaggt ggcaccggtg gatccatggt cttcacactc
     3361 gaagatttcg ttggggactg gcgacagaca gccggctaca acctggacca agtccttgaa
     3421 cagggaggtg tgtccagttt gtttcagaat ctcggggtgt ccgtaactcc gatccaaagg
     3481 attgtcctga gcggtgaaaa tgggctgaag atcgacatcc atgtcatcat cccgtatgaa
     3541 ggtctgagcg gcgaccaaat gggccagatc gaaaaaattt ttaaggtggt gtaccctgtg
     3601 gatgatcatc actttaaggt gatcctgcac tatggcacac tggtaatcga cggggttacg
     3661 ccgaacatga tcgactattt cggacggccg tatgaaggca tcgccgtgtt cgacggcaaa
     3721 aagatcactg taacagggac cctgtggaac ggcaacaaaa ttatcgacga gcgcctgatc
     3781 aaccccgacg gctccctgct gttccgagta accatcaacg gagtgaccgg ctggcggctg
     3841 tgcgaacgca ttctggcgta aacgcgtctg gaacaatcaa cctctggatt acaaaatttg
     3901 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     3961 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     4021 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     4081 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     4141 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     4201 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     4261 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     4321 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     4381 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     4441 ctccctttgg gccgcctccc cgc
//
