LOCUS       sequence-332869-        8238 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes a BRET-based activity sensor for the heterotrimeric G 
            protein Gq. Composed of the subunits G alpha q (GNAQ) tagged with 
            NanoLuciferase, G beta 3 (GNB3) and Venus-tagged G gamma 9 (GNG9). .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8238)
  AUTHORS   Schihada H, Shekhani R, Schulte G
  TITLE     Quantitative assessment of constitutive G protein-coupled receptor 
            activity with BRET-based G protein biosensors.
  JOURNAL   Sci Signal. 2021 Sep 7;14(699):eabf1653. doi: 
            10.1126/scisignal.abf1653. Epub 2021 Sep 7.
  PUBMED    34516756
REFERENCE   2  (bases 1 to 8238)
  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: "Sci 
            Signal."; date: "2021-09-7"; volume: "14(699):eabf1653. doi"; pages:
            " 10.1126/scisignal.abf1653. Epub 2021 Sep 7"
COMMENT     Sequence Label: Gq-CASE
FEATURES             Location/Qualifiers
     source          1..8238
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1557..1610
                     /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             1617..2351
                     /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     1754..1775
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1878..1899)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2139..2158)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(2598..2615)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    2627..3200
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2794..2811)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3021..3040
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3604..4113
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     polyA_signal    4966..5087
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5003..5022)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5057..5076
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5094..5549)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5231..5252)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5443..5462
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5576..5680
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5682..6039
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5682..5873
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5705..5725)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5890..6025
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6074..6868
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6074..6868
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6128..6147)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6738..6757
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7056..7075)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7100..7147
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7476..8064
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7965..7984
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcatgg
      541 gggagatgga gcaactgcgt caggaagcgg agcagctcaa gaagcagatt gcagatgcca
      601 ggaaagcctg tgctgacgtt actctggcag agctggtgtc tggcctagag gtggtgggac
      661 gagtccagat gcggacgcgg cggacgttaa ggggacacct ggccaagatt tacgccatgc
      721 actgggccac tgattctaag ctgctggtaa gtgcctcgca agatgggaag ctgatcgtgt
      781 gggacagcta caccaccaac aaggtgcacg ccatcccact gcgctcctcc tgggtcatga
      841 cctgtgccta tgccccatca gggaactttg tggcgtgtgg ggggctggac aacatgtgtt
      901 ccatctacaa cctcaaatcc cgtgagggca atgtcaaggt cagccgggag ctttctgctc
      961 acacaggtta tctctcctgc tgccgcttcc tggatgacaa caatattgtg accagctcgg
     1021 gggacaccac gtgtgccttg tgggacattg agactgggca gcagaagact gtatttgtgg
     1081 gacacacggg tgactgcatg agcctggctg tgtctcctga cttcaatctc ttcatttcgg
     1141 gggcctgtga tgccagtgcc aagctctggg atgtgcgaga ggggacctgc cgacagactt
     1201 tcactggcca cgagtcggac atcaacgcca tctgtttctt ccccaatgga gaggccatct
     1261 gcacgggctc ggatgacgct tcctgccgct tgtttgacct gcgggcagac caggagctga
     1321 tctgcttctc ccacgagagc atcatctgcg gcatcacgtc tgtggccttc tccctcagtg
     1381 gccgcctact attcgctggc tacgacgact tcaactgcaa tgtctgggac tccatgaagt
     1441 ctgagcgtgt gggcatcctc tctggccacg ataacagggt gagctgcctg ggagtcacag
     1501 ctgacgggat ggctgtggcc acaggttcct gggacagctt cctcaaaatc tggaacgagg
     1561 gcagaggaag tcttctaaca tgcggtgacg tggaggagaa tcccggccct accggtatgg
     1621 acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     1681 tgctgctgcc cgacaaccac tacctgagct accagtccgc cctgagcaaa gaccccaacg
     1741 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     1801 tggacgagct gtacaagggt ggcagcggtg gcatggtgag caagggcgag gagctgttca
     1861 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     1921 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ctgatctgca
     1981 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgggc tacggcctgc
     2041 agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     2101 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     2161 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     2221 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     2281 acgtctatat caccgccgac aaacagaaga acggcatcaa ggccaacttc aagatccgcc
     2341 acaacatcga gtccggactc agatctgccc aggatctcag cgagaaggac ctgttgaaga
     2401 tggaggtgga gcagctgaag aaagaagtga aaaacacaag aattccgatt tccaaagcgg
     2461 gaaaggaaat caaggagtac gtggaggccc aagcaggaaa cgatcctttt ctcaaaggca
     2521 tccctgagga caagaatccc ttcaaggaga aaggtggctg tctgataagc taagtcgaca
     2581 gatctcaatt ggggccccta tagtgtcacc taaataattc cgcccccccc tctccctccc
     2641 ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata
     2701 tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg
     2761 tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt
     2821 tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag
     2881 cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc
     2941 cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga
     3001 tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg
     3061 cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat
     3121 gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc
     3181 tttgaaaaac acgatgataa tggatccatg actctggagt ccatcatggc gtgctgcctg
     3241 agcgaggagg ccaaggaagc ccggcggatc aacgacgaga tcgagcggca gctccgcagg
     3301 gacaagcggg acgcccgccg ggagctcaag ctgctgctgc tcgggacagg agagagtggc
     3361 aagagtacgt ttatcaagca gatgagaatc atccatgggt caggatactc tgatgaagat
     3421 aaaaggggct tcaccaagct ggtgtatcag aacatcttca cggccatgca ggccatgatc
     3481 agagccatgg acacactcaa gatcccatac aagtatgagc acaataaggc tcatgcacaa
     3541 ttagttcgag aagttgatgt ggagaaggtg tctgctttta gcggtggagg cgggacgcgt
     3601 tctgtcttca cactcgaaga tttcgttggg gactggcgac agacagccgg ctacaacctg
     3661 gaccaagtcc ttgaacaggg aggtgtgtcc agtttgtttc agaatctcgg ggtgtccgta
     3721 actccgatcc aaaggattgt cctgagcggt gaaaatgggc tgaagatcga catccatgtc
     3781 atcatcccgt atgaaggtct gagcggcgac caaatgggcc agatcgaaaa aatttttaag
     3841 gtggtgtacc ctgtggatga tcatcacttt aaggtgatcc tgcactatgg cacactggta
     3901 atcgacgggg ttacgccgaa catgatcgac tatttcggac ggccgtatga aggcatcgcc
     3961 gtgttcgacg gcaaaaagat cactgtaaca gggaccctgt ggaacggcaa caaaattatc
     4021 gacgagcgcc tgatcaaccc cgacggctcc ctgctgttcc gagtaaccat caacggagtg
     4081 accggctggc ggctgtgcga acgcattctg gccaccggtg gtggcggatc cgagaatcca
     4141 tatgtagatg caataaagag tttatggaat gatcctggaa tccaggaatg ctatgataga
     4201 cgacgagaat atcaattatc tgactctacc aaatactatc ttaatgactt ggaccgcgta
     4261 gctgaccctg cctacctgcc tacgcaacaa gatgtgctta gagttcgagt ccccaccaca
     4321 gggatcatcg aatacccctt tgacttacaa agtgtcattt tcagaatggt cgatgtaggg
     4381 ggccaaaggt cagagagaag aaaatggata cactgctttg aaaatgtcac ctctatcatg
     4441 tttctagtag cgcttagtga atatgatcaa gttctcgtgg agtcagacaa tgagaaccga
     4501 atggaggaaa gcaaggctct ctttagaaca attatcacat acccctggtt ccagaactcc
     4561 tcggttattc tgttcttaaa caagaaagat cttcttgagg agaaaatcat gtattcccat
     4621 ctagtggact acttcccaga atatgatgga ccccagagag atgcccaggc agcccgagaa
     4681 ttcattctga agatgttcgt ggacctgaac ccagacagtg acaaaattat ctactcccac
     4741 ttcacgtgcg ccacagacac cgagaatatc cgctttgtct ttgctgccgt caaggacacc
     4801 atcctccagt tgaacctgaa ggagtacaat ctggtctaat ctagagggcc gcgactctag
     4861 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     4921 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     4981 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5041 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt
     5101 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     5161 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     5221 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     5281 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     5341 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     5401 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     5461 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     5521 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     5581 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5641 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     5701 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     5761 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     5821 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     5881 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     5941 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     6001 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     6061 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     6121 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     6181 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     6241 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     6301 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     6361 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     6421 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     6481 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     6541 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     6601 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     6661 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     6721 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     6781 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     6841 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     6901 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     6961 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     7021 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     7081 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     7141 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     7201 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     7261 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     7321 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     7381 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     7441 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     7501 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     7561 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     7621 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     7681 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     7741 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     7801 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     7861 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     7921 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     7981 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     8041 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     8101 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     8161 cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag
     8221 cccatatatg gagttccg
//
