LOCUS       sequence-273226-        8487 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A FRET sensor for Gq activation, encoded on a single plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8487)
  AUTHORS   Goedhart J, van Weeren L, Adjobo-Hermans MJ, Elzenaar I, Hink MA, 
            Gadella TW Jr
  TITLE     Quantitative co-expression of proteins at the single cell 
            level--application to a multimeric FRET sensor.
  JOURNAL   PLoS One. 2011;6(11):e27321. doi: 10.1371/journal.pone.0027321. Epub
            2011 Nov 17.
  PUBMED    22114669
REFERENCE   2  (bases 1 to 8487)
  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: "PLoS One.";
            date: "2011"; volume: "6(11):e27321. doi"; pages: " 
            10.1371/journal.pone.0027321. Epub 2011 Nov 17"
COMMENT     Sequence Label: Gq sensor
FEATURES             Location/Qualifiers
     source          1..8487
                     /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"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1573..1626
                     /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             1633..2367
                     /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     complement(2667..2684)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    join(2696..3270,3271..3273,3274..3282)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2863..2880)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3090..3109
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3682..3684,3685..3687,3688..4200)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISDNVYITADKQKNGIK
                     ANFKIRHNIE"
     polyA_signal    5215..5336
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5252..5271)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5306..5325
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5343..5798)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5480..5501)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5692..5711
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5825..5929
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5931..6288
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5931..6122
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5954..5974)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6139..6274
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6323..7117
                     /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             6323..7117
                     /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(6377..6396)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6987..7006
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7305..7324)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7349..7396
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7725..8313
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8214..8233
                     /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 gctagcgcta
      541 ccggtcgcca ccatggggga gcttgaccag ttacggcagg aggccgagca acttaagaac
      601 cagattcgag acgccaggaa agcatgtgca gatgcaactc tctctcagat cacaaacaac
      661 atcgacccag tgggaagaat ccaaatgcgc acgaggagga cactgcgggg gcacctggcc
      721 aagatctacg ccatgcactg gggcacagac tccaggcttc tcgtcagtgc ctcgcaggat
      781 ggtaaactta tcatctggga cagctacacc accaacaagg tccacgccat ccctctgcgc
      841 tcctcctggg tcatgacctg tgcatatgcc ccttctggga actatgtggc ctgcggtggc
      901 ctggataaca tttgctccat ttacaatctg aaaactcgtg aggggaacgt gcgcgtgagt
      961 cgtgagctgg caggacacac aggttacctg tcctgctgcc gattcctgga tgacaatcag
     1021 atcgtcacca gctctggaga caccacgtgt gccctgtggg acatcgagac cggccagcag
     1081 acgaccacgt ttaccggaca cactggagat gtcatgagcc tttctcttgc tcctgacacc
     1141 agactgttcg tctctggtgc ttgtgatgct tcagccaaac tctgggatgt gcgagaaggc
     1201 atgtgccggc agaccttcac tggccacgag tctgacatca atgccatttg cttctttcca
     1261 aatggcaatg catttgccac tggctcagac gacgccacct gcaggctgtt tgaccttcgt
     1321 gctgaccagg agctcatgac ttactcccat gacaacatca tctgcgggat cacctctgtc
     1381 tccttctcca agagcgggcg cctcctcctt gctgggtacg acgacttcaa ctgcaacgtc
     1441 tgggatgcac tcaaagccga ccgggcaggt gtcttggctg ggcatgacaa ccgcgtcagc
     1501 tgcctgggcg tgactgacga tggcatggct gtggcgacag ggtcctggga tagcttcctc
     1561 aagatctgga acgagggcag aggaagtctt ctaacatgcg gtgacgtgga ggagaatccc
     1621 ggccctaccg gtatggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc
     1681 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca gtccgccctg
     1741 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     1801 gggatcactc tcggcatgga cgagctgtac aagggtggca gcggtggcat ggtgagcaag
     1861 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     1921 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     1981 ctgaagctga tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     2041 ctgggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
     2101 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     2161 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     2221 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     2281 aactacaaca gccacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc
     2341 aacttcaaga tccgccacaa catcgagtcc ggactcagat ctatggccag caacaacacc
     2401 gccagcatag cacaagccag gaagctggta gagcagctta agatggaagc caatatcgac
     2461 aggataaagg tgtccaaggc agctgcagat ttgatggcct actgtgaagc acatgccaag
     2521 gaagaccccc tcctgacccc tgttccggct tcagaaaacc cgtttaggga gaagaagttt
     2581 ttctgtgcca tcctttaacc gcggcgtacg aggcctgcat gctccggacc tgcaggttcg
     2641 aagtcgacag atctcaattg gggcccctat agtgtcacct aaataattcc gcccccccct
     2701 ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
     2761 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     2821 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     2881 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
     2941 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     3001 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     3061 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
     3121 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat
     3181 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg
     3241 tggttttcct ttgaaaaaca cgatgataat atggccacaa ccatgactct ggagtccatc
     3301 atggcgtgct gcctgagcga ggaggccaag gaagcccggc ggatcaacga cgagatcgag
     3361 cggcagctcc gcagggacaa gcgggacgcc cgccgggagc tcaagctgct gctgctcggg
     3421 acaggagaga gtggcaagag tacgtttatc aagcagatga gaatcatcca tgggtcagga
     3481 tactctgatg aagataaaag gggcttcacc aagctggtgt atcagaacat cttcacggcc
     3541 atgcaggcca tgatcagagc catggacaca ctcaagatcc catacaagta tgagcacaat
     3601 aaggctcatg cacaattagt tcgagaagtt gatgtggaga aggtgtctgc ttttgaggat
     3661 ccaagcggag gcggaggcag catggtgagc aagggcgagg agctgttcac cggggtggtg
     3721 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     3781 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     3841 ctgcccgtgc cctggcccac cctcgtgacc accctgtcct ggggcgtgca gtgcttcgcc
     3901 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     3961 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     4021 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     4081 gacggcaaca tcctggggca caagctggag tacaactaca tcagcgacaa cgtctatatc
     4141 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
     4201 gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     4261 gtgctgctgc ccgacaacca ctacctgagc acccagtcca agctgagcaa agaccccaac
     4321 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     4381 atatatgtag atgcaataaa gagtttatgg aatgatcctg gaatccagga atgctatgat
     4441 agacgacgag aatatcaatt atctgactct accaaatact atcttaatga cttggaccgc
     4501 gtagctgacc ctgcctacct gcctacgcaa caagatgtgc ttagagttcg agtccccacc
     4561 acagggatca tcgaataccc ctttgactta caaagtgtca ttttcagaat ggtcgatgta
     4621 gggggccaaa ggtcagagag aagaaaatgg atacactgct ttgaaaatgt cacctctatc
     4681 atgtttctag tagcgcttag tgaatatgat caagttctcg tggagtcaga caatgagaac
     4741 cgaatggagg aaagcaaggc tctctttaga acaattatca catacccctg gttccagaac
     4801 tcctcggtta ttctgttctt aaacaagaaa gatcttctag aggagaaaat catgtattcc
     4861 catctagtcg actacttccc agaatatgat ggaccccaga gagatgccca ggcagcccga
     4921 gaattcattc tgaagatgtt cgtggacctg aacccagaca gtgacaaaat tatctactcc
     4981 cacttcacgt gcgccacaga caccgagaat atccgctttg tctttgctgc cgtcaaggac
     5041 accatcctcc agttgaacct gaaggagtac aatctggtct aactcgagtc tagagggccg
     5101 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     5161 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     5221 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5281 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
     5341 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     5401 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     5461 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     5521 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     5581 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     5641 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     5701 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     5761 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     5821 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     5881 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     5941 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     6001 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     6061 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     6121 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     6181 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     6241 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     6301 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     6361 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     6421 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     6481 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     6541 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     6601 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     6661 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     6721 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     6781 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     6841 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     6901 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     6961 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     7021 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     7081 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     7141 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     7201 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     7261 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     7321 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     7381 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     7441 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     7501 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     7561 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     7621 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     7681 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     7741 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     7801 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     7861 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     7921 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     7981 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     8041 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     8101 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     8161 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     8221 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     8281 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     8341 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     8401 gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt
     8461 agttcatagc ccatatatgg agttccg
//
