LOCUS       sequence-180881-        7371 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A FRET sensor for Gq activation, encoded on a single plasmid, for 
            FLIM.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7371)
  AUTHORS   Mastop M, Bindels DS, Shaner NC, Postma M, Gadella TWJ Jr, Goedhart 
            J
  TITLE     Characterization of a spectrally diverse set of fluorescent proteins
            as FRET acceptors for mTurquoise2.
  JOURNAL   Sci Rep. 2017 Sep 20;7(1):11999. doi: 10.1038/s41598-017-12212-x.
  PUBMED    28931898
REFERENCE   2  (bases 1 to 7371)
  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 Rep."; 
            date: "2017-09-20"; volume: "7(1):11999. doi"; pages: " 
            10.1038/s41598-017-12212-x"
COMMENT     Sequence Label: Gq-sensor[mTq2-sREACh]
FEATURES             Location/Qualifiers
     source          1..7371
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1326)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNWNSVNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     ERVTAAGITLGMDELY"
     primer_bind     complement(1611..1628)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    join(1640..2214,2215..2217,2218..2226)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1807..1824)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2034..2053
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2626..2628,2629..2631,2632..3144)
                     /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
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIE"
     polyA_signal    4159..4280
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4196..4215)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4250..4269
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4287..4742)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4424..4445)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4636..4655
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4769..4873
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4875..5232
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4875..5066
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4898..4918)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5083..5218
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5267..6061
                     /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             5267..6061
                     /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(5321..5340)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5931..5950
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6249..6268)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6293..6340
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6669..7257
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7158..7177
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ctcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccttcggc tacggcctga tgtgcttcgc ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactgg aacagcgtca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gagagtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacatgg ccagcaacaa caccgccagc atagcacaag ccaggaagct ggtagagcag
     1381 cttaagatgg aagccaatat cgacaggata aaggtgtcca aggcagctgc agatttgatg
     1441 gcctactgtg aagcacatgc caaggaagac cccctcctga cccctgttcc ggcttcagaa
     1501 aacccgttta gggagaagaa gtttttctgt gccatccttt aaccgcggcg tacgaggcct
     1561 gcatgctccg gacctgcagg ttcgaagtcg acagatctca attggggccc ctatagtgtc
     1621 acctaaataa ttccgccccc ccctctccct cccccccccc taacgttact ggccgaagcc
     1681 gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt
     1741 ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc
     1801 tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc
     1861 tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc
     1921 cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg
     1981 cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct
     2041 cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat
     2101 ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc
     2161 taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga taatatggcc
     2221 acaaccatga ctctggagtc catcatggcg tgctgcctga gcgaggaggc caaggaagcc
     2281 cggcggatca acgacgagat cgagcggcag ctccgcaggg acaagcggga cgcccgccgg
     2341 gagctcaagc tgctgctgct cgggacagga gagagtggca agagtacgtt tatcaagcag
     2401 atgagaatca tccatgggtc aggatactct gatgaagata aaaggggctt caccaagctg
     2461 gtgtatcaga acatcttcac ggccatgcag gccatgatca gagccatgga cacactcaag
     2521 atcccataca agtatgagca caataaggct catgcacaat tagttcgaga agttgatgtg
     2581 gagaaggtgt ctgcttttga ggatccaagc ggaggcggag gcagcatggt gagcaagggc
     2641 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     2701 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     2761 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     2821 tcctggggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     2881 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     2941 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     3001 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     3061 tactttagcg acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac
     3121 ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag
     3181 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     3241 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     3301 accgccgccg ggatcactct cggcatatac gtagatgcaa taaagagttt atggaatgat
     3361 cctggaatcc aggaatgcta tgatagacga cgagaatatc aattatctga ctctaccaaa
     3421 tactatctta atgacttgga ccgcgtagct gaccctgcct acctgcctac gcaacaagat
     3481 gtgcttagag ttcgagtccc caccacaggg atcatcgaat acccctttga cttacaaagt
     3541 gtcattttca gaatggtcga tgtagggggc caaaggtcag agagaagaaa atggatacac
     3601 tgctttgaaa atgtcacctc tatcatgttt ctagtagcgc ttagtgaata tgatcaagtt
     3661 ctcgtggagt cagacaatga gaaccgaatg gaggaaagca aggctctctt tagaacaatt
     3721 atcacatacc cctggttcca gaactcctcg gttattctgt tcttaaacaa gaaagatctt
     3781 ctagaggaga aaatcatgta ttcccatcta gtcgactact tcccagaata tgatggaccc
     3841 cagagagatg cccaggcagc ccgagaattc attctgaaga tgttcgtgga cctgaaccca
     3901 gacagtgaca aaattatcta ctcccacttc acgtgcgcca cagacaccga gaatatccgc
     3961 tttgtctttg ctgccgtcaa ggacaccatc ctccagttga acctgaagga gtacaatctg
     4021 gtctaactcg agtctagagg gccgcgactc tagatcataa tcagccatac cacatttgta
     4081 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     4141 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4201 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     4261 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     4321 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     4381 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     4441 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     4501 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     4561 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     4621 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     4681 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     4741 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4801 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4861 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     4921 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4981 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5041 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     5101 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     5161 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     5221 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     5281 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     5341 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     5401 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     5461 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     5521 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     5581 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     5641 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     5701 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     5761 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     5821 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     5881 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     5941 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     6001 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     6061 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     6121 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     6181 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     6241 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     6301 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     6361 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     6421 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     6481 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     6541 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6601 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6661 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6721 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6781 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     6841 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6901 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6961 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     7021 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     7081 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     7141 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7201 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7261 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     7321 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
