LOCUS       sequence-208325-        6729 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  A JNK biosensor based on fluorescence resonance energy transfer 
            (FRET) and those based on bioluminescence energy transfer (BRET).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6729)
  AUTHORS   Komatsu N, Terai K, Imanishi A, Kamioka Y, Sumiyama K, Jin T, Okada 
            Y, Nagai T, Matsuda M
  TITLE     A platform of BRET-FRET hybrid biosensors for optogenetics, chemical
            screening, and in vivo imaging.
  JOURNAL   Sci Rep. 2018 Jun 12;8(1):8984. doi: 10.1038/s41598-018-27174-x.
  PUBMED    29895862
REFERENCE   2  (bases 1 to 6729)
  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: "2018-06-12"; volume: "8(1):8984. doi"; pages: " 
            10.1038/s41598-018-27174-x"
COMMENT     Sequence Label: pCAGGS-6020nes
FEATURES             Location/Qualifiers
     source          1..6729
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             join(3..5,6..8,9..716)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLLCTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALFKDPNEKRDHMVLL
                     EFLTAAGITEGMNELY"
     CDS             1152..1499
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             3204..3230
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     complement(3246..3265)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3311..3366
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3365..3384)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3727..3743)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3727..3743)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3740..3762)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3751..3767
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3775..3781,3782..3799,3800..3805))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3820..3841
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3900..4095
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3946..4081
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4101..4235
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4138..4157)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4192..4211
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4303..4320)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4474..5062)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4554..4573)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5233..6024,6025..6093))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5856..5875
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6094..6198)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6233..6612
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gcatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
       61 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
      121 acggcaagct gaccctgaag cttctatgca ccaccggcaa gctgcccgtg ccctggccca
      181 ccctcgtgac caccctgggc tacggcctgc agtgcttcgc ccgctacccc gaccacatga
      241 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
      301 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
      361 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
      421 acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac aagcagaaga
      481 acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg
      541 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
      601 actacctgag ctaccagtcc gccctgttca aagaccccaa cgagaagcgc gatcacatgg
      661 tcctgctgga gttcctgacc gccgccggga tcactgaggg catgaacgag ctgtacctcg
      721 agaagttttc tcaagaacag atcggcgaaa acattgtgtg cagggtcatt tgtaccacgg
      781 gtcaaattcc catccgagat ttgtcagctg atatttcaca agtgcttaag gaaaaacgat
      841 ccataaagaa agtttggaca tttggtagaa acccagcctg tgactatcat ttaggaaaca
      901 tttcaagact gtcaaataag catttccaaa tactactagg agaagacggt aaccttttat
      961 tgaatgacat ttccactaat gggacctggt taaatgggca aaaagtcgag aagaacagca
     1021 atcagttact gtcccaaggt gatgaaataa ccgttggtgt aggcgtggaa tcagatattt
     1081 tatctctggt cattttcata aacgacaaat ttaagcagtg cctggagcag aacaaagttg
     1141 atcgcggtac cagtgctggt ggtagtgctg gtggtagtgc tggtggtagt gctggtggta
     1201 gtgctggtgg ttccggcagt gctggtggta gtgctggtgg tagtaccagt gctggtggta
     1261 gtgctggtgg tagtgctggt ggtagtgctg gtggtagtgc tggtggttcc ggcagtgctg
     1321 gtggtagtgc tggtggtagt accagtgctg gtggtagtgc tggtggtagt gctggtggta
     1381 gtgctggtgg tagtgctggt ggttccggca gtgctggtgg tagtgctggt ggtagtacca
     1441 gtgctggtgg tagtgctggt ggtagtgctg gtggtagtgc tggtggtagt gctggtggtt
     1501 ccggagacag tgtcaagacc cccgaggatg aaggcaaccc actgctagag cagcttgaga
     1561 agaagggcgg ccgcatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     1621 tggtcgagct ggacggcgac gtcaacggcc acaagttcag cgtgtccggc gagggcgagg
     1681 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     1741 tgccctggcc caccctcgtg accaccctgt cctggggcgt gcagtgcttc gcccgctacc
     1801 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     1861 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     1921 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     1981 acatcctggg gcacaagctg gagtacaact acttcagcgg gaacgtctat atcaccgccg
     2041 acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggcg
     2101 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     2161 tgcccgacaa ccactacctg agcacccagt ccgccttaag caaagacccc aacgagaagc
     2221 gcgatcacat ggtcctgctg gagttcttga ccgccgccgg gggtaccaag gtgtacgacc
     2281 ccgagcagag gaagaggatg atcaccggcc cccagtggtg ggccaggtgc aagcagatga
     2341 acgtgctgga cagcttcatc aactactacg acagcgagaa gcacgccgag aacgccgtga
     2401 tcttcctgca cggcaacgcc actagcagct acctgtggag gcacgtggtg ccccacatcg
     2461 agcccgtggc caggtgcatc atccccgatc tgatcggcat gggcaagagc ggcaagagcg
     2521 gcaacggcag ctacaggctg ctggaccact acaagtacct gaccgcctgg ttcgagcttc
     2581 tgaacctgcc caagaagatc atcttcgtgg gccacgactg gggcgccgcc ctggccttcc
     2641 actacgccta cgagcaccag gacaggatca aggccatcgt gcacatggag agcgtggtgg
     2701 acgtgatcga gagctgggac gagtggccag acatcgagga ggacatcgcc ctgatcaaga
     2761 gcgaggaggg cgagaagatg gtgctggaga acaacttctt cgtggagacc gtgctgccca
     2821 gcaagatcat gagaaagctg gagcccgagg agttcgccgc ctacctggag cccttcaagg
     2881 agaagggcga ggtgagaaga cccaccctga gctggcccag agagatcccc ctggtgaagg
     2941 gcggcaagcc cgacgtggtg cagatcgtga gaaactacaa cgcctacctg agagccagcg
     3001 acgacctgcc caagctgttc atcgagggcg accccggctt cttcagcaac gccatcgtgg
     3061 agggcgccaa gaagttcccc aacaccgagt tcgtgaaggt gaagggcctg cacttcctcc
     3121 aggaggacgc ccccgacgag atgggcaagt acatcaagag cttcgtggag agagtgctga
     3181 agaacgagca gtctagactg cagctgcctc ctctggaacg cctgactctg gattaagtcg
     3241 acggatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg
     3301 acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt
     3361 ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg
     3421 gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa
     3481 agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc
     3541 ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc
     3601 ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca
     3661 gtcatagctg tccctcttct cttatggaga tccctcgacc tgcagcccaa gcttggcgta
     3721 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     3781 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     3841 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg
     3901 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3961 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4021 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4081 ctaggctttt gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca
     4141 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4201 ccaaactcat caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg
     4261 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4321 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4381 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4441 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4501 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4561 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4621 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4681 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4741 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4801 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4861 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4921 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4981 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5041 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5101 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5161 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5221 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5281 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5341 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5401 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5461 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5521 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5581 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5641 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5701 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5761 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5821 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5881 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5941 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6001 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6061 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6121 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6181 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgggtcga tcgacattga
     6241 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     6301 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     6361 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     6421 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     6481 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     6541 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     6601 gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc
     6661 tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg
     6721 ggggggggg
//
