LOCUS       sequence-204834-        7954 bp    DNA     circular SYN 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      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7954)
  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 7954)
  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..7954
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        8..387
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..665
                     /label=chicken beta-actin promoter
     intron          666..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1598..1620
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             join(1735..1737,1738..1740,1741..2448)
                     /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             2884..3231
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             4936..4962
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     complement(4978..4997)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5043..5098
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5097..5116)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5457..5473)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5457..5473)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5470..5492)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5481..5497
                     /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(5505..5511,5512..5529,5530..5535))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5550..5571
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5630..5825
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5676..5811
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5831..5965
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5868..5887)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5922..5941
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6033..6050)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6204..6792)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6284..6303)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6963..7754,7755..7823))
                     /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     7586..7605
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7824..7928)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgatcgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
       61 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      121 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      181 atagggactt tccattgacg tcaatgggtg gactatttac ggtaaactgc ccacttggca
      241 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      301 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      361 tacgtattag tcatcgctat taccatgggt cgaggtgagc cccacgttct gcttcactct
      421 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
      481 tgcagcgatg ggggcggggg gggggggggc gcgcgccagg cggggcgggg cgggggcgag
      541 ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa
      601 agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc
      661 gggcgggagt cgctgcgttg ccttcgcccc gtgccccgct ccgcgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggctcgt ttcttttctg tggctgcgtg
      841 aaagccttaa agggctccgg gagggccctt tgtgcggggg ggagcggctc ggggggtgcg
      901 tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggcccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcgtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
     1081 tgcgtggggg ggtgagcagg gggtgtgggc gcggcggtcg ggctgtaacc cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtgcggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccg gagcgccggc
     1321 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
     1381 cagggacttc ctttgtccca aatctggcgg agccgaaatc tgggaggcgc cgccgcaccc
     1441 cctctagcgg gcgcgggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg
     1501 ccttcgtgcg tcgccgcgcc gccgtcccct tctccatctc cagcctcggg gctgccgcag
     1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1621 ggcggctcta gctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt ggcaagaatt cggcatggtg
     1741 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     1801 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     1861 ctgaccctga agcttctatg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     1921 accaccctgg gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac
     1981 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     2041 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     2101 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     2161 gagtacaact acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc
     2221 aaggccaact tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac
     2281 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     2341 agctaccagt ccgccctgtt caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     2401 gagttcctga ccgccgccgg gatcactgag ggcatgaacg agctgtacct cgagaagttt
     2461 tctcaagaac agatcggcga aaacattgtg tgcagggtca tttgtaccac gggtcaaatt
     2521 cccatccgag atttgtcagc tgatatttca caagtgctta aggaaaaacg atccataaag
     2581 aaagtttgga catttggtag aaacccagcc tgtgactatc atttaggaaa catttcaaga
     2641 ctgtcaaata agcatttcca aatactacta ggagaagacg gtaacctttt attgaatgac
     2701 atttccacta atgggacctg gttaaatggg caaaaagtcg agaagaacag caatcagtta
     2761 ctgtcccaag gtgatgaaat aaccgttggt gtaggcgtgg aatcagatat tttatctctg
     2821 gtcattttca taaacgacaa atttaagcag tgcctggagc agaacaaagt tgatcgcggt
     2881 accagtgctg gtggtagtgc tggtggtagt gctggtggta gtgctggtgg tagtgctggt
     2941 ggttccggca gtgctggtgg tagtgctggt ggtagtacca gtgctggtgg tagtgctggt
     3001 ggtagtgctg gtggtagtgc tggtggtagt gctggtggtt ccggcagtgc tggtggtagt
     3061 gctggtggta gtaccagtgc tggtggtagt gctggtggta gtgctggtgg tagtgctggt
     3121 ggtagtgctg gtggttccgg cagtgctggt ggtagtgctg gtggtagtac cagtgctggt
     3181 ggtagtgctg gtggtagtgc tggtggtagt gctggtggta gtgctggtgg ttccggagac
     3241 agtgtcaaga cccccgagga tgaaggcaac ccactgctag agcagcttga gaagaagggc
     3301 ggccgcatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     3361 ctggacggcg acgtcaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     3421 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     3481 cccaccctcg tgaccaccct gtcctggggc gtgcagtgct tcgcccgcta ccccgaccac
     3541 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     3601 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     3661 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     3721 gggcacaagc tggagtacaa ctacttcagc gggaacgtct atatcaccgc cgacaagcag
     3781 aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag
     3841 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     3901 aaccactacc tgagcaccca gtccgcctta agcaaagacc ccaacgagaa gcgcgatcac
     3961 atggtcctgc tggagttctt gaccgccgcc gggggtacca aggtgtacga ccccgagcag
     4021 aggaagagga tgatcaccgg cccccagtgg tgggccaggt gcaagcagat gaacgtgctg
     4081 gacagcttca tcaactacta cgacagcgag aagcacgccg agaacgccgt gatcttcctg
     4141 cacggcaacg ccactagcag ctacctgtgg aggcacgtgg tgccccacat cgagcccgtg
     4201 gccaggtgca tcatccccga tctgatcggc atgggcaaga gcggcaagag cggcaacggc
     4261 agctacaggc tgctggacca ctacaagtac ctgaccgcct ggttcgagct tctgaacctg
     4321 cccaagaaga tcatcttcgt gggccacgac tggggcgccg ccctggcctt ccactacgcc
     4381 tacgagcacc aggacaggat caaggccatc gtgcacatgg agagcgtggt ggacgtgatc
     4441 gagagctggg acgagtggcc agacatcgag gaggacatcg ccctgatcaa gagcgaggag
     4501 ggcgagaaga tggtgctgga gaacaacttc ttcgtggaga ccgtgctgcc cagcaagatc
     4561 atgagaaagc tggagcccga ggagttcgcc gcctacctgg agcccttcaa ggagaagggc
     4621 gaggtgagaa gacccaccct gagctggccc agagagatcc ccctggtgaa gggcggcaag
     4681 cccgacgtgg tgcagatcgt gagaaactac aacgcctacc tgagagccag cgacgacctg
     4741 cccaagctgt tcatcgaggg cgaccccggc ttcttcagca acgccatcgt ggagggcgcc
     4801 aagaagttcc ccaacaccga gttcgtgaag gtgaagggcc tgcacttcct ccaggaggac
     4861 gcccccgacg agatgggcaa gtacatcaag agcttcgtgg agagagtgct gaagaacgag
     4921 cagtctagac tgcagctgcc tcctctggaa cgcctgactc tggattaagt cgacggatct
     4981 ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg
     5041 ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact
     5101 cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag
     5161 tttggcaaca tatgccatat gctggctgcc atgaacaaag gtggctataa agaggtcatc
     5221 agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag
     5281 gttagatttt tttttatatt ttgtttgtgt tatttttttc tttaacatcc ctaaaatttt
     5341 ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg
     5401 tccctcttct cttatgaaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca
     5461 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     5521 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     5581 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt
     5641 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     5701 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     5761 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     5821 gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     5881 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5941 caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga
     6001 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6061 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6121 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6181 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6241 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6301 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6361 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     6421 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6481 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6541 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6601 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     6661 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6721 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6781 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6841 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6901 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6961 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7021 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7081 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7141 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7201 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7261 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7321 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7381 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7441 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7501 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7561 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7621 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7681 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7741 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7801 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7861 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7921 gttccgcgca catttccccg aaaagtgcca cctg
//
