LOCUS       sequence-267218-        7363 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Booster-JNK, a Red-Shifted Genetically Encoded FRET Biosensor for 
            JNK activity..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7363)
  AUTHORS   Watabe T, Terai K, Sumiyama K, Matsuda M
  TITLE     Booster, a Red-Shifted Genetically Encoded Forster Resonance Energy 
            Transfer (FRET) Biosensor Compatible with Cyan Fluorescent 
            Protein/Yellow Fluorescent Protein-Based FRET Biosensors and Blue 
            Light-Responsive Optogenetic Tools.
  JOURNAL   ACS Sens. 2020 Feb 26. doi: 10.1021/acssensors.9b01941.
  PUBMED    32101394
REFERENCE   2  (bases 1 to 7363)
  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: "ACS Sens. 
            2020 Feb 26. doi: 10.1021/acssensors.9b01941."
COMMENT     Sequence Label: pCAGGS-4580NES
FEATURES             Location/Qualifiers
     source          1..7363
                     /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"
     regulatory      1735..1744
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1741..2436
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     CDS             2872..3219
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             2968..3315
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             3064..3411
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             3160..3507
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             3256..3603
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             3610..4263
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQFKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     CDS             4345..4371
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     complement(4387..4406)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4452..4507
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4506..4525)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4866..4882)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4866..4882)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4879..4901)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4890..4906
                     /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(4914..4920,4921..4938,4939..4944))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4959..4980
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5039..5234
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5085..5220
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5240..5374
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5277..5296)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5331..5350
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5442..5459)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5613..6201)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5693..5712)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6372..7163,7164..7232))
                     /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     6995..7014
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7233..7337)
                     /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 cgccgccacc
     1741 atggtgagcg agctgattaa ggagaacatg cacatgaagc tgtacatgga gggcaccgtg
     1801 aacaaccacc acttcaagtg cacatccgag ggcgaaggca agccctacga gggcacccag
     1861 accatgagaa tcaaggcggt cgagggcggc cctctcccct tcgccttcga catcctggct
     1921 accagcttca tgtacggcag caaaaccttc atcaaccaca cccagggcat ccccgacttc
     1981 tttaagcagt ccttccccga gggcttcaca tgggagagag tcaccacata cgaagacggg
     2041 ggcgtgctga ccgctaccca ggacaccagc ctccaggacg gctgcctcat ctacaacgtc
     2101 aagatcagag gggtgaactt cccatccaac ggccctgtga tgcagaagaa aacactcggc
     2161 tgggaggcct ccaccgagac cctgtacccc gctgacggcg gcctggaagg cagagccgac
     2221 atggccctga agctcgtggg cgggggccac ctgatctgca acttgaagac cacatacaga
     2281 tccaagaaac ccgctaagaa cctcaagatg cccggcgtct actatgtgga cagaagactg
     2341 gaaagaatca aggaggccga caaagagacc tacgtcgagc agcacgaggt ggctgtggcc
     2401 agatactgcg acctccctag caaactgggg cacagactcg agaagttttc tcaagaacag
     2461 atcggcgaaa acattgtgtg cagggtcatt tgtaccacgg gtcaaattcc catccgagat
     2521 ttgtcagctg atatttcaca agtgcttaag gaaaaacgat ccataaagaa agtttggaca
     2581 tttggtagaa acccagcctg tgactatcat ttaggaaaca tttcaagact gtcaaataag
     2641 catttccaaa tactactagg agaagacggt aaccttttat tgaatgacat ttccactaat
     2701 gggacctggt taaatgggca aaaagtcgag aagaacagca atcagttact gtcccaaggt
     2761 gatgaaataa ccgttggtgt aggcgtggaa tcagatattt tatctctggt cattttcata
     2821 aacgacaaat ttaagcagtg cctggagcag aacaaagttg atcgcggtac cagtgctggt
     2881 ggtagtgctg gtggtagtgc tggtggtagt gctggtggta gtgctggtgg ttccggcagt
     2941 gctggtggta gtgctggtgg tagtaccagt gctggtggta gtgctggtgg tagtgctggt
     3001 ggtagtgctg gtggtagtgc tggtggttcc ggcagtgctg gtggtagtgc tggtggtagt
     3061 accagtgctg gtggtagtgc tggtggtagt gctggtggta gtgctggtgg tagtgctggt
     3121 ggttccggca gtgctggtgg tagtgctggt ggtagtacca gtgctggtgg tagtgctggt
     3181 ggtagtgctg gtggtagtgc tggtggtagt gctggtggtt ccggcagtgc tggtggtagt
     3241 gctggtggta gtaccagtgc tggtggtagt gctggtggta gtgctggtgg tagtgctggt
     3301 ggtagtgctg gtggttccgg cagtgctggt ggtagtgctg gtggtagtac cagtgctggt
     3361 ggtagtgctg gtggtagtgc tggtggtagt gctggtggta gtgctggtgg ttccggcagt
     3421 gctggtggta gtgctggtgg tagtaccagt gctggtggta gtgctggtgg tagtgctggt
     3481 ggtagtgctg gtggtagtgc tggtggttcc ggcagtgctg gtggtagtgc tggtggtagt
     3541 accagtgctg gtggtagtgc tggtggtagt gctggtggta gtgctggtgg tagtgctggt
     3601 ggttccggaa tggtgagtgt gattaaacca gagatgaaga tgaggtacta catggacggc
     3661 tccgtcaatg ggcatgagtt cacaattgaa ggtgaaggca caggcagacc ttacgaggga
     3721 catcaagaga tgacactacg cgtcacaatg gccgagggcg ggccaatgcc tttcgcgttt
     3781 gacttagtgt cacacgtgtt ctgttacggc cacagagtat ttactaaata tccagaagag
     3841 ataccagact atttcaaaca agcatttcct gaaggcctgt catgggaaag gtcgttggag
     3901 ttcgaagatg gtgggtccgc ttcagtcagt gcgcatataa gccttagagg aaacaccttc
     3961 taccacaaat ccaaatttac tggggttaac tttcctgccg atggtcctat catgcaaaac
     4021 caaagtgttg attgggagcc atcaaccgag aaaattactg ccagcgacgg agttctgaag
     4081 ggtgatgtta cgatgtacct aaaacttgaa ggaggcggca atcacaaatg ccaattcaag
     4141 actacttaca aggcggcaaa agagattctt gaaatgccag gagaccatta catcggccat
     4201 cgcctcgtca ggaaaaccga aggcaacatt actgagcagg tagaagatgc agtagctcat
     4261 tccgcggccg cagacagtgt caagaccccc gaggatgaag gcaacccact gctagagcaa
     4321 ttggagaaga agtctagact gcagctgcct cctctggaac gcctgactct ggattaagtc
     4381 gacggatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct
     4441 gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg
     4501 tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt
     4561 ggtttagagt ttggcaacat atgccatatg ctggctgcca tgaacaaagg tggctataaa
     4621 gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct
     4681 tgacttgagg ttagattttt ttttatattt tgtttgtgtt atttttttct ttaacatccc
     4741 taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag
     4801 tcatagctgt ccctcttctc ttatgaagat ccctcgacct gcagcccaag cttggcgtaa
     4861 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4921 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4981 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc
     5041 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     5101 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     5161 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     5221 taggcttttg caaaaagcta acttgtttat tgcagcttat aatggttaca aataaagcaa
     5281 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5341 caaactcatc aatgtatctt atcatgtctg gatccgctgc attaatgaat cggccaacgc
     5401 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5461 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5521 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5581 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5641 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5701 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5761 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt
     5821 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5881 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5941 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6001 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     6061 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6121 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6181 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6241 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6301 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6361 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6421 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6481 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6541 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6601 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6661 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6721 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6781 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6841 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6901 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6961 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7021 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7081 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7141 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7201 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7261 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7321 caaatagggg ttccgcgcac atttccccga aaagtgccac ctg
//
