LOCUS       sequence-273572-        6748 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 6748)
  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 6748)
  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..6748
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          609..1629
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1549..1571
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1637..1656
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1686..1695
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1692..2387
                     /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             2985..3638
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQFKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     CDS             3720..3746
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     complement(3762..3781)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3827..3882
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3881..3900)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4243..4259)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4243..4259)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4256..4278)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4267..4283
                     /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(4291..4297,4298..4315,4316..4321))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4336..4357
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4416..4611
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4462..4597
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4617..4751
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4654..4673)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4708..4727
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4819..4836)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4990..5578)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5070..5089)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5749..6540,6541..6609))
                     /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     6372..6391
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6610..6714)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc
      541 gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc
      601 ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc
      661 gccgcccgcc ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc
      721 cttctcctcc gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc
      781 tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggg
      841 gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct
      901 gtgagcgctg cgggcgcggc gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg
      961 cggccggggg cggtgccccg cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc
     1021 ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc
     1081 ccctgcaccc ccctccccga gttgctgagc acggcccggc ttcgggtgcg gggctccgta
     1141 cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg
     1201 gcggggcggg gccgcctcgg gccggggagg gctcggggga ggggcgcggc ggcccccgga
     1261 gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg taatcgtgcg
     1321 agagggcgca gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc
     1381 gccgcacccc ctctagcggg cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg
     1441 gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct tctccctctc cagcctcggg
     1501 gctgtccgcg gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct
     1561 ggcgtgtgac cggcggctct agctagagcc tctgctaacc atgttcatgc cttcttcttt
     1621 ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaagaat
     1681 tcgccgccac catggtgagc gagctgatta aggagaacat gcacatgaag ctgtacatgg
     1741 agggcaccgt gaacaaccac cacttcaagt gcacatccga gggcgaaggc aagccctacg
     1801 agggcaccca gaccatgaga atcaaggcgg tcgagggcgg ccctctcccc ttcgccttcg
     1861 acatcctggc taccagcttc atgtacggca gcaaaacctt catcaaccac acccagggca
     1921 tccccgactt ctttaagcag tccttccccg agggcttcac atgggagaga gtcaccacat
     1981 acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccaggac ggctgcctca
     2041 tctacaacgt caagatcaga ggggtgaact tcccatccaa cggccctgtg atgcagaaga
     2101 aaacactcgg ctgggaggcc tccaccgaga ccctgtaccc cgctgacggc ggcctggaag
     2161 gcagagccga catggccctg aagctcgtgg gcgggggcca cctgatctgc aacttgaaga
     2221 ccacatacag atccaagaaa cccgctaaga acctcaagat gcccggcgtc tactatgtgg
     2281 acagaagact ggaaagaatc aaggaggccg acaaagagac ctacgtcgag cagcacgagg
     2341 tggctgtggc cagatactgc gacctcccta gcaaactggg gcacagactc gagaagtttt
     2401 ctcaagaaca gatcggcgaa aacattgtgt gcagggtcat ttgtaccacg ggtcaaattc
     2461 ccatccgaga tttgtcagct gatatttcac aagtgcttaa ggaaaaacga tccataaaga
     2521 aagtttggac atttggtaga aacccagcct gtgactatca tttaggaaac atttcaagac
     2581 tgtcaaataa gcatttccaa atactactag gagaagacgg taacctttta ttgaatgaca
     2641 tttccactaa tgggacctgg ttaaatgggc aaaaagtcga gaagaacagc aatcagttac
     2701 tgtcccaagg tgatgaaata accgttggtg taggcgtgga atcagatatt ttatctctgg
     2761 tcattttcat aaacgacaaa tttaagcagt gcctggagca gaacaaagtt gatcgcggta
     2821 ccagtgctgg tggtagtgct ggtggtagtg ctggtggtag tgctggtggt agtgctggtg
     2881 gttccggcag tgctggtggt agtgctggtg gtagtaccag tgctggtggt agtgctggtg
     2941 gtagtgctgg tggtagtgct ggtggtagtg ctggtggttc cggaatggtg agtgtgatta
     3001 aaccagagat gaagatgagg tactacatgg acggctccgt caatgggcat gagttcacaa
     3061 ttgaaggtga aggcacaggc agaccttacg agggacatca agagatgaca ctacgcgtca
     3121 caatggccga gggcgggcca atgcctttcg cgtttgactt agtgtcacac gtgttctgtt
     3181 acggccacag agtatttact aaatatccag aagagatacc agactatttc aaacaagcat
     3241 ttcctgaagg cctgtcatgg gaaaggtcgt tggagttcga agatggtggg tccgcttcag
     3301 tcagtgcgca tataagcctt agaggaaaca ccttctacca caaatccaaa tttactgggg
     3361 ttaactttcc tgccgatggt cctatcatgc aaaaccaaag tgttgattgg gagccatcaa
     3421 ccgagaaaat tactgccagc gacggagttc tgaagggtga tgttacgatg tacctaaaac
     3481 ttgaaggagg cggcaatcac aaatgccaat tcaagactac ttacaaggcg gcaaaagaga
     3541 ttcttgaaat gccaggagac cattacatcg gccatcgcct cgtcaggaaa accgaaggca
     3601 acattactga gcaggtagaa gatgcagtag ctcattccgc ggccgcagac agtgtcaaga
     3661 cccccgagga tgaaggcaac ccactgctag agcaattgga gaagaagtct agactgcagc
     3721 tgcctcctct ggaacgcctg actctggatt aagtcgacgg atctttttcc ctctgccaaa
     3781 aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt
     3841 attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg
     3901 agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc
     3961 catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag
     4021 ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta gatttttttt
     4081 atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta
     4141 ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta
     4201 tggagatccc tcgacctgca gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt
     4261 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4321 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4381 tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc agcaaccata
     4441 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4501 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     4561 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctaact
     4621 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4681 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     4741 atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4801 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4861 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4921 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4981 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5041 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5101 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5161 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5221 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5281 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5341 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5401 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     5461 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5521 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5581 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5641 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5701 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5761 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5821 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5881 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5941 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6001 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6061 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6121 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6181 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6241 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6301 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6361 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6421 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6481 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6541 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6601 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6661 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6721 tccccgaaaa gtgccacctg ggtcgatc
//
