LOCUS       sequence-273571-        6468 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Booster-ERK, a Red-Shifted Genetically Encoded FRET Biosensor for 
            ERK activity..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6468)
  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 6468)
  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-4511NES
FEATURES             Location/Qualifiers
     source          1..6468
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          596..1616
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1536..1558
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1624..1643
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1673..1682
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1679..2374
                     /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             2711..3364
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQFKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     CDS             3440..3466
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     complement(3482..3501)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3547..3602
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3601..3620)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3963..3979)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3963..3979)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3976..3998)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3987..4003
                     /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(4011..4017,4018..4035,4036..4041))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4056..4077
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4136..4331
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4182..4317
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4337..4471
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4374..4393)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4428..4447
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4539..4556)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4710..5298)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4790..4809)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5469..6260,6261..6329))
                     /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     6092..6111
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6330..6434)
                     /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 gggggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt
      541 tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt
      601 cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg
      661 gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg
      721 ctgtaattag cgcttggttt aatgacggct tgtttctttt ctgtggctgc gtgaaagcct
      781 tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg
      841 tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg
      901 gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg
      961 tgccccgcgg tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt
     1021 gggggggtga gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc
     1081 tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc
     1141 ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc
     1201 gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg ccggcggctg
     1261 tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg
     1321 acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1381 tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct
     1441 tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg
     1501 ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg
     1561 cggctctagc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc
     1621 tgggcaacgt gctggttatt gtgctgtctc atcattttgg caagaattcg ccgccaccat
     1681 ggtgagcgag ctgattaagg agaacatgca catgaagctg tacatggagg gcaccgtgaa
     1741 caaccaccac ttcaagtgca catccgaggg cgaaggcaag ccctacgagg gcacccagac
     1801 catgagaatc aaggcggtcg agggcggccc tctccccttc gccttcgaca tcctggctac
     1861 cagcttcatg tacggcagca aaaccttcat caaccacacc cagggcatcc ccgacttctt
     1921 taagcagtcc ttccccgagg gcttcacatg ggagagagtc accacatacg aagacggggg
     1981 cgtgctgacc gctacccagg acaccagcct ccaggacggc tgcctcatct acaacgtcaa
     2041 gatcagaggg gtgaacttcc catccaacgg ccctgtgatg cagaagaaaa cactcggctg
     2101 ggaggcctcc accgagaccc tgtaccccgc tgacggcggc ctggaaggca gagccgacat
     2161 ggccctgaag ctcgtgggcg ggggccacct gatctgcaac ttgaagacca catacagatc
     2221 caagaaaccc gctaagaacc tcaagatgcc cggcgtctac tatgtggaca gaagactgga
     2281 aagaatcaag gaggccgaca aagagaccta cgtcgagcag cacgaggtgg ctgtggccag
     2341 atactgcgac ctccctagca aactggggca cagactcgag atggcggacg aggagaagct
     2401 gccgcccggc tgggagaagc gcatgagccg cagctcaggc cgagtgtact acttcaacca
     2461 catcactaac gccagccagt gggagcggcc cagcggcaac agcagcagtg gtggcaaaaa
     2521 cgggcagggg gagcctgcca ggggtaccag tgctggtggt agtgctggtg gtagtgctgg
     2581 tggtagtgct ggtggtagtg ctggtggttc cggcagtgct ggtggtagtg ctggtggtag
     2641 taccagtgct ggtggtagtg ctggtggtag tgctggtggt agtgctggtg gtagtgctgg
     2701 tggttccgga atggtgagtg tgattaaacc agagatgaag atgaggtact acatggacgg
     2761 ctccgtcaat gggcatgagt tcacaattga aggtgaaggc acaggcagac cttacgaggg
     2821 acatcaagag atgacactac gcgtcacaat ggccgagggc gggccaatgc ctttcgcgtt
     2881 tgacttagtg tcacacgtgt tctgttacgg ccacagagta tttactaaat atccagaaga
     2941 gataccagac tatttcaaac aagcatttcc tgaaggcctg tcatgggaaa ggtcgttgga
     3001 gttcgaagat ggtgggtccg cttcagtcag tgcgcatata agccttagag gaaacacctt
     3061 ctaccacaaa tccaaattta ctggggttaa ctttcctgcc gatggtccta tcatgcaaaa
     3121 ccaaagtgtt gattgggagc catcaaccga gaaaattact gccagcgacg gagttctgaa
     3181 gggtgatgtt acgatgtacc taaaacttga aggaggcggc aatcacaaat gccaattcaa
     3241 gactacttac aaggcggcaa aagagattct tgaaatgcca ggagaccatt acatcggcca
     3301 tcgcctcgtc aggaaaaccg aaggcaacat tactgagcag gtagaagatg cagtagctca
     3361 ttccgcggcc gcaccagacg tccctagaac accggtggat aaagcaaagc tgtcattcca
     3421 atttccgtct agactgcagc tgcctcctct ggaacgcctg actctggatt aagtcgacgg
     3481 atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt
     3541 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct
     3601 cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt
     3661 agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag
     3721 gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga
     3781 cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa
     3841 aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca
     3901 tagctgtccc tcttctctta tggagatccc tcgacctgca gcccaagctt ggcgtaatca
     3961 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     4021 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4081 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc
     4141 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4201 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4261 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4321 gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4381 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4441 actcatcaat gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg
     4501 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4561 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4621 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4681 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4741 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4801 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4861 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4921 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4981 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5041 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5101 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     5161 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5221 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5281 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5341 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5401 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5461 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5521 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5581 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5641 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5701 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5761 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5821 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5881 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5941 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6001 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6061 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6121 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6181 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6241 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6301 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6361 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6421 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg ggtcgatc
//
