LOCUS       sequence-271043-        7698 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An ERK 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 7698)
  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 7698)
  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-6011nes
FEATURES             Location/Qualifiers
     source          1..7698
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        9..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..665
                     /label=chicken beta-actin promoter
     intron          666..1686
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1606..1628
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1694..1713
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(1744..1746,1747..1749,1750..2457)
                     /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             2632..2979
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             4678..4704
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     complement(4720..4739)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4785..4840
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4839..4858)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5201..5217)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5201..5217)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5214..5236)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5225..5241
                     /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(5249..5255,5256..5273,5274..5279))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5294..5315
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5374..5569
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5420..5555
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5575..5709
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5612..5631)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5666..5685
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5777..5794)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5948..6536)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6028..6047)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6707..7498,7499..7567))
                     /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     7330..7349
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7568..7672)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgatcga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
       61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      181 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      301 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      361 ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct
      421 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
      481 tgcagcgatg ggggcggggg gggggggggg gcgcgccagg cggggcgggg cggggcgagg
      541 ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa
      601 agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc
      661 gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc
      721 gcccgccccg gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt
      781 ctcctccggg ctgtaattag cgcttggttt aatgacggct tgtttctttt ctgtggctgc
      841 gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg
      901 cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg
      961 agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg
     1021 ccgggggcgg tgccccgcgg tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg
     1081 gtgtgtgcgt gggggggtga gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc
     1141 tgcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg
     1201 ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg
     1261 gggcggggcc gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg
     1321 ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga
     1381 gggcgcaggg acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc
     1441 gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg
     1501 gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct
     1561 gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
     1621 gtgtgaccgg cggctctagc tagagcctct gctaaccatg ttcatgcctt cttctttttc
     1681 ctacagctcc tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattc
     1741 ggcatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     1801 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     1861 tacggcaagc tgaccctgaa gcttctatgc accaccggca agctgcccgt gccctggccc
     1921 accctcgtga ccaccctggg ctacggcctg cagtgcttcg cccgctaccc cgaccacatg
     1981 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     2041 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     2101 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     2161 cacaagctgg agtacaacta caacagccac aacgtctata tcaccgccga caagcagaag
     2221 aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc
     2281 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     2341 cactacctga gctaccagtc cgccctgttc aaagacccca acgagaagcg cgatcacatg
     2401 gtcctgctgg agttcctgac cgccgccggg atcactgagg gcatgaacga gctgtacctc
     2461 gagatggcgg acgaggagaa gctgccgccc ggctgggaga agcgcatgag ccgcagctca
     2521 ggccgagtgt actacttcaa ccacatcact aacgccagcc agtgggagcg gcccagcggc
     2581 aacagcagca gtggtggcaa aaacgggcag ggggagcctg ccaggggtac cagtgctggt
     2641 ggtagtgctg gtggtagtgc tggtggtagt gctggtggta gtgctggtgg ttccggcagt
     2701 gctggtggta gtgctggtgg tagtaccagt gctggtggta gtgctggtgg tagtgctggt
     2761 ggtagtgctg gtggtagtgc tggtggttcc ggcagtgctg gtggtagtgc tggtggtagt
     2821 accagtgctg gtggtagtgc tggtggtagt gctggtggta gtgctggtgg tagtgctggt
     2881 ggttccggca gtgctggtgg tagtgctggt ggtagtacca gtgctggtgg tagtgctggt
     2941 ggtagtgctg gtggtagtgc tggtggtagt gctggtggtt ccggaccaga tgtccctaga
     3001 actccagtgg ataaagcaaa gctgtcattc caatttccgg gcggccgcat ggtgagcaag
     3061 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtcaac
     3121 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     3181 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     3241 ctgtcctggg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
     3301 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     3361 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     3421 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     3481 aactacttca gcgggaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc
     3541 aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccga ccactaccag
     3601 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     3661 cagtccgcct taagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     3721 ttgaccgccg ccgggggtac caaggtgtac gaccccgagc agaggaagag gatgatcacc
     3781 ggcccccagt ggtgggccag gtgcaagcag atgaacgtgc tggacagctt catcaactac
     3841 tacgacagcg agaagcacgc cgagaacgcc gtgatcttcc tgcacggcaa cgccactagc
     3901 agctacctgt ggaggcacgt ggtgccccac atcgagcccg tggccaggtg catcatcccc
     3961 gatctgatcg gcatgggcaa gagcggcaag agcggcaacg gcagctacag gctgctggac
     4021 cactacaagt acctgaccgc ctggttcgag cttctgaacc tgcccaagaa gatcatcttc
     4081 gtgggccacg actggggcgc cgccctggcc ttccactacg cctacgagca ccaggacagg
     4141 atcaaggcca tcgtgcacat ggagagcgtg gtggacgtga tcgagagctg ggacgagtgg
     4201 ccagacatcg aggaggacat cgccctgatc aagagcgagg agggcgagaa gatggtgctg
     4261 gagaacaact tcttcgtgga gaccgtgctg cccagcaaga tcatgagaaa gctggagccc
     4321 gaggagttcg ccgcctacct ggagcccttc aaggagaagg gcgaggtgag aagacccacc
     4381 ctgagctggc ccagagagat ccccctggtg aagggcggca agcccgacgt ggtgcagatc
     4441 gtgagaaact acaacgccta cctgagagcc agcgacgacc tgcccaagct gttcatcgag
     4501 ggcgaccccg gcttcttcag caacgccatc gtggagggcg ccaagaagtt ccccaacacc
     4561 gagttcgtga aggtgaaggg cctgcacttc ctccaggagg acgcccccga cgagatgggc
     4621 aagtacatca agagcttcgt ggagagagtg ctgaagaacg agcagtctag actgcagctg
     4681 cctcctctgg aacgcctgac tctggattaa gtcgacggat ctttttccct ctgccaaaaa
     4741 ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat
     4801 tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag
     4861 ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca
     4921 tatgctggct gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc
     4981 ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat
     5041 attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact
     5101 agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg
     5161 gagatccctc gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5221 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5281 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5341 tccagtcggg aaacctgtcg tgccagcgga tccgcatctc aattagtcag caaccatagt
     5401 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     5461 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     5521 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctaacttg
     5581 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5641 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5701 gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5761 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5821 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5881 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5941 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6001 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6061 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6121 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6181 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6241 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6301 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6361 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     6421 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6481 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6541 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6601 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6661 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6721 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6781 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6841 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6901 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6961 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7021 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7081 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7141 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7201 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7261 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7321 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7381 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7441 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7501 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7561 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7621 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7681 cccgaaaagt gccacctg
//
