LOCUS       sequence-204832-        7687 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 7687)
  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 7687)
  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..7687
                     /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             2623..2970
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             4669..4695
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     complement(4711..4730)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4776..4831
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4830..4849)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5190..5206)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5190..5206)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5203..5225)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5214..5230
                     /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(5238..5244,5245..5262,5263..5268))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5283..5304
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5363..5558
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5409..5544
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5564..5698
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5601..5620)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5655..5674
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5766..5783)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5937..6525)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6017..6036)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6696..7487,7488..7556))
                     /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     7319..7338
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7557..7661)
                     /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 cgagatggcg
     2461 gacgaggaga agctgccgcc cggctgggag aagcgcatga gccgcagctc aggccgagtg
     2521 tactacttca accacatcac taacgccagc cagtgggagc ggcccagcgg caacagcagc
     2581 agtggtggca aaaacgggca gggggagcct gccaggggta ccagtgctgg tggtagtgct
     2641 ggtggtagtg ctggtggtag tgctggtggt agtgctggtg gttccggcag tgctggtggt
     2701 agtgctggtg gtagtaccag tgctggtggt agtgctggtg gtagtgctgg tggtagtgct
     2761 ggtggtagtg ctggtggttc cggcagtgct ggtggtagtg ctggtggtag taccagtgct
     2821 ggtggtagtg ctggtggtag tgctggtggt agtgctggtg gtagtgctgg tggttccggc
     2881 agtgctggtg gtagtgctgg tggtagtacc agtgctggtg gtagtgctgg tggtagtgct
     2941 ggtggtagtg ctggtggtag tgctggtggt tccggaccag atgtccctag aactccagtg
     3001 gataaagcaa agctgtcatt ccaatttccg ggcggccgca tggtgagcaa gggcgaggag
     3061 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtcaa cggccacaag
     3121 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     3181 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgtcctgg
     3241 ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     3301 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     3361 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     3421 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacttc
     3481 agcgggaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag
     3541 atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca gcagaacacc
     3601 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     3661 ttaagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cttgaccgcc
     3721 gccgggggta ccaaggtgta cgaccccgag cagaggaaga ggatgatcac cggcccccag
     3781 tggtgggcca ggtgcaagca gatgaacgtg ctggacagct tcatcaacta ctacgacagc
     3841 gagaagcacg ccgagaacgc cgtgatcttc ctgcacggca acgccactag cagctacctg
     3901 tggaggcacg tggtgcccca catcgagccc gtggccaggt gcatcatccc cgatctgatc
     3961 ggcatgggca agagcggcaa gagcggcaac ggcagctaca ggctgctgga ccactacaag
     4021 tacctgaccg cctggttcga gcttctgaac ctgcccaaga agatcatctt cgtgggccac
     4081 gactggggcg ccgccctggc cttccactac gcctacgagc accaggacag gatcaaggcc
     4141 atcgtgcaca tggagagcgt ggtggacgtg atcgagagct gggacgagtg gccagacatc
     4201 gaggaggaca tcgccctgat caagagcgag gagggcgaga agatggtgct ggagaacaac
     4261 ttcttcgtgg agaccgtgct gcccagcaag atcatgagaa agctggagcc cgaggagttc
     4321 gccgcctacc tggagccctt caaggagaag ggcgaggtga gaagacccac cctgagctgg
     4381 cccagagaga tccccctggt gaagggcggc aagcccgacg tggtgcagat cgtgagaaac
     4441 tacaacgcct acctgagagc cagcgacgac ctgcccaagc tgttcatcga gggcgacccc
     4501 ggcttcttca gcaacgccat cgtggagggc gccaagaagt tccccaacac cgagttcgtg
     4561 aaggtgaagg gcctgcactt cctccaggag gacgcccccg acgagatggg caagtacatc
     4621 aagagcttcg tggagagagt gctgaagaac gagcagtcta gactgcagct gcctcctctg
     4681 gaacgcctga ctctggatta agtcgacgga tctttttccc tctgccaaaa attatgggga
     4741 catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc
     4801 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca
     4861 tttaaaacat cagaatgagt atttggttta gagtttggca acatatgcca tatgctggct
     4921 gccatgaaca aaggtggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc
     4981 attccttatt ccatagaaaa gccttgactt gaggttagat ttttttttat attttgtttg
     5041 tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt
     5101 tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatga agatccctcg
     5161 acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     5221 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     5281 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5341 aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc ccgcccctaa
     5401 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     5461 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     5521 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc
     5581 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5641 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccg
     5701 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5761 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5821 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5881 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5941 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     6001 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6061 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6121 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6181 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6241 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6301 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6361 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6421 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6481 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6541 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6601 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6661 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6721 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6781 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6841 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6901 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6961 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     7021 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     7081 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     7141 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7201 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7261 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7321 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7381 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7441 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7501 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7561 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7621 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7681 ccacctg
//
