LOCUS       sequence-325716-        2266 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Optimized EKAREV FRET biosensor sensor for ERK.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2266)
  AUTHORS   Ponsioen B, Post JB, Buissant des Amorie JR, Laskaris D, van Ineveld
            RL, Kersten S, Bertotti A, Sassi F, Sipieter F, Cappe B, Mertens S, 
            Verlaan-Klink I, Boj SF, Vries RGJ, Rehmann H, Vandenabeele P, 
            Riquet FB, Trusolino L, Bos JL, Snippert HJG
  TITLE     Quantifying single-cell ERK dynamics in colorectal cancer organoids 
            reveals EGFR as an amplifier of oncogenic MAPK pathway signalling.
  JOURNAL   Nat Cell Biol. 2021 Apr;23(4):377-390. doi: 
            10.1038/s41556-021-00654-5. Epub 2021 Apr 1.
  PUBMED    33795873
REFERENCE   2  (bases 1 to 2266)
  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"; doi: 
            "10.1038/s41556-021-00654-5"; journalName: "Nat Cell Biol"; date: 
            "2021-04"; volume: "23"; issue: "4"; pages: "377-390"
COMMENT     Sequence Label: miniTol2 x EF1a_EKAREN4 + PGK-puro
FEATURES             Location/Qualifiers
     source          1..2266
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             join(1..3,4..6,7..714)
                     /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"
     primer_bind     complement(307..326)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             889..1236
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             2023..2043
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2121..2255
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2145..2164)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2199..2218
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
       61 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      121 ggcaagctga ccctgaagct tctatgcacc accggcaagc tgcccgtgcc ctggcccacc
      181 ctcgtgacca ccctgggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag
      241 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      301 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      361 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      421 aagctggagt acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac
      481 ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc
      541 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
      601 tacctgagct accagtccgc cctgttcaaa gaccccaacg agaagcgcga tcacatggtc
      661 ctgctggagt tcctgaccgc cgccgggatc actgagggca tgaacgagct gtacctcgag
      721 atggcggacg aggagaagct gccgcccggc tgggagaagc gcatgagccg cagctcaggc
      781 cgagtgtact acttcaacca catcactaac gccagccagt gggagcggcc cagcggcaac
      841 agcagcagtg gtggcaaaaa cgggcagggg gagcctgcca ggggtaccag tgctggtggt
      901 agtgctggtg gtagtgctgg tggtagtgct ggtggtagtg ctggtggttc cggcagtgct
      961 ggtggtagtg ctggtggtag taccagtgct ggtggtagtg ctggtggtag tgctggtggt
     1021 agtgctggtg gtagtgctgg tggttccggc agtgctggtg gtagtgctgg tggtagtacc
     1081 agtgctggtg gtagtgctgg tggtagtgct ggtggtagtg ctggtggtag tgctggtggt
     1141 tccggcagtg ctggtggtag tgctggtggt agtaccagtg ctggtggtag tgctggtggt
     1201 agtgctggtg gtagtgctgg tggtagtgct ggtggttccg gaccagatgt ccctagaact
     1261 ccagtggatc cagcatggct gtcattccaa tttccgggcg gccgcatggt gagtaaaggg
     1321 gaagaactgt ttactggcgt ggtgccaatt ctggtcgaac tggatggaga tgtaaatggg
     1381 cataaattta gtgtgtctgg agaaggggaa ggagacgcta catatgggaa actgacactg
     1441 aaatttattt gtactactgg aaaactgcct gtgccttggc ctacactcgt gacaaccctg
     1501 tcttggggag tgcagtgttt tgcccggtat cctgatcata tgaaacagca tgattttttc
     1561 aaatctgcca tgcctgaggg atatgtccag gaacggacaa ttttctttaa agatgatggg
     1621 aattataaaa cacgggccga agtgaaattt gaaggggata cactggtgaa tcggattgaa
     1681 ctgaaaggaa ttgattttaa agaagatggg aatattctgg gccataaact ggaatataat
     1741 tatttcagcg ataatgtcta cattacagcc gataaacaga agaatgggat taaagccaat
     1801 tttaaaattc ggcataatat tgaagatgga ggggtgcagc tcgccgatca ttatcagcag
     1861 aatacaccta ttggggatgg gcctgtgctg ctgcctgata atcattatct gagcacacag
     1921 tctgccctga gcaaggaccc taatgaaaag cgggaccata tggtcctgct ggaatttctg
     1981 actgccgccg gcattacact cgggatggat gaactgtcta gacctaagaa gaagcgtaag
     2041 gtgtaagtcg acggtcgagt cgagacgcta gccaccggtc gttaacatta agctagaact
     2101 atagtgagtc gtattacgta gatccagaca tgataagata cattgatgag tttggacaaa
     2161 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     2221 tatttgtaac cattataagc tgcaataaac aagttaacaa cgccgg
//
