LOCUS       sequence-325703-        1384 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 1384)
  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 1384)
  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_EKAREN5 + PGK-puro
FEATURES             Location/Qualifiers
     source          1..1384
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        8..507
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(43..66)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     440..459
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     promoter        519..566
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             615..1214
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(615..634)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1111..1131
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    1243..1377
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1267..1286)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1321..1340
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 ccgcaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc
       61 ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc
      121 ggtaggcgcc aaccggctcc gttctttggt ggccccttcg cgccaccttc tactcctccc
      181 ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa
      241 gtagcacgtc tcactagtct cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta
      301 ggcctttggg gcagcggcca atagcagctt tgctccttcg ctttctgggc tcagaggctg
      361 ggaaggggtg ggtccggggg cgggctcagg ggcgggctca ggggcggggc gggcgcccga
      421 aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc
      481 tcctcttcct catctccggg cctttcgacc tgcagcctgt tgacaattaa tcatcggcat
      541 agtatatcgg catagtataa tacgacaagg tgaggaacta aaccgtcgac gatttcacta
      601 gtctcgaagc caccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
      661 tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
      721 ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
      781 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
      841 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
      901 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
      961 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg
     1021 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
     1081 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
     1141 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
     1201 agcccggtgc ctgactagaa ctatagtgag tcgtattacg tagatccaga catgataaga
     1261 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     1321 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     1381 aaca
//
