LOCUS       sequence-325717-        1393 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 1393)
  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 1393)
  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..1393
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        10..509
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(45..68)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     442..461
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     promoter        521..568
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             617..1216
                     /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(617..636)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1113..1133
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    1245..1379
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1269..1288)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1323..1342
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 ggccgcaccg ggtaggggag gcgcttttcc caaggcagtc tggagcatgc gctttagcag
       61 ccccgctggg cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca
      121 ccggtaggcg ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc
      181 ccctagtcag gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg
      241 aagtagcacg tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg
      301 taggcctttg gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc
      361 tgggaagggg tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc
      421 gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt
      481 tctcctcttc ctcatctccg ggcctttcga cctgcagcct gttgacaatt aatcatcggc
      541 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccgtcg acgatttcac
      601 tagtctcgaa gccaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
      661 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
      721 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
      781 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
      841 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
      901 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
      961 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     1021 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     1081 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     1141 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     1201 caagcccggt gcctgactag aactatagtg agtcgtatta cgtagatcca gacatgataa
     1261 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     1321 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     1381 acaacaacaa ttg
//
