LOCUS       sequence-101680-         641 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  For imaging of ERK kinase activity using a single polypeptide RA-WW 
            domain-ERK substrate-B protein and free GA-NES..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 641)
  AUTHORS   Ding Y, Li J, Enterina JR, Shen Y, Zhang I, Tewson PH, Mo GC, Zhang 
            J, Quinn AM, Hughes TE, Maysinger D, Alford SC, Zhang Y, Campbell RE
  TITLE     Ratiometric biosensors based on dimerization-dependent fluorescent 
            protein exchange.
  JOURNAL   Nat Methods. 2015 Jan 26. doi: 10.1038/nmeth.3261.
  PUBMED    25622108
REFERENCE   2  (bases 1 to 641)
  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: "Nat 
            Methods. 2015 Jan 26. doi: 10.1038/nmeth.3261."
COMMENT     Sequence Label: Ratiometric FPX sensor for ERK kinase activity
FEATURES             Location/Qualifiers
     source          1..641
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     37..56
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        37..55
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      75..84
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             96..113
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             117..149
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(153..161,162..176)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(316..334)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     584..603
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 actagagaac ccactgcctt actggcttat cgaaattaat acgactcact atagggagac
       61 ccaagcttgc ggccgccacc atggtgcggg gttctcatca tcatcatcat catggtatgg
      121 ctagcatgac tggtggacag caaatgggtc gggatctgta cgacgatgac gataaggatc
      181 ccatggtgag caagagcgag gaggtcatca aagagttcat gcgcttcaag gtgcgcttgg
      241 agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
      301 agggcaccca gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg
      361 acatcctgtc cccccagatc atgtacggct ccaaggcgta cgtgaagcac cccgccgacg
      421 tccccgatta catgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgcact
      481 tcgaggacgg cggtctggtg accgtgactc aggacacctc cctgcaggac ggcacgctga
      541 tctacaaggt gaagatgcgc ggcaccaact tcccccccga cggccccgta atgcagaaga
      601 agaccttggg ctgggattat gccaccgagc gcctgtaccc c
//
