LOCUS       sequence-419279-        1794 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Endogenously tag mouse EEA1 with an N-terminal 3xFLAG in addition to
            cytosolic mNG for cell sorting..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1794)
  AUTHORS   Hollingsworth LR, Veeraraghavan P, Paulo JA, Harper JW
  TITLE     Spatiotemporal proteomic profiling of cellular responses to NLRP3 
            agonists
  JOURNAL   bioRxiv 2024.04.19.590338
REFERENCE   2  (bases 1 to 1794)
  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: "bioRxiv 
            2024.04.19.590338"
COMMENT     Sequence Label: mNeonGreen-P2A-3xFLAG-msEEA1 HDR template
FEATURES             Location/Qualifiers
     source          1..1794
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             451..1158
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      706..715
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1186..1242
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1243..1308
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
ORIGIN
        1 caagctctcg gcggaagaag ccaaggcacc agcttcctgt ggcctgtctc tcacactagc
       61 cccgcttccg attggtcgaa gcctcgggcc ggccctccct ccggccccgc cctccccgcg
      121 gatggacagg ccgaggaggc ggggctgcag cggctccgcc cctcggcccc gggcgggcgg
      181 gcccgcccag ctgtcagtta cgggggcggc ccggcaggcg ggcgctcgct ggctccctcc
      241 tcgctggctc gtgctcgtga gcctctcgcc gcctgtgggg aagtcggggc cgccaaagcc
      301 gcgtccaccg ccgcctcttc cccgccctcc tccctcccgg ctcggctgcc ggcccctcgc
      361 gccctccagt cgtgagtaac ccgcataagg gcgcggagag tccccgcggc ggcggcggcg
      421 gtgacggcgg cggcggcggc cggccaaacc atggtgagca agggcgagga ggataacatg
      481 gcctctctcc cagcgacaca tgagttacac atctttggct ccatcaacgg tgtggacttt
      541 gacatggtgg gtcagggcac cggcaatcca aatgatggtt atgaggagtt aaacctgaag
      601 tccaccaagg gtgacctcca gttctccccc tggattctgg tccctcatat cgggtatggc
      661 ttccatcagt acctgcccta ccctgacggg atgtcgcctt tccaggccgc catggtagat
      721 ggctccggat accaagtcca tcgcacaatg cagtttgaag atggtgcctc ccttactgtt
      781 aactaccgct acacctacga gggaagccac atcaaaggag aggcccaggt gaaggggact
      841 ggtttccctg ctgacggtcc tgtgatgacc aactcgctga ccgctgcgga ctggtgcagg
      901 tcgaagaaga cttaccccaa cgacaaaacc atcatcagta cctttaagtg gagttacacc
      961 actggaaatg gcaagcgcta ccggagcact gcgcggacca cctacacctt tgccaagcca
     1021 atggcggcta actatctgaa gaaccagccg atgtacgtgt tccgtaagac ggagctcaag
     1081 cactccaaga ccgagctcaa cttcaaggag tggcaaaagg cctttaccga tgtgatgggc
     1141 atggacgagc tgtacaagtc tagaagcgct ggtaccggat ccggcgcaac aaacttctct
     1201 ctgctgaaac aagccggaga tgtcgaagag aatcctggac cggactacaa agaccatgac
     1261 ggtgattata aagatcatga catcgattac aaggatgacg atgacaaggg cggatccggg
     1321 ggaagcggcg gatcctttcg aagaatcttg cagagggtaa gagtgaaacc gtccttccgg
     1381 cagcacgtga gcgcgagcgt ccgagcccgg ccgcggggca gcccgggctg gagggcgggg
     1441 gcgctcagag ctccgctagg cccggtgaca gcgtgcgccg aggcctgctg gtcctgccgg
     1501 agccgggctg cgggtggggc ggggcgcagg ccggagcccg gagggcctcc agcctcggct
     1561 cccgcgctcg gggctgtcac cacgggctgg agggtgtggt gcccgcgcgc gcccctcccc
     1621 gctccccgct cccggctccg cgcccgtggg tgctcggctg cgggcggctg cggggccctt
     1681 ccctgcccca gcgtgccacc accttcccac gacacgcacc tgcctgtctg cctccttact
     1741 catctatctg cacggtacag gagggtcatg ggagacccgg cctagtcctg ggac
//
