LOCUS       sequence-419284-        5124 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Express the human EndoTag (3xFLAG-EEA1) for EndoIPs. mTagBFP2 to 
            sort for even expression levels..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5124)
  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 5124)
  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: pLV-UBC mTagBFP2-P2A-3xFLAG-hEEA1
FEATURES             Location/Qualifiers
     source          1..5124
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1..699
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             709..765
                     /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             769..792
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             796..819
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             823..846
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    862..886
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             892..5124
                     /codon_start=1
                     /product="human early endosome antigen 1"
                     /label=EEA1
                     /translation="MLRRILQRTPGRVGSQGSDLDSSATPINTVDVNNESSSEGFICPQ
                     CMKSLGSADELFKHYEAVHDAGNDSGHGGESNLALKRDDVTLLRQEVQDLQASLKEEKW
                     YSEELKKELEKYQGLQQQEAKPDGLVTDSSAELQSLEQQLEEAQTENFNIKQMKDLFEQ
                     KAAQLATEIADIKSKYDEERSLREAAEQKVTRLTEELNKEATVIQDLKTELLQRPGIED
                     VAVLKKELVQVQTLMDNMTLERERESEKLKDECKKLQSQYASSEATISQLRSELAKGPQ
                     EVAVYVQELQKLKSSVNELTQKNQTLTENLLKKEQDYTKLEEKHNEESVSKKNIQATLH
                     QKDLDCQQLQSRLSASETSLHRIHVELSEKGEATQKLKEELSEVETKYQHLKAEFKQLQ
                     QQREEKEQHGLQLQSEINQLHSKLLETERQLGEAHGRLKEQRQLSSEKLMDKEQQVADL
                     QLKLSRLEEQLKEKVTNSTELQHQLDKTKQQHQEQQALQQSTTAKLREAQNDLEQVLRQ
                     IGDKDQKIQNLEALLQKSKENISLLEKEREDLYAKIQAGEGETAVLNQLQEKNHTLQEQ
                     VTQLTEKLKNQSESHKQAQENLHDQVQEQKAHLRAAQDRVLSLETSVNELNSQLNESKE
                     KVSQLDIQIKAKTELLLSAEAAKTAQRADLQNHLDTAQNALQDKQQELNKITTQLDQVT
                     AKLQDKQEHCSQLESHLKEYKEKYLSLEQKTEELEGQIKKLEADSLEVKASKEQALQDL
                     QQQRQLNTDLELRATELSKQLEMEKEIVSSTRLDLQKKSEALESIKQKLTKQEEEKKIL
                     KQDFETLSQETKIQHEELNNRIQTTVTELQKVKMEKEALMTELSTVKDKLSKVSDSLKN
                     SKSEFEKENQKGKAAILDLEKTCKELKHQLQVQMENTLKEQKELKKSLEKEKEASHQLK
                     LELNSMQEQLIQAQNTLKQNEKEEQQLQGNINELKQSSEQKKKQIEALQGELKIAVLQK
                     TELENKLQQQLTQAAQELAAEKEKISVLQNNYEKSQETFKQLQSDFYGRESELLATRQD
                     LKSVEEKLSLAQEDLISNRNQIGNQNKLIQELKTAKATLEQDSAKKEQQLQERCKALQD
                     IQKEKSLKEKELVNEKSKLAEIEEIKCRQEKEITKLNEELKSHKLESIKEITNLKDAKQ
                     LLIQQKLELQGKADSLKAAVEQEKRNQQILKDQVKKEEEELKKEFIEKEAKLHSEIKEK
                     EVGMKKHEENEAKLTMQITALNENLGTVKKEWQSSQRRVSELEKQTDDLRGEIAVLEAT
                     VQNNQDERRALLERCLKGEGEIEKLQTKVLELQRKLDNTTAAVQELGRENQSLQIKHTQ
                     ALNRKWAEDNEVQNCMACGKGFSVTVRRHHCRQCGNIFCAECSAKNALTPSSKKPVRVC
                     DACFNDLQG"
ORIGIN
        1 atgagcgagc tgattaagga gaacatgcac atgaagctgt acatggaggg caccgtggac
       61 aaccatcact tcaagtgcac atccgagggc gaaggcaagc cctacgaggg cacccagacc
      121 atgagaatca aggtggtcga gggcggccct ctccccttcg ccttcgacat cctggctact
      181 agcttcctct acggcagcaa gaccttcatc aaccacaccc agggcatccc cgacttcttc
      241 aagcagtcct tccctgaggg cttcacatgg gagagagtca ccacatacga agacgggggc
      301 gtgctgaccg ctacccagga caccagcctc caggacggct gcctcatcta caacgtcaag
      361 atcagagggg tgaacttcac atccaacggc cctgtgatgc agaagaaaac actcggctgg
      421 gaggccttca ccgagacgct gtaccccgct gacggcggcc tggaaggcag aaacgacatg
      481 gccctgaagc tcgtgggcgg gagccatctg atcgcaaacg ccaagaccac atatagatcc
      541 aagaaacccg ctaagaacct caagatgcct ggcgtctact atgtggacta cagactggaa
      601 agaatcaagg aggccaacaa cgagacctac gtcgagcagc acgaggtggc agtggccaga
      661 tactgcgacc tccctagcaa actggggcac aagcttaatg gatccggcgc aacaaacttc
      721 tctctgctga aacaagccgg agatgtcgaa gagaatcctg gaccgatgga ctacaaggat
      781 gacgatgaca agggcgatta caaggatgac gacgataagg gagattacaa ggatgacgac
      841 gataaggctc gaggttatca aacaagtttg tacaaaaaag caggctccac catgttaagg
      901 aggattttac agaggactcc tgggagagtt ggctctcaag gttctgattt agattcatca
      961 gcaactccta taaacacagt ggacgtcaat aatgaaagct cttcagaggg tttcatatgt
     1021 ccccagtgta tgaaatctct tggatctgct gatgaacttt tcaaacatta tgaagctgtt
     1081 catgatgctg gtaatgactc aggtcatgga ggagagtcta atcttgcttt gaagcgagat
     1141 gatgtaacac tgctcagaca agaggtccaa gacctacagg cttcacttaa ggaagaaaaa
     1201 tggtactcgg aagaattaaa gaaggaatta gaaaaatatc aagggctgca gcagcaagag
     1261 gccaaacctg atgggttggt gactgattca tcagcagaac tacagtcttt ggaacagcaa
     1321 ttagaagaag cccaaacaga aaattttaat attaagcaaa tgaaagactt atttgaacag
     1381 aaagcagccc aacttgctac tgaaattgca gatataaagt caaagtatga tgaagaaagg
     1441 agtcttcgag aagctgctga acaaaaagtg acacgtctga cagaagaatt aaacaaagag
     1501 gcaactgtaa ttcaagatct gaagacggaa ctgcttcaga gacctggtat agaagatgtt
     1561 gccgtgctaa agaaagaact ggtccaagtt caaacactaa tggataacat gaccttggaa
     1621 cgtgagcgag aatctgaaaa actcaaagat gaatgcaaaa aattgcagtc acaatatgct
     1681 agctcagagg ccacaataag ccagctaagg agtgaacttg ccaaaggccc ccaggaagtt
     1741 gctgtatatg tacaggaact acaaaaactg aaaagttcag ttaatgaatt aacacaaaaa
     1801 aatcagacct tgacagaaaa cttgctgaaa aaagaacaag actatactaa gttagaggag
     1861 aaacataatg aagaatctgt gagtaaaaag aatattcagg caacccttca tcaaaaagac
     1921 ctagattgtc aacagcttca gtcaagattg tctgcatctg aaacctcact gcatagaata
     1981 catgtagaac taagtgaaaa aggagaagct actcaaaagc tcaaagaaga attatctgag
     2041 gtagagacca agtaccagca tctaaaggcg gagtttaagc agctacaaca acagagagaa
     2101 gaaaaggagc agcatgggtt acaactccaa agtgaaatta atcaattaca tagcaaactt
     2161 ctggagacag agcgccaact aggggaagct catggtaggc tgaaggaaca gagacagctt
     2221 tcaagtgaaa agttgatgga taaagaacaa caagtggctg atttacaact caaactttct
     2281 cggttagaag agcagttgaa ggaaaaagtt acaaattcta cagaattgca gcatcaatta
     2341 gataaaacaa agcaacagca tcaagaacaa caggctcttc agcaaagcac cacggcaaaa
     2401 cttcgagaag ctcagaatga tttggaacaa gttctacgtc aaattggcga taaggaccaa
     2461 aagatccaga accttgaagc tttattacag aagagtaaag aaaatatttc attactagaa
     2521 aaagaaagag aagatcttta tgcaaaaatt caggctggtg aaggagagac tgctgttctt
     2581 aaccagttac aagaaaaaaa ccatacacta caggagcaag taactcaact aacagagaag
     2641 ctgaagaatc agtcagaaag tcataaacaa gcccaggaga atttgcatga ccaggtacaa
     2701 gagcagaagg cacatcttag agctgcacaa gaccgtgtcc tttccctaga aactagtgtc
     2761 aatgaattaa atagtcaatt aaatgaaagc aaggagaagg tctcccagct tgacatacag
     2821 attaaagcca aaaccgaact attactatca gcagaagcag caaaaactgc tcaaagagct
     2881 gatcttcaga atcatttgga cacagctcaa aatgcattac aagataaaca gcaggagtta
     2941 aataagatta ctactcagtt ggatcaggtc actgcaaagt tacaagacaa gcaagaacat
     3001 tgcagtcagc tggaaagtca tcttaaagaa tataaagaga aatacctctc tttagaacag
     3061 aaaaccgaag agctagaagg tcaaattaag aaactagaag ctgatagtct tgaagttaaa
     3121 gcaagcaagg agcaggcttt gcaagatcta caacagcaaa gacagctgaa cacagattta
     3181 gagctcagag ccacagaatt gagtaaacaa cttgaaatgg agaaggaaat agtatccagt
     3241 acaagattgg atctacagaa aaaatctgaa gcccttgaaa gtatcaagca aaagcttacc
     3301 aagcaagagg aagaaaaaaa aatcctgaaa caagattttg aaactttaag tcaagaaaca
     3361 aagattcagc atgaggaatt gaataacaga attcaaacaa cagtaacaga actacaaaaa
     3421 gtgaaaatgg agaaagaagc tttaatgaca gagctttcta cagtaaagga caaactatca
     3481 aaagtttctg attctttgaa aaactctaaa agtgaatttg aaaaggagaa tcagaaagga
     3541 aaagccgcta tattagactt ggaaaaaact tgcaaagaat taaagcatca acttcaagtg
     3601 cagatggaaa acacacttaa ggaacagaag gaactgaaaa agtcacttga aaaagagaag
     3661 gaggcttctc atcagttgaa attggaactc aattcaatgc aggaacaact tatacaggcc
     3721 cagaatactt taaaacaaaa tgaaaaagaa gagcaacaac ttcaggggaa cataaatgag
     3781 ctaaagcaat caagtgaaca gaagaaaaaa caaattgaag cactccaagg agagcttaaa
     3841 attgctgttt tacagaagac agagcttgag aataaactac agcagcagtt aacacaggca
     3901 gcccaggaac ttgcagcaga gaaagagaaa atatcagtat tacaaaacaa ctatgaaaaa
     3961 agtcaggaaa ctttcaaaca gcttcaatct gatttctatg ggagggaatc tgaacttcta
     4021 gccaccaggc aagatcttaa gtctgtagaa gagaagcttt ctctagcaca ggaggacttg
     4081 atttcaaaca gaaatcaaat tggaaatcaa aataaattga ttcaagaact gaagactgcc
     4141 aaggctacat tggagcagga ttcagcaaag aaagaacagc aattgcagga gcgatgtaaa
     4201 gcactacaag acattcagaa agaaaagtca ctgaaagaaa aagaactggt aaatgagaag
     4261 tctaaattgg cagagataga agaaattaaa tgtagacaag aaaaagaaat cactaaacta
     4321 aacgaagaac tcaagtccca caaactagaa agcataaagg agataacaaa tcttaaagat
     4381 gctaaacagc ttctaattca gcagaaatta gaacttcaag gaaaagcgga ctccctgaag
     4441 gcagctgttg aacaggagaa gagaaatcag cagatactaa aagaccaggt gaaaaaggaa
     4501 gaagaggagc tgaagaaaga atttattgag aaagaagcta agttgcattc cgaaataaaa
     4561 gaaaaggaag taggaatgaa gaagcatgaa gaaaatgagg ctaaacttac catgcagatt
     4621 acagcattaa atgaaaactt aggcactgtg aagaaggagt ggcaatctag tcaacggaga
     4681 gttagtgagc ttgagaaaca aacggatgac ttacggggtg aaattgcagt attagaagca
     4741 acggttcaga ataatcaaga tgaaaggaga gcactactgg aaagatgtct taaaggagaa
     4801 ggtgaaatag aaaagcttca aaccaaagta ttagaattgc aaagaaagct ggataataca
     4861 actgcagcag tgcaggagct gggcagagaa aaccaatcac ttcagatcaa acatacacaa
     4921 gcgttgaata gaaagtgggc cgaagacaat gaagtacaaa actgtatggc ctgtgggaaa
     4981 ggcttttcag taacagtgag acggcatcac tgccgacagt gtggaaatat cttctgtgct
     5041 gaatgttcag ccaaaaatgc cttaactcct tcctccaaga agcctgttcg tgtctgtgat
     5101 gcatgtttca atgacttgca agga
//
