LOCUS       sequence-419283-        5121 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Express the human EndoTag (3xFLAG-EEA1) for EndoIPs. mScarlet-I to 
            sort for even expression levels..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5121)
  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 5121)
  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 mScarlet-I-P2A-3xFLAG-hEEA1
FEATURES             Location/Qualifiers
     source          1..5121
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1..696
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(134..152)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     402..421
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             706..762
                     /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             766..789
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             793..816
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             820..843
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    859..883
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             889..5121
                     /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 atggtgagca agggcgaggc agtgatcaag gagttcatgc ggttcaaggt gcacatggag
       61 ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
      121 ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt ctcctgggac
      181 atcctgtccc ctcagttcat gtacggctcc agggccttca tcaagcaccc cgccgacatc
      241 cccgactact ataagcagtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
      301 gaggacggcg gcgccgtgac cgtgacccag gacacctccc tggaggacgg caccctgatc
      361 tacaaggtga agctccgcgg caccaacttc cctcctgacg gccccgtaat gcagaagaag
      421 acaatgggct gggaagcgtc caccgagcgg ttgtaccccg aggacggcgt gctgaagggc
      481 gacattaaga tggccctgcg cctgaaggac ggcggccgct acctggcgga cttcaagacc
      541 acctacaagg ccaagaagcc cgtgcagatg cccggcgcct acaacgtcga ccgcaagttg
      601 gacatcacct cccacaacga ggactacacc gtggtggaac agtacgaacg ctccgagggc
      661 cgccactcca ccggcggcat ggacgagctg tacaagggat ccggcgcaac aaacttctct
      721 ctgctgaaac aagccggaga tgtcgaagag aatcctggac cgatggacta caaggatgac
      781 gatgacaagg gcgattacaa ggatgacgac gataagggag attacaagga tgacgacgat
      841 aaggctcgag gttatcaaac aagtttgtac aaaaaagcag gctccaccat gttaaggagg
      901 attttacaga ggactcctgg gagagttggc tctcaaggtt ctgatttaga ttcatcagca
      961 actcctataa acacagtgga cgtcaataat gaaagctctt cagagggttt catatgtccc
     1021 cagtgtatga aatctcttgg atctgctgat gaacttttca aacattatga agctgttcat
     1081 gatgctggta atgactcagg tcatggagga gagtctaatc ttgctttgaa gcgagatgat
     1141 gtaacactgc tcagacaaga ggtccaagac ctacaggctt cacttaagga agaaaaatgg
     1201 tactcggaag aattaaagaa ggaattagaa aaatatcaag ggctgcagca gcaagaggcc
     1261 aaacctgatg ggttggtgac tgattcatca gcagaactac agtctttgga acagcaatta
     1321 gaagaagccc aaacagaaaa ttttaatatt aagcaaatga aagacttatt tgaacagaaa
     1381 gcagcccaac ttgctactga aattgcagat ataaagtcaa agtatgatga agaaaggagt
     1441 cttcgagaag ctgctgaaca aaaagtgaca cgtctgacag aagaattaaa caaagaggca
     1501 actgtaattc aagatctgaa gacggaactg cttcagagac ctggtataga agatgttgcc
     1561 gtgctaaaga aagaactggt ccaagttcaa acactaatgg ataacatgac cttggaacgt
     1621 gagcgagaat ctgaaaaact caaagatgaa tgcaaaaaat tgcagtcaca atatgctagc
     1681 tcagaggcca caataagcca gctaaggagt gaacttgcca aaggccccca ggaagttgct
     1741 gtatatgtac aggaactaca aaaactgaaa agttcagtta atgaattaac acaaaaaaat
     1801 cagaccttga cagaaaactt gctgaaaaaa gaacaagact atactaagtt agaggagaaa
     1861 cataatgaag aatctgtgag taaaaagaat attcaggcaa cccttcatca aaaagaccta
     1921 gattgtcaac agcttcagtc aagattgtct gcatctgaaa cctcactgca tagaatacat
     1981 gtagaactaa gtgaaaaagg agaagctact caaaagctca aagaagaatt atctgaggta
     2041 gagaccaagt accagcatct aaaggcggag tttaagcagc tacaacaaca gagagaagaa
     2101 aaggagcagc atgggttaca actccaaagt gaaattaatc aattacatag caaacttctg
     2161 gagacagagc gccaactagg ggaagctcat ggtaggctga aggaacagag acagctttca
     2221 agtgaaaagt tgatggataa agaacaacaa gtggctgatt tacaactcaa actttctcgg
     2281 ttagaagagc agttgaagga aaaagttaca aattctacag aattgcagca tcaattagat
     2341 aaaacaaagc aacagcatca agaacaacag gctcttcagc aaagcaccac ggcaaaactt
     2401 cgagaagctc agaatgattt ggaacaagtt ctacgtcaaa ttggcgataa ggaccaaaag
     2461 atccagaacc ttgaagcttt attacagaag agtaaagaaa atatttcatt actagaaaaa
     2521 gaaagagaag atctttatgc aaaaattcag gctggtgaag gagagactgc tgttcttaac
     2581 cagttacaag aaaaaaacca tacactacag gagcaagtaa ctcaactaac agagaagctg
     2641 aagaatcagt cagaaagtca taaacaagcc caggagaatt tgcatgacca ggtacaagag
     2701 cagaaggcac atcttagagc tgcacaagac cgtgtccttt ccctagaaac tagtgtcaat
     2761 gaattaaata gtcaattaaa tgaaagcaag gagaaggtct cccagcttga catacagatt
     2821 aaagccaaaa ccgaactatt actatcagca gaagcagcaa aaactgctca aagagctgat
     2881 cttcagaatc atttggacac agctcaaaat gcattacaag ataaacagca ggagttaaat
     2941 aagattacta ctcagttgga tcaggtcact gcaaagttac aagacaagca agaacattgc
     3001 agtcagctgg aaagtcatct taaagaatat aaagagaaat acctctcttt agaacagaaa
     3061 accgaagagc tagaaggtca aattaagaaa ctagaagctg atagtcttga agttaaagca
     3121 agcaaggagc aggctttgca agatctacaa cagcaaagac agctgaacac agatttagag
     3181 ctcagagcca cagaattgag taaacaactt gaaatggaga aggaaatagt atccagtaca
     3241 agattggatc tacagaaaaa atctgaagcc cttgaaagta tcaagcaaaa gcttaccaag
     3301 caagaggaag aaaaaaaaat cctgaaacaa gattttgaaa ctttaagtca agaaacaaag
     3361 attcagcatg aggaattgaa taacagaatt caaacaacag taacagaact acaaaaagtg
     3421 aaaatggaga aagaagcttt aatgacagag ctttctacag taaaggacaa actatcaaaa
     3481 gtttctgatt ctttgaaaaa ctctaaaagt gaatttgaaa aggagaatca gaaaggaaaa
     3541 gccgctatat tagacttgga aaaaacttgc aaagaattaa agcatcaact tcaagtgcag
     3601 atggaaaaca cacttaagga acagaaggaa ctgaaaaagt cacttgaaaa agagaaggag
     3661 gcttctcatc agttgaaatt ggaactcaat tcaatgcagg aacaacttat acaggcccag
     3721 aatactttaa aacaaaatga aaaagaagag caacaacttc aggggaacat aaatgagcta
     3781 aagcaatcaa gtgaacagaa gaaaaaacaa attgaagcac tccaaggaga gcttaaaatt
     3841 gctgttttac agaagacaga gcttgagaat aaactacagc agcagttaac acaggcagcc
     3901 caggaacttg cagcagagaa agagaaaata tcagtattac aaaacaacta tgaaaaaagt
     3961 caggaaactt tcaaacagct tcaatctgat ttctatggga gggaatctga acttctagcc
     4021 accaggcaag atcttaagtc tgtagaagag aagctttctc tagcacagga ggacttgatt
     4081 tcaaacagaa atcaaattgg aaatcaaaat aaattgattc aagaactgaa gactgccaag
     4141 gctacattgg agcaggattc agcaaagaaa gaacagcaat tgcaggagcg atgtaaagca
     4201 ctacaagaca ttcagaaaga aaagtcactg aaagaaaaag aactggtaaa tgagaagtct
     4261 aaattggcag agatagaaga aattaaatgt agacaagaaa aagaaatcac taaactaaac
     4321 gaagaactca agtcccacaa actagaaagc ataaaggaga taacaaatct taaagatgct
     4381 aaacagcttc taattcagca gaaattagaa cttcaaggaa aagcggactc cctgaaggca
     4441 gctgttgaac aggagaagag aaatcagcag atactaaaag accaggtgaa aaaggaagaa
     4501 gaggagctga agaaagaatt tattgagaaa gaagctaagt tgcattccga aataaaagaa
     4561 aaggaagtag gaatgaagaa gcatgaagaa aatgaggcta aacttaccat gcagattaca
     4621 gcattaaatg aaaacttagg cactgtgaag aaggagtggc aatctagtca acggagagtt
     4681 agtgagcttg agaaacaaac ggatgactta cggggtgaaa ttgcagtatt agaagcaacg
     4741 gttcagaata atcaagatga aaggagagca ctactggaaa gatgtcttaa aggagaaggt
     4801 gaaatagaaa agcttcaaac caaagtatta gaattgcaaa gaaagctgga taatacaact
     4861 gcagcagtgc aggagctggg cagagaaaac caatcacttc agatcaaaca tacacaagcg
     4921 ttgaatagaa agtgggccga agacaatgaa gtacaaaact gtatggcctg tgggaaaggc
     4981 ttttcagtaa cagtgagacg gcatcactgc cgacagtgtg gaaatatctt ctgtgctgaa
     5041 tgttcagcca aaaatgcctt aactccttcc tccaagaagc ctgttcgtgt ctgtgatgca
     5101 tgtttcaatg acttgcaagg a
//
