LOCUS       sequence-419287-        1973 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Endogenously tag human TMEM115 with a C-terminal 3xHA tag in 
            addition to cytosolic mScarlet-I for cell sorting..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1973)
  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 1973)
  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: hTMEM115-TEV-3xHA-P2A-mScarlet HDR template
FEATURES             Location/Qualifiers
     source          1..1973
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             646..666
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             667..693
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             697..723
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             727..753
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             763..819
                     /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             820..1515
                     /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(953..971)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1221..1240
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 caggagaggg ggagaattaa taacccttcc cttggatgtg atgccagcac ccaggcagaa
       61 actcctggaa actccagaac ttgccgttcc acccatctgc cctagggcac tagttccact
      121 gacccttaga taccaccctc agaaccattc ctggtggctg gcctcgtggg cagtctggca
      181 gacacccagc tgaccttccc atcccggagg tgcctcagct atggagggaa gattcttaga
      241 gcctttgggt tatcattctg atggcagagt gggtcctaaa ccaatctctg gggagtccca
      301 ggctctgcat ggtgaagggg aaacatagga ttctggggga ggagtctagg tggtacggtg
      361 aggagtcagc ctgctgcgga agggtagagg gcatgagcca aggccaaacc cagagtgacc
      421 tttcctctct cctctggccc caggcaactg gcactcaaag ccctcaatga gcggctgaag
      481 agagtggaag accagtccat ctggcccagc atggatgatg atgaagagga gtctggggcc
      541 aaggtggaca gccccctgcc ctcagacaaa gctcccacac ccccagggaa gggggctgcc
      601 ccagaatcca gtctaatcac cttcgaggca gctcccccga cgctggaaaa cctgtacttc
      661 cagggctacc cgtatgatgt tccggattac gctggctatc cctacgacgt gcccgactat
      721 gccgggtacc cctatgacgt cccagactac gcaggatccg gcgcaacaaa cttctctctg
      781 ctgaaacaag ccggagatgt cgaagagaat cctggaccga tggtgagcaa gggcgaggca
      841 gtgatcaagg agttcatgcg gttcaaggtg cacatggagg gctccatgaa cggccacgag
      901 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
      961 aaggtgacca agggtggccc cctgcccttc tcctgggaca tcctgtcccc tcagttcatg
     1021 tacggctcca gggccttcat caagcacccc gccgacatcc ccgactacta taagcagtcc
     1081 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgccgtgacc
     1141 gtgacccagg acacctccct ggaggacggc accctgatct acaaggtgaa gctccgcggc
     1201 accaacttcc ctcctgacgg ccccgtaatg cagaagaaga caatgggctg ggaagcgtcc
     1261 accgagcggt tgtaccccga ggacggcgtg ctgaagggcg acattaagat ggccctgcgc
     1321 ctgaaggacg gcggccgcta cctggcggac ttcaagacca cctacaaggc caagaagccc
     1381 gtgcagatgc ccggcgccta caacgtcgac cgcaagttgg acatcacctc ccacaacgag
     1441 gactacaccg tggtggaaca gtacgaacgc tccgagggcc gccactccac cggcggcatg
     1501 gacgagctgt acaagtaact ccagaccacc ttgagtgtgg cacctcccct cccaagcccc
     1561 ccttgacatc ctctcagcta ctccagggca cctgactgct ctgaggagag ggaagaaggc
     1621 ctgctggggc tttccatggc cttctgctgt ttctcgccaa cactacccag gactcttgct
     1681 acctggttcc aactccagac aaccactatg ccaggcccgg agcctctgag gcatcggcca
     1741 gtccaggccc tcatctgagg taagaatgta catcagctgg cagccccaag caagtggctg
     1801 cagggacact gatgccacag ctcctgggcc ggccctcaca tctgaaactg gttgccgaga
     1861 gccctgagcc aaggcaagga tttgccaaaa atgttctggg ggcccagcaa atgcaggagc
     1921 cgacctgggg ctgcacatcc ctgcccatcc ccagaaagac tgttcctgtc agg
//
