LOCUS       sequence-419289-        1850 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Endogenously tag mouse 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 1850)
  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 1850)
  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: pTwist Amp msTMEM115-TEV-3C-P2A-mScarlet-I HDR...
FEATURES             Location/Qualifiers
     source          1..1850
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             518..538
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             539..619
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             629..685
                     /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             686..1381
                     /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(819..837)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1087..1106
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 actgaaacac gccactcctg acagaagcac tacacccacc cgtctttagc tagcaaactc
       61 agaagtatcc gtggtggctg gcctgctagg taggctgcca acatctaggt agccttctca
      121 gtctggaggt gccttgggtc ctcattcaat tgccaaagca gatcctggcc caggctctta
      181 gacgcggcac ggggatggga aacgtgggat ttggaggcaa acctctttgg taggaggatt
      241 tggccttatt gaagaaggtt gcagagtatg ttggtctccc aaacccagtg taacctgcct
      301 tttctcttct ggctccaggc aactagccct aaaggctctc aatgagcggc tgaagagagt
      361 ggaggatcag tcagcctggc ccagcatgga tgatgacgaa gaggaagctg gggcaaaaac
      421 ggacagtcct ctgcccttag aagaagcttc cacgccaccc ggaaaagtga cagtgccgga
      481 atccagtctc atcaccttgg agacagctcc cctactcgaa aacctgtact tccagggcta
      541 cccatacgat gttccagatt acgcttatcc ttatgacgta cctgactatg cataccctta
      601 tgatgtacca gactacgctg gatccggcgc aacaaacttc tctctgctga aacaagccgg
      661 agatgtcgaa gagaatcctg gaccgatggt gagcaagggc gaggcagtga tcaaggagtt
      721 catgcggttc aaggtgcaca tggagggctc catgaacggc cacgagttcg agatcgaggg
      781 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
      841 tggccccctg cccttctcct gggacatcct gtcccctcag ttcatgtacg gctccagggc
      901 cttcatcaag caccccgccg acatccccga ctactataag cagtccttcc ccgagggctt
      961 caagtgggag cgcgtgatga acttcgagga cggcggcgcc gtgaccgtga cccaggacac
     1021 ctccctggag gacggcaccc tgatctacaa ggtgaagctc cgcggcacca acttccctcc
     1081 tgacggcccc gtaatgcaga agaagacaat gggctgggaa gcgtccaccg agcggttgta
     1141 ccccgaggac ggcgtgctga agggcgacat taagatggcc ctgcgcctga aggacggcgg
     1201 ccgctacctg gcggacttca agaccaccta caaggccaag aagcccgtgc agatgcccgg
     1261 cgcctacaac gtcgaccgca agttggacat cacctcccac aacgaggact acaccgtggt
     1321 ggaacagtac gaacgctccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     1381 gtagaccact ttgagtgcag ttcttctact cccatgcctt cccgatccgt ctcggctact
     1441 gcagagcact gactgctctg aggagaggga agagggtctg ctgggggttc ctgtggcctt
     1501 ccgctgtgtc tggacaacac taacacagga cccttgttgc tacctggttc tgactccaga
     1561 caaccacaat gccaggtacg gggtctctga gcagcagcca gtgcagatcc ccatttgcag
     1621 taagattgta cctcagcggt acaattcctc accaaggaag tggctgcagg gacactggtg
     1681 ccacggctcc tgggccagcc cttatgtctg aaactggttg ccaacagccc cgagccaagg
     1741 caaggatttg tgtttgccaa aaatgttctg gggacccagc cagtgtgggt ctgaacatcc
     1801 ccgcccaccc ccagactgca ttctcatcag ggtttcttgt gcccttctgc
//
