LOCUS       sequence-419285-        1950 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Stably express the GolgiTag (GolgiIP handle) in mammalian cells 
            (human ORF). eGFP for cell sorting desired expression levels..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1950)
  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 1950)
  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 hTMEM115-3C-3xHA-P2A-eGFP
FEATURES             Location/Qualifiers
     source          1..1950
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1054..1077
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1078..1104
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1108..1134
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1138..1164
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1174..1230
                     /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             join(1231..1233,1234..1236,1237..1950)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1231..1233,1234..1236,1237..1947)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1276..1297)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1537..1556)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1884..1905
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 atgcaacgtg ccctgccagg cgcccgccag cacttggggg ccattctggc cagcgccagc
       61 gtggtggtga aggctctgtg tgcggcggta ctattcctct acctgctctc cttcgccgtg
      121 gacacaggct gcctggcggt caccccgggc tacctctttc ctcccaactt ctggatctgg
      181 accctggcca cccatgggct gatggagcag catgtgtggg acgtggccat cagcctgaca
      241 acggtggtgg tggccgggcg tttgctggag cccctctggg gggccttgga gctgctcatc
      301 ttcttctcag tggtgaatgt gtctgtaggg ctgctggggg ccttcgccta cctcctcacc
      361 tacatggctt ccttcaacct ggtctacctg ttcactgtcc gtatccacgg cgccttgggc
      421 ttcctaggtg gcgtcctggt ggcactcaag caaaccatgg gggactgtgt ggtcctgcga
      481 gtgccccagg tgcgcgtcag tgtgatgccc atgctgctgc tggcgctgct gctcctgctg
      541 cggctcgcca cgctgctcca gagcccggcg ctggcttcct atggcttcgg gctgctctcc
      601 agttgggtat atcttcgctt ctaccagcgc catagccggg gccgagggga catggctgac
      661 cactttgctt tcgccacttt cttccctgag atcctgcagc ctgtggtggg tttgctggcg
      721 aacttggtgc acagcctcct ggtgaaggta aagatatgcc agaagacggt gaagcgctac
      781 gatgtgggtg ccccatcctc catcaccatc agcctgccag gcacagaccc tcaagacgcc
      841 gagcggagaa ggcaactggc cctgaaggca ctcaatgagc ggctgaagag agtggaagac
      901 cagtccatct ggcccagcat ggatgatgat gaagaggagt ctggggccaa ggtggacagc
      961 cccctgccct cagacaaagc tcccacaccc ccagggaagg gggctgcccc agaatccagt
     1021 ctaatcacct tcgaggcagc tcccccgacg ctgcttgagg tgctctttca gggaccctac
     1081 ccgtatgatg ttccggatta cgctggctat ccctacgacg tgcccgacta tgccgggtac
     1141 ccctatgacg tcccagacta cgcaggatcc ggcgcaacaa acttctctct gctgaaacaa
     1201 gccggagatg tcgaagagaa tcctggaccg atggtgagca agggcgagga gctgttcacc
     1261 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     1321 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     1381 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     1441 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     1501 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     1561 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     1621 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     1681 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     1741 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     1801 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     1861 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     1921 actctcggca tggacgagct gtacaagtaa
//
