LOCUS       sequence-419286-        1947 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Stably express the GolgiTag (GolgiIP handle) in mammalian cells 
            (mouse ORF). eGFP for cell sorting desired expression levels..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1947)
  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 1947)
  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 msTMEM115-3C-3xHA-P2A-eGFP
FEATURES             Location/Qualifiers
     source          1..1947
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1051..1074
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1075..1101
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1105..1131
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1135..1161
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1171..1227
                     /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(1228..1230,1231..1233,1234..1947)
                     /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(1228..1230,1231..1233,1234..1944)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1273..1294)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1534..1553)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1881..1902
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 atgcaaaggg ccttgccagg agccagacaa cacctaggtg ctatcttggc atccgcgtcc
       61 gtcgttgtta aggcactctg tgctgttgta ctcttccttt atctcctttc cttcgctgtt
      121 gacactggtt gtctcgccgt cactcctggt tacctcttcc cacccaattt ctggatctgg
      181 acactcgcta ctcacggact aatggagcaa cacgtttggg acgtcgccat ctccttggca
      241 actgtcgttg ttgcaggtag gttgctagag cccctttggg gagctttgga gcttctaata
      301 ttcttctcag tcgtcaatgt ttctgtagga ttgctcgggg ccctcgcata tctcctcaca
      361 tacatggcat ctttcaatct tgtttacctc ttcactatcc gaatccacgg agccctcggg
      421 ttccttggcg gtgtactcgt agcattgaag caaacaatgg gtgactgtgt tgtccttcgg
      481 gttcctcaag tccgagtctc agtagtcccc atgctcttgc tagcactatt gctccttttg
      541 agacttgcca ctttgctaca atctcctgct cttgcctcat acggtttcgg actcctctcc
      601 tcttgggttt acttgaggtt ctaccaaagg cactctagag ggagaggaga catggccgac
      661 cacttcgctt tcgccacatt cttccccgag atccttcaac ccgtcgttgg acttttggcc
      721 aatttggttc acggactact tgttaaggta aagatctgtc aaaagactgt aaagaggtac
      781 gacgttgggg ccccaagttc catcacaatc agtttgccag ggacagaccc acaagacgca
      841 gagcgtagga gacaacttgc tcttaaggca ttgaatgaga ggcttaagcg ggtagaggac
      901 caatctgctt ggccctctat ggacgacgac gaggaagagg ccggggcaaa gactgactct
      961 ccactccctc tcgaggaagc ctcaactcca cccggaaagg ttacagtacc cgagtccagc
     1021 ttgatcactt tggagacggc tcccttgttg cttgaggtgc tctttcaggg accctacccg
     1081 tatgatgttc cggattacgc tggctatccc tacgacgtgc ccgactatgc cgggtacccc
     1141 tatgacgtcc cagactacgc aggatccggc gcaacaaact tctctctgct gaaacaagcc
     1201 ggagatgtcg aagagaatcc tggaccgatg gtgagcaagg gcgaggagct gttcaccggg
     1261 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     1321 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     1381 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
     1441 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     1501 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     1561 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     1621 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     1681 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     1741 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     1801 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
     1861 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     1921 ctcggcatgg acgagctgta caagtaa
//
