LOCUS       sequence-54244-e        2883 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2883)
  AUTHORS   Toth DJ, Toth JT, Gulyas G, Balla A, Balla T, Hunyady L, Varnai P
  TITLE     Acute depletion of plasma membrane phosphatidylinositol 
            4,5-bisphosphate impairs specific steps in endocytosis of the 
            G-protein-coupled receptor.
  JOURNAL   J Cell Sci. 2012 May 1;125(Pt 9):2185-97. Epub 2012 Feb 22.
  PUBMED    22357943
REFERENCE   2  (bases 1 to 2883)
  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: "J Cell 
            Sci."; date: "2012-05-1"; volume: "125(Pt 9)"; pages: "2185-97. Epub
            2012 Feb 22"
COMMENT     Sequence Label: PM-FRB-Cerulean-T2A-FKBP-5-ptase
FEATURES             Location/Qualifiers
     source          1..2883
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(71..88)
                     /codon_start=1
                     /product="HQHQHQ metal affinity tag"
                     /label=HQ tag
                     /note="The HQ tag binds to nickel and other metal ions much
                     like the 6xHis tag, but may allow elution at lower 
                     concentrations of imidazole (Godat et al., 2008)."
                     /translation="HQHQHQ"
     CDS             103..381
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISK"
     CDS             388..414
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      421..430
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(427..429,430..432,433..1143)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(472..493)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(733..752)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1080..1101
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1159..1212
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     regulatory      1219..1228
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1225..1227,1228..1548)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
ORIGIN
        1 atgggatgta taaaatcaaa agggaaagac agcgcgggag cagatcccac cagatctgcg
       61 aattcaggtg ctggtgctgg tgctggtgct atcctatcta gaatcctctg gcatgagatg
      121 tggcatgaag gcctggaaga ggcatctcgt ttgtactttg gggaaaggaa cgtgaaaggc
      181 atgtttgagg tgctggagcc cttgcatgct atgatggaac ggggccccca gactctgaag
      241 gaaacatcct ttaatcaggc ctatggtcga gatttaatgg aggcccaaga gtggtgcagg
      301 aagtacatga aatcagggaa tgtcaaggac ctcacccaag cctgggacct ctattatcat
      361 gtgttccgac gaatctcaaa gactagttat ccgtacgacg taccagacta cgcaccggtc
      421 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
      481 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
      541 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
      601 cccaccctcg tgaccaccct gacctggggc gtgcagtgct tcgcccgcta ccccgaccac
      661 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
      721 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
      781 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
      841 gggcacaagc tggagtacaa cgccatcagc gacaacgtct atatcaccgc cgacaagcag
      901 aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
      961 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     1021 aaccactacc tgagcaccca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac
     1081 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     1141 aaggtcgact ccgggagtga gggcagagga agtcttctaa catgcggtga cgtggaggag
     1201 aatcccggcc caccggtcgc caccatggga gtgcaggtgg aaaccatctc cccaggagac
     1261 gggcgcacct tccccaagcg cggccagacc tgcgtggtgc actacaccgg gatgcttgaa
     1321 gatggaaaga aatttgattc ctcccgggac agaaacaagc cctttaagtt tatgctaggc
     1381 aagcaggagg tgatccgagg ctgggaagaa ggggttgccc agatgagtgt gggtcagaga
     1441 gccaaactga ctatatctcc agattatgcc tatggtgcca ctgggcaccc aggcatcatc
     1501 ccaccacatg ccactctcgt cttcgatgtg gagcttctaa aactggaagc tcgaggcgca
     1561 gcagcgggag caggaggagc aggtcgatcg gatcttgcag actacaagct ccgcgcgcag
     1621 ccgctcctgg tgcgggccca cagcagcctg ggccccggcc ggccgcggag ccccctggcc
     1681 tgcgacgact gttcccttcg ctcggccaaa tcctccttca gcctcctggc gcccatccgc
     1741 agcaaggacg tccgcagcag gagttacctg gagggcagcc tcctggccag cggggccctg
     1801 ttgggggcgg atgagctggc ccgctacttc ccagaccgga acgtggcact cttcgtggcc
     1861 acctggaaca tgcagggcca gaaggagctc ccgcccagcc tggacgagtt cctgctccca
     1921 gccgaggccg actatgccca ggacctgtat gtcatcgggg tccaggaggg ctgttctgac
     1981 aggcgggagt gggagactcg tctgcaggag acgctgggcc cgcactatgt gctgctgtcc
     2041 tcggcggccc acggcgtgct ctacatgtcg ctcttcatcc gcagggacct catctggttc
     2101 tgctcagagg tggagtgctc cacggtgacc acacgcatcg tgtctcagat caagaccaag
     2161 ggggccttgg gcatcagctt cacctttttt ggcacttcct tcctcttcat cacgtcccac
     2221 ttcacctcag gtgacgggaa ggtggcggag cggctgctgg actacaccag gactgtacaa
     2281 gccctggtcc tgcccagaaa tgtgcccgac accaacccct atcgctccag cgcagcggac
     2341 gtcaccaccc gcttcgatga ggtgttctgg tttggagact tcaacttccg cctgagtggc
     2401 gggcgcacag tcgtggacgc cctcctgtgc cagggcctgg tggtggacgt gccggcgctg
     2461 ctgcagcacg accagctcat ccgggagatg cggaaagggt ccatcttcaa gggcttccag
     2521 gagccggaca tccacttcct cccatcatac aagtttgaca tcgggaagga cacgtacgac
     2581 agcacctcca agcagaggac gccctcatac acggaccgcg tcttgtacag aagccgccac
     2641 aagggtgaca tctgtcctgt gagctactct tcctgccccg ggatcaagac gtccgaccac
     2701 cgccctgtgt atggcctctt ccgggtgaaa gtgaggccgg ggcgagacaa cattccgttg
     2761 gcagctggca aatttgatag agaactgtac ttactaggaa ttaaaagacg gatttcgaag
     2821 gagattcaga ggcagcaagc actacagagt cagaactcca gcaccatcgc ctccgtttct
     2881 tga
//
