LOCUS       sequence-54242-e        2841 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 2841)
  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 2841)
  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-mRFP-T2A-FKBP-5-ptase
FEATURES             Location/Qualifiers
     source          1..2841
                     /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"
     CDS             430..1101
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKMR
                     LKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHST
                     GA"
     CDS             430..1101
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKMR
                     LKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHST
                     GA"
     primer_bind     complement(557..575)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     825..844
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1019..1042
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             1117..1170
                     /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      1177..1186
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1183..1185,1186..1506)
                     /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 gccaccctgg cctcctccga ggacgtcatc aaggagttca tgcgcttcaa ggtgcgcatg
      481 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
      541 gagggcaccc agaccgccaa gctgaaggtg accaagggcg gccccctgcc cttcgcctgg
      601 gacatcctgt cccctcagtt ccagtacggc tccaaggcct acgtgaagca ccccgccgac
      661 atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
      721 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc
      781 atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag
      841 aagaccatgg gctgggaggc ctccaccgag cggatgtacc ccgaggacgg cgccctgaag
      901 ggcgagatca agatgaggct gaagctgaag gacggcggcc actacgacgc cgaggtcaag
      961 accacctaca tggccaagaa gcccgtgcag ctgcccggcg cctacaagac cgacatcaag
     1021 ctggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga gcgcgccgag
     1081 ggccgccact ccaccggcgc cgtcgactcc gggagtgagg gcagaggaag tcttctaaca
     1141 tgcggtgacg tggaggagaa tcccggccca ccggtcgcca ccatgggagt gcaggtggaa
     1201 accatctccc caggagacgg gcgcaccttc cccaagcgcg gccagacctg cgtggtgcac
     1261 tacaccggga tgcttgaaga tggaaagaaa tttgattcct cccgggacag aaacaagccc
     1321 tttaagttta tgctaggcaa gcaggaggtg atccgaggct gggaagaagg ggttgcccag
     1381 atgagtgtgg gtcagagagc caaactgact atatctccag attatgccta tggtgccact
     1441 gggcacccag gcatcatccc accacatgcc actctcgtct tcgatgtgga gcttctaaaa
     1501 ctggaagctc gaggcgcagc agcgggagca ggaggagcag gtcgatcgga tcttgcagac
     1561 tacaagctcc gcgcgcagcc gctcctggtg cgggcccaca gcagcctggg ccccggccgg
     1621 ccgcggagcc ccctggcctg cgacgactgt tcccttcgct cggccaaatc ctccttcagc
     1681 ctcctggcgc ccatccgcag caaggacgtc cgcagcagga gttacctgga gggcagcctc
     1741 ctggccagcg gggccctgtt gggggcggat gagctggccc gctacttccc agaccggaac
     1801 gtggcactct tcgtggccac ctggaacatg cagggccaga aggagctccc gcccagcctg
     1861 gacgagttcc tgctcccagc cgaggccgac tatgcccagg acctgtatgt catcggggtc
     1921 caggagggct gttctgacag gcgggagtgg gagactcgtc tgcaggagac gctgggcccg
     1981 cactatgtgc tgctgtcctc ggcggcccac ggcgtgctct acatgtcgct cttcatccgc
     2041 agggacctca tctggttctg ctcagaggtg gagtgctcca cggtgaccac acgcatcgtg
     2101 tctcagatca agaccaaggg ggccttgggc atcagcttca ccttttttgg cacttccttc
     2161 ctcttcatca cgtcccactt cacctcaggt gacgggaagg tggcggagcg gctgctggac
     2221 tacaccagga ctgtacaagc cctggtcctg cccagaaatg tgcccgacac caacccctat
     2281 cgctccagcg cagcggacgt caccacccgc ttcgatgagg tgttctggtt tggagacttc
     2341 aacttccgcc tgagtggcgg gcgcacagtc gtggacgccc tcctgtgcca gggcctggtg
     2401 gtggacgtgc cggcgctgct gcagcacgac cagctcatcc gggagatgcg gaaagggtcc
     2461 atcttcaagg gcttccagga gccggacatc cacttcctcc catcatacaa gtttgacatc
     2521 gggaaggaca cgtacgacag cacctccaag cagaggacgc cctcatacac ggaccgcgtc
     2581 ttgtacagaa gccgccacaa gggtgacatc tgtcctgtga gctactcttc ctgccccggg
     2641 atcaagacgt ccgaccaccg ccctgtgtat ggcctcttcc gggtgaaagt gaggccgggg
     2701 cgagacaaca ttccgttggc agctggcaaa tttgatagag aactgtactt actaggaatt
     2761 aaaagacgga tttcgaagga gattcagagg cagcaagcac tacagagtca gaactccagc
     2821 accatcgcct ccgtttcttg a
//
