LOCUS       sequence-138454-         902 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses HA and YFP tagged PRAS40.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 902)
  AUTHORS   Jain A, Arauz E, Aggarwal V, Ikon N, Chen J, Ha T
  TITLE     Stoichiometry and assembly of mTOR complexes revealed by 
            single-molecule pulldown.
  JOURNAL   Proc Natl Acad Sci U S A. 2014 Dec 16;111(50):17833-8. doi: 
            10.1073/pnas.1419425111. Epub 2014 Dec 1.
  PUBMED    25453101
REFERENCE   2  (bases 1 to 902)
  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"; doi: 
            "10.1073/pnas.1419425111"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2014-12-16- 16"; volume: "111"; issue: "50"; pages: "17833-8"
COMMENT     Sequence Label: pRK5-HA-YFP-PRAS40
FEATURES             Location/Qualifiers
     source          1..902
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      84..93
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             96..122
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      153..162
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(159..161,162..164,165..875)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(204..225)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(465..484)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     812..833
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 tccactccca ggtccaactg cacctcggtt ctatcgattg aattcccgaa ccgacagtcg
       61 gtctcttcac caaggccatt cgcgccacca tgggctaccc ctatgatgtg cctgactacg
      121 cagatcgcaa tggtggtggt ggtggaccgg tcgccaccat ggtgagcaag ggcgaggagc
      181 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
      241 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
      301 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ttcggctacg
      361 gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
      421 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
      481 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
      541 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
      601 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
      661 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
      721 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccgccc
      781 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
      841 ccgggatcac tctcggcatg gacgaactgt acaagtccgg actcagatct ggtggaaggc
      901 ct
//
