LOCUS       sequence-444504-         943 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Validates the expression and detection of epitope-tagged proteins of
            interest in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 943)
  AUTHORS   Robinson E, Alonso EB, A Waters J, Bileckyj C, D House C, A Johnston
            C, M Cripps R
  TITLE     pJoseph2: a family of plasmids as positive controls for bacterial 
            protein expression, transfections, and western blots.
  JOURNAL   Biotechniques. 2024;76(7):299-309. doi: 
            10.1080/07366205.2024.2343609. Epub 2024 May 12.
  PUBMED    39185782
REFERENCE   2  (bases 1 to 943)
  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: 
            "Biotechniques."; date: "2024"; volume: "76(7):299-309. doi"; pages:
            " 10.1080/07366205.2024.2343609. Epub 2024 May 12"
COMMENT     Sequence Label: pMH-HA/Joseph2-Multitagged Plasmid
FEATURES             Location/Qualifiers
     source          1..943
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     gap             2..11
                     /estimated_length=10
     regulatory      77..86
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             104..130
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             131..157
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             176..196
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    197..221
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             236..325
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /label=3xMyc
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
     primer_bind     complement(386..414)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
ORIGIN
        1 gnnnnnnnnn nagcagagct ttctggctaa ctagagaacc cactgcttac tggcacgtgg
       61 aaattaatac gacgtggcca ccatggccca ggtgcagctg cagtacccat acgacgtgcc
      121 agactacgca tacccatacg acgtgccaga ctacgcactg caggtcgacc tcgaggaaaa
      181 cctgtatttt cagggcacaa gtttgtacaa aaaagcaggc ttcggcagcg gaagcgagca
      241 aaagctcatc tcggaggaag atctagaaca gaagctaata tcagaggagg acctagagca
      301 aaaacttatt tcggaagaag atctgggcag cggatccggt tcgggaagcg gtagtatgag
      361 taaaggagaa gaacttttca ctggagttgt cccaattctt gttgaattag atggcgatgt
      421 taatgggcaa aaattctctg tcagtggaga gggtgaaggt gatgcaacat acggaaaact
      481 tacccttaaa tttatttgca ctactgggaa gctacctgtt ccatggccaa cacttgtcac
      541 tactttgact tatggtgttc aatgcttttc aagataccca gatcatatga aacagcatga
      601 ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa agaactatat tttacaaaga
      661 tgacgggaac tacaagacac gtgctgaagt caagtttgaa ggtgataccc ttgttaatag
      721 aatcgagtta aaaggtattg attttaaaga agatggaaac attcttggac acaaaatgga
      781 atacaactat aactcacata atgtatacat catggcagac aaaccaaaga atggaatcaa
      841 agttaacttc aaaattagac acaacattaa agatggaagc gttcaattag cagaccatta
      901 tcaacaaaat actccaattg gcgatggccc tgtcctttta cca
//
