LOCUS       sequence-125337-        1114 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses APEX2 in the cytosol of mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1114)
  AUTHORS   Lam SS, Martell JD, Kamer KJ, Deerinck TJ, Ellisman MH, Mootha VK, 
            Ting AY
  TITLE     Directed evolution of APEX2 for electron microscopy and proximity 
            labeling.
  JOURNAL   Nat Methods. 2014 Nov 24. doi: 10.1038/nmeth.3179.
  PUBMED    25419960
REFERENCE   2  (bases 1 to 1114)
  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: "Nat 
            Methods. 2014 Nov 24. doi: 10.1038/nmeth.3179."
COMMENT     Sequence Label: pcDNA3 APEX2-NES
FEATURES             Location/Qualifiers
     source          1..1114
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     61..80
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        61..79
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      99..108
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             108..131
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             132..878
                     /codon_start=1
                     /product="soybean ascorbate peroxidase, engineered to serve
                     as a monomeric tag for electron microscopy and live-cell 
                     proteomics (Lam et al., 2015)"
                     /label=APEX2
                     /note="more active than the original APEX due to the A134P 
                     mutation"
                     /translation="GKSYPTVSADYQDAVEKAKKKLRGFIAEKRCAPLMLRLAFHSAGT
                     FDKGTKTGGPFGTIKHPAELAHSANNGLDIAVRLLEPLKAEFPILSYADFYQLAGVVAV
                     EVTGGPKVPFHPGREDKPEPPPEGRLPDPTKGSDHLRDVFGKAMGLTDQDIVALSGGHT
                     IGAAHKERSGFEGPWTSNPLIFDNSYFTELLSGEKEGLLQLPSDKALLSDPVFRPLVDK
                     YAADEDAFFADYAEAHQKLSELGFADA"
     CDS             885..911
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     promoter        complement(948..966)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(949..966)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(986..1003)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    992..1103
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 ctatataagc agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat
       61 taatacgact cactataggg agacccaagc ttgcggccgc caccatggac tacaaggatg
      121 acgacgataa gggaaagtct tacccaactg tgagtgctga ttaccaggac gccgttgaga
      181 aggcgaagaa gaagctcaga ggcttcatcg ctgagaagag atgcgctcct ctaatgctcc
      241 gtttggcatt ccactctgct ggaacctttg acaagggcac gaagaccggt ggacccttcg
      301 gaaccatcaa gcaccctgcc gaactggctc acagcgctaa caacggtctt gacatcgctg
      361 ttaggctttt ggagccactc aaggcggagt tccctatttt gagctacgcc gatttctacc
      421 agttggctgg cgttgttgcc gttgaggtca cgggtggacc taaggttcca ttccaccctg
      481 gaagagagga caagcctgag ccaccaccag agggtcgctt gcccgatccc actaagggtt
      541 ctgaccattt gagagatgtg tttggcaaag ctatggggct tactgaccaa gatatcgttg
      601 ctctatctgg gggtcacact attggagctg cacacaagga gcgttctgga tttgagggtc
      661 cctggacctc taatcctctt attttcgaca actcatactt cacggagttg ttgagtggtg
      721 agaaggaagg tctccttcag ctaccttctg acaaggctct tttgtctgac cctgtattcc
      781 gccctctcgt tgacaaatat gcagcggacg aagatgcctt ctttgctgat tacgctgagg
      841 ctcaccaaaa gctttccgag cttgggtttg ctgatgccct gcagctgcct cccctggagc
      901 gcctgaccct ggactaatag ctcgagcatg catctagagg gccctattct atagtgtcac
      961 ctaaatgcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     1021 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     1081 taataaaatg aggaaattgc atcgcattgt ctga
//
