LOCUS       sequence-145885-        1064 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses APEX2 in mitochondria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1064)
  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 1064)
  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: mito-V5-APEX2
FEATURES             Location/Qualifiers
     source          1..1064
                     /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"
     CDS             186..227
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             228..974
                     /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"
     promoter        complement(1008..1026)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1009..1026)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1046..1063)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
ORIGIN
        1 ctatataagc agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat
       61 taatacgact cactataggg agacccaagc ttgcggccgc caccatgctg gccacccgcg
      121 tgttcagcct ggtgggcaag cgcgccatca gcaccagcgt gtgcgtgcgc gcccacaagg
      181 atccaggcaa gcccatcccc aaccccctgc tgggcctgga cagcaccgga aagtcttacc
      241 caactgtgag tgctgattac caggacgccg ttgagaaggc gaagaagaag ctcagaggct
      301 tcatcgctga gaagagatgc gctcctctaa tgctccgttt ggcattccac tctgctggaa
      361 cctttgacaa gggcacgaag accggtggac ccttcggaac catcaagcac cctgccgaac
      421 tggctcacag cgctaacaac ggtcttgaca tcgctgttag gcttttggag ccactcaagg
      481 cggagttccc tattttgagc tacgccgatt tctaccagtt ggctggcgtt gttgccgttg
      541 aggtcacggg tggacctaag gttccattcc accctggaag agaggacaag cctgagccac
      601 caccagaggg tcgcttgcct gatcccacta agggttctga ccatttgaga gatgtgtttg
      661 gcaaagctat ggggcttact gaccaagata tcgttgctct atctgggggt cacactattg
      721 gagctgcaca caaggagcgt tctggatttg agggtccctg gacctctaat cctcttattt
      781 tcgacaactc atacttcacg gagttgttga gtggtgagaa ggaaggtctc cttcagctac
      841 cttctgacaa ggctcttttg tctgaccctg tattccgccc tctcgttgat aaatatgcag
      901 cggacgaaga tgccttcttt gctgattacg ctgaggctca ccaaaagctt tccgagcttg
      961 ggtttgctga tgcctaatga ctcgagcatg catctagagg gccctattct atagtgtcac
     1021 ctaaatgcta gagctcgctg atcagcctcg actgtgcctt ctag
//
