LOCUS       sequence-475639-        1646 bp    DNA     linear   UNA 07-AUG-2025
DEFINITION  Dual sgRNA targeting TIMM23 expressing dAPEX-BFP targeting to ER.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1646)
  AUTHORS   Thayer JA, Petersen JD, Huang X, Hawrot J, Ramos DM, Sekine S, Li Y,
            Ward ME, Narendra DP
  TITLE     Novel reporter of the PINK1-Parkin mitophagy pathway identifies its 
            damage sensor in the import gate.
  JOURNAL   bioRxiv [Preprint]. 2025 Feb 20:2025.02.19.639160. doi: 
            10.1101/2025.02.19.639160.
  PUBMED    40027798
REFERENCE   2  (bases 1 to 1646)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2025 Feb 20:2025.02.19.639160. doi: 
            10.1101/2025.02.19.639160."
COMMENT     Sequence Label: pLenti_DualsgRNA_TIMM23_ER-dAPEX-BFP2
FEATURES             Location/Qualifiers
     source          1..1646
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      26..35
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     32..94
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTGD"
     CDS             95..841
                     /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="GKSYPTVSADYQKAVEKAKKKLRGFIAEKRCAPLMLRLAFHSAGT
                     FDKGTKTGGPFGTIKHPAELAHSANNGLDIAVRLLEPLKAEFPILSYADFYQLAGVVAV
                     EVTGGPEVPFHPGREDKPEPPPEGRLPDPTKGSDHLRDVFGKAMGLTDQDIVALSGGHT
                     IGAAHKERSGFEGPWTSNPLIFDNSYFTELLSGEKEGLLQLPSDKALLSDPVFRPLVDK
                     YAADEDAFFADYAEAHQKLSELGFADA"
     CDS             896..1606
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
ORIGIN
        1 tccatttcag gtgtcgtgag ctagcgccac catggagaca gacacactcc tgctatgggt
       61 actgctgctc tgggttccag gttccactgg tgacggaaag tcttacccaa ctgtgagtgc
      121 tgattaccag aaggccgttg agaaggcgaa gaagaagctc agaggcttca tcgctgagaa
      181 gagatgcgct cctctaatgc tccgtttggc attccactct gctggaacct ttgacaaggg
      241 cacgaagacc ggtggaccct tcggaaccat caagcaccct gccgaactgg ctcacagcgc
      301 taacaacggt cttgacatcg ctgttaggct tttggagcca ctcaaggcgg agttccctat
      361 tttgagctac gccgatttct accagttggc tggcgttgtt gccgttgagg tcacgggtgg
      421 acctgaagtt ccattccacc ctggaagaga ggacaagcct gagccaccac cagagggtcg
      481 cttgcccgat cccactaagg gttctgacca tttgagagat gtgtttggca aagctatggg
      541 gcttactgac caagatatcg ttgctctatc tgggggtcac actattggag ctgcacacaa
      601 ggagcgttct ggatttgagg gtccctggac ctctaatcct cttattttcg acaactcata
      661 cttcacggag ttgttgagtg gtgagaagga aggtctcctt cagctacctt ctgacaaggc
      721 tcttttgtct gaccctgtat tccgccctct cgttgataaa tatgcagcgg acgaagatgc
      781 cttctttgct gattacgctg aggctcacca aaagctttcc gagcttgggt ttgctgatgc
      841 cggcggtagc agccgtagta gcagcagtgg tggtggtggt agtggtggcg gtggcatggt
      901 gtctaagggc gaagagctga ttaaggagaa catgcacatg aagctgtaca tggagggcac
      961 cgtggacaac catcacttca agtgcacatc cgagggcgaa ggcaagccct acgagggcac
     1021 ccagaccatg agaatcaagg tggtcgaggg cggccctctc cccttcgcct tcgacatcct
     1081 ggctactagc ttcctctacg gcagcaagac cttcatcaac cacacccagg gcatccccga
     1141 cttcttcaag cagtccttcc ctgagggctt cacatgggag agagtcacca catacgaaga
     1201 cgggggcgtg ctgaccgcta cccaggacac cagcctccag gacggctgcc tcatctacaa
     1261 cgtcaagatc agaggggtga acttcacatc caacggccct gtgatgcaga agaaaacact
     1321 cggctgggag gccttcaccg agacgctgta ccccgctgac ggcggcctgg aaggcagaaa
     1381 cgacatggcc ctgaagctcg tgggcgggag ccatctgatc gcaaacgcca agaccacata
     1441 tagatccaag aaacccgcta agaacctcaa gatgcctggc gtctactatg tggactacag
     1501 actggaaaga atcaaggagg ccaacaacga gacctacgtc gagcagcacg aggtggcagt
     1561 ggccagatac tgcgacctcc ctagcaaact ggggcacaag cttaataagg acgagctgta
     1621 agaattcgtc gagggaccta ataact
//
