LOCUS       sequence-369148-        1119 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Lentiviral conditional expression of mitochondrial-targeted eGFP in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1119)
  AUTHORS   Michaelis JB, Bozkurt S, Schafer JA, Munch C
  TITLE     Monitoring Mitochondrial Protein Import Using Mitochondrial 
            Targeting Sequence (MTS)-eGFP.
  JOURNAL   Bio Protoc. 2022 Dec 20;12(24):e4578. doi: 10.21769/BioProtoc.4578. 
            eCollection 2022 Dec 20.
  PUBMED    36618095
REFERENCE   2  (bases 1 to 1119)
  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: "Bio 
            Protoc."; date: "2022-12-20"; volume: "12(24):e4578. doi"; pages: " 
            10.21769/BioProtoc.4578. eCollection 2022 Dec 20"
COMMENT     Sequence Label: pLD-puro-MTS-EGFP
FEATURES             Location/Qualifiers
     source          1..1119
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      220..229
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(226..228,229..231,232..945)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(226..228,229..231,232..942)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(271..292)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(532..551)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     879..900
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             963..980
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 accatggcct ccactcgtgt cctcgcctct cgcctggcct cccggatggc tgcttccgcc
       61 aaggttgccc gccctgctgt ccgcgttgct caggtcagca agcgcaccat ccagactggc
      121 tcccccctcc agaccctcaa gcgcacccag atgacctcca tcgtcaacgc caccacccgc
      181 caggctttcc agaagcgcgc ctactcttcc ggatccatcg ccaccatggt gagcaagggc
      241 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
      301 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
      361 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
      421 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
      481 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
      541 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
      601 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
      661 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
      721 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
      781 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
      841 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
      901 accgccgccg ggatcactct cggcatggac gagctgtaca agtaattagt aatgaaccgg
      961 tccaccacca ccaccaccac taaggatccg gggttggggt tgcgcctttt ccaaggcagc
     1021 cctgggtttg cgcagggacg cggctgctct gggcgtggtt ccgggaaacg cagcggcgcc
     1081 gaccctgggt ctcgcacatt cttcacgtcc gttcgcagg
//
