LOCUS       sequence-202531-        1898 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian expression plasmid to target the rmRuby1-roGFP1 sensor to 
            the mitochondrial matrix..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1898)
  AUTHORS   Norcross S, Trull KJ, Snaider J, Doan S, Tat K, Huang L, Tantama M
  TITLE     Extending roGFP Emission via Forster-Type Resonance Energy Transfer 
            Relay Enables Simultaneous Dual Compartment Ratiometric Redox 
            Imaging in Live Cells.
  JOURNAL   ACS Sens. 2017 Nov 22;2(11):1721-1729. doi: 
            10.1021/acssensors.7b00689. Epub 2017 Nov 7.
  PUBMED    29072071
REFERENCE   2  (bases 1 to 1898)
  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"; doi: 
            "10.1021/acssensors.7b00689"; journalName: "ACS Sens"; date: 
            "2017-11-22- 22"; volume: "2"; issue: "11"; pages: "1721-1729"
COMMENT     Sequence Label: GW1-mito-mRuby2-roGFP1
FEATURES             Location/Qualifiers
     source          1..1898
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             41..115
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             137..211
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             233..307
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             329..403
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             443..1153
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     CDS             join(1175..1177,1178..1891)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FISTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNCHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTCSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1217..1238)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1478..1497)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1825..1846
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 ccatcggcgg agcaggtgtg cctgaggcct ctagaccacc atgagcgtgc tgacaccatt
       61 gttgctcaga ggcctcactg gttcagccag gcggctgcca gtcccaagag ctaagatcca
      121 tagtctggga gatccaatgt cagttctcac tccccttttg cttaggggac tgaccggaag
      181 cgctcggcgg ttgcctgtcc cacgcgccaa gattcactcc ctgggcgacc ctatgagtgt
      241 ccttactcct cttttgctga gaggcttgac agggagcgct cggaggctcc cagtgcctcg
      301 cgcaaagatc cattctctcg gagatcccat gtccgtcctc actcctctgc tgctgcgcgg
      361 gcttacaggt tccgctcgcc gcctgcctgt tcctcgggca aagatacaca gtctgggcga
      421 tcctcaccgg cgggctgctg ctatggtgtc taagggcgaa gagctgatca aggaaaatat
      481 gcgtatgaag gtggtcatgg aaggttcggt caacggccac caattcaaat gcacaggtga
      541 aggagaaggc aatccgtaca tgggaactca aaccatgagg atcaaagtca tcgagggagg
      601 acccctgcca tttgcctttg acattcttgc cacgtcgttc atgtatggca gccgtacttt
      661 tatcaagtac ccgaaaggca ttcctgattt ctttaaacag tcctttcctg agggttttac
      721 ttgggaaaga gttacgagat acgaagatgg tggagtcgtc accgtcatgc aggacaccag
      781 ccttgaggat ggctgtctcg tttaccacgt ccaagtcaga ggggtaaact ttccctccaa
      841 tggtcccgtg atgcagaaga agaccaaggg ttgggagcct aatacagaga tgatgtatcc
      901 agcagatggt ggtctgaggg gatacactca tatggcactg aaagttgatg gtggtggcca
      961 tctgtcttgc tctttcgtaa caacttacag gtcaaaaaag accgtcggga acatcaagat
     1021 gcccggtatc catgccgttg atcaccgcct ggaaaggtta gaggaaagtg acaatgaaat
     1081 gttcgtagta caacgcgaac acgcagttgc caagttcgcc gggcttggtg gtgggatgga
     1141 cgagctgtac aagggaggtt caggaggtcg atccgtgagc aagggcgagg agctgttcac
     1201 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     1261 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctccac
     1321 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc actctgtctt acggcgtgca
     1381 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     1441 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     1501 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     1561 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca actgccacaa
     1621 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     1681 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     1741 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acctgctccg ccctgagcaa
     1801 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     1861 cactctcggc atggacgagc tgtacaagta atcgacga
//
