LOCUS       sequence-257693-        6929 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses thiol redox-sensitive ratiometric sensor roGFP in the 
            mitochondrial matrix of mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6929)
  AUTHORS   Waypa GB, Marks JD, Guzy R, Mungai PT, Schriewer J, Dokic D, 
            Schumacker PT
  TITLE     Hypoxia triggers subcellular compartmental redox signaling in 
            vascular smooth muscle cells.
  JOURNAL   Circ Res. 2010 Feb 19;106(3):526-35. doi: 
            10.1161/CIRCRESAHA.109.206334. Epub 2009 Dec 17.
  PUBMED    20019331
REFERENCE   2  (bases 1 to 6929)
  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: 
            "10.1161/CIRCRESAHA.109.206334"; journalName: "Circ Res"; date: 
            "2010-02-19- 19"; volume: "106"; issue: "3"; pages: "526-35"
COMMENT     Sequence Label: Matrix-roGFP
FEATURES             Location/Qualifiers
     source          1..6929
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        12..315
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        316..519
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     470..490
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(600..602,603..605,606..1319)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFISTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNCHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTCSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(645..666)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(906..925)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1253..1274
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1360..1494
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1384..1403)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1438..1457
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1637..1659)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      complement(4734..5322)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4814..4833)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5493..6284,6285..6353))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6116..6135
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6354..6458)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6526..6544
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     repeat_region   6572..6674
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     misc_signal     6762..6912
                     /label=Ad5 Psi
                     /note="packaging signal for adenovirus serotype 5"
ORIGIN
        1 agcctgcagg tcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
       61 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      121 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      181 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      241 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      301 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      361 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      421 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      481 ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatg
      541 gtaccgccac catgctgagc ctgcgccaga gcatccgctt cttcaagcgc agcggcatca
      601 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
      661 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
      721 gcaagctgac cctgaagttc atcagcacca ccggcaagct gcccgtgccc tggcccaccc
      781 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
      841 ggcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
      901 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
      961 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     1021 agctggagta caactacaac tgccacaacg tctatatcat ggccgacaag cagaagaacg
     1081 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     1141 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     1201 acctgagcac ctgctccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     1261 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaac
     1321 tcgagcggaa gggcgaattc gcggccgcca cagcggggag atccagacat gataagatac
     1381 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     1441 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     1501 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     1561 aagtaaaacc tctacaaatg tggtatggct gattatgatc cggctgcctc gcgcgtttcg
     1621 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     1681 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     1741 ggggcgcagc catgaggtcg actctagtcc ccgcggtggc agatctggaa ggtgctgagg
     1801 tacgatgaga cccgcaccag gtgcagaccc tgcgagtgtg gcggtaaaca tattaggaac
     1861 cagcctgtga tgctggatgt gaccgaggag ctgaggcccg atcacttggt gctggcctgc
     1921 acccgcgctg agtttggctc tagcgatgaa gatacagatt gaggtactga aatgtgtggg
     1981 cgtggcttaa gggtgggaaa gaatatataa ggtgggggtc ttatgtagtt ttgtatctgt
     2041 tttgcagcag ccgccgccgc catgagcacc aactcgtttg atggaagcat tgtgagctca
     2101 tatttgacaa cgcgcatgcc cccatgggcc ggggtgcgtc agaatgtgat gggctccagc
     2161 attgatggtc gccccgtcct gcccgcaaac tctactacct tgacctacga gaccgtgtct
     2221 ggaacgccgt tggagactgc agcctccgcc gccgcttcag ccgctgcagc caccgcccgc
     2281 gggattgtga ctgactttgc tttcctgagc ccgcttgcaa gcagtgcagc ttcccgttca
     2341 tccgcccgcg atgacaagtt gacggctctt ttggcacaat tggattcttt gacccgggaa
     2401 cttaatgtcg tttctcagca gctgttggat ctgcgccagc aggtttctgc cctgaaggct
     2461 tcctcccctc ccaatgcggt ttaaaacata aataaaaaac cagactctgt ttggatttgg
     2521 atcaagcaag tgtcttgctg tctttattta ggggttttgc gcgcgcggta ggcccgggac
     2581 cagcggtctc ggtcgttgag ggtcctgtgt attttttcca ggacgtggta aaggtgactc
     2641 tggatgttca gatacatggg cataagcccg tctctggggt ggaggtagca ccactgcaga
     2701 gcttcatgct gcggggtggt gttgtagatg atccagtcgt agcaggagcg ctgggcgtgg
     2761 tgcctaaaaa tgtctttcag tagcaagctg attgccaggg gcaggccctt ggtgtaagtg
     2821 tttacaaagc ggttaagctg ggatgggtgc atacgtgggg atatgagatg catcttggac
     2881 tgtattttta ggttggctat gttcccagcc atatccctcc ggggattcat gttgtgcaga
     2941 accaccagca cagtgtatcc ggtgcacttg ggaaatttgt catgtagctt agaaggaaat
     3001 gcgtggaaga acttggagac gcccttgtga cctccaagat tttccatgca ttcgtccata
     3061 atgatggcaa tgggcccacg ggcggcggcc tgggcgaaga tatttctggg atcactaacg
     3121 tcatagttgt gttccaggat gagatcgtca taggccattt ttacaaagcg cgggcggagg
     3181 gtgccagact gcggtataat ggttccatcc ggcccagggg cgtagttacc ctcacagatt
     3241 tgcatttccc acgctttgag ttcagatggg gggatcatgt ctacctgcgg ggcgatgaag
     3301 aaaacggttt ccggggtagg ggagatcagc tgggaagaaa gcaggttcct gagcagctgc
     3361 gacttaccgc agccggtggg cccgtaaatc acacctatta ccggctgcaa ctggtagtta
     3421 agagagctgc agctgccgtc atccctgagc aggggggcca cttcgttaag catgtccctg
     3481 actcgcatgt tttccctgac caaatccgcc agaaggcgct cgccgcccag cgatagcagt
     3541 tcttgcaagg aagcaaagtt tttcaacggt ttgagaccgt ccgccgtagg catgcttttg
     3601 agcgtttgac caagcagttc caggcggtcc cacagctcgg tcacctgctc tacggcatct
     3661 cgatccagca tatctcctcg tttcgcgggt tggggcggct ttcgctgtac ggcagtagtc
     3721 ggtgctcgtc cagacgggcc agggtcatgt ctttccacgg gcgcagggtc ctcgtcagcg
     3781 tagtctgggt cacggtgaag gggtgcgctc cgggctgcgc gctggccagg gtgcgcttga
     3841 ggctggtcct gctggtgctg aagcgctgcc ggtcttcgcc ctgcgcgtcg gccaggtagc
     3901 atttgaccat ggtgtcatag tccagcccct ccgcggcgtg gcccttggcg cgcagcttgc
     3961 ccttggagga ggcgccgcac gaggggcagt gcagactttt gagggcgtag agcttgggcg
     4021 cgagaaatac cgattccggg gagtaggcat ccgcgccgca ggccccgcag acggtctcgc
     4081 attccacgag ccaggtgagc tctggccgtt cggggtcaaa aaccaggttt cccccatgct
     4141 ttttgatgcg tttcttacct ctggtttcca tgagccggtg tccacgctcg gtgacgaaaa
     4201 ggctgtccgt gtccccgtat acagacttga gaggcctgtc ctcgaccgat gcccttgaga
     4261 gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
     4321 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt
     4381 ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc
     4441 ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa acgtttcggc
     4501 gagaagcagg ccattatcgc cggcatggcg gccgacgcgc tgggctacgt cttgctggcg
     4561 ttcgcgacgc gaggctggat ggccttcccc attatgattc ttctcgcttc cggcggcatc
     4621 gggatgcccg cgttgcaggc catgctgtcc aggcaggtag atgacgacca tcagggacag
     4681 cttcaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4741 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4801 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4861 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4921 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4981 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5041 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5101 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5161 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5221 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5281 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5341 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5401 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5461 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5521 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5581 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5641 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5701 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5761 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg
     5821 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     5881 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     5941 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6001 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6061 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat
     6121 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6181 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6241 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6301 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6361 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6421 gaatgtattt agaaaaataa acaaataggg gttccgcgca cttttcgggg aaatgtgcca
     6481 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     6541 aggccctttc gtcttcaatt aattaagcta gcatcatcaa taatatacct tattttggat
     6601 tgaagccaat atgataatga gggggtggag tttgtgacgt ggcgcggggc gtgggaacgg
     6661 ggcgggtgac gtagtagtgt ggcggaagtg tgatgttgca agtgtggcgg aacacatgta
     6721 agcgacggat gtggcaaaag tgacgttttt ggtgtgcgcc ggtgtacaca ggaagtgaca
     6781 attttcgcgc ggttttaggc ggatgttgta gtaaatttgg gcgtaaccga gtaagatttg
     6841 gccattttcg cgggaaaact gaataagagg aagtgaaatc tgaataattt tgtgttactc
     6901 atagcgcgta atatttgtct agggagatc
//
