LOCUS       sequence-251833-        8598 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses thiol redox-sensitive ratiometric sensor roGFP in the 
            mitochondrial intermembrane space of mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8598)
  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 8598)
  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: GPD-roGFP
FEATURES             Location/Qualifiers
     source          1..8598
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..103
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     misc_signal     191..341
                     /label=Ad5 Psi
                     /note="packaging signal for adenovirus serotype 5"
     enhancer        370..673
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        674..877
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     828..848
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1339..1348
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2873..2899
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             join(2900..2902,2903..2905,2906..3619)
                     /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(2945..2966)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3206..3225)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3553..3574
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3664..3686)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      complement(6761..7349)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6841..6860)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7520..8311,8312..8380))
                     /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     8143..8162
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8381..8485)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8553..8571
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 catcatcaat aatatacctt attttggatt gaagccaata tgataatgag ggggtggagt
       61 ttgtgacgtg gcgcggggcg tgggaacggg gcgggtgacg tagtagtgtg gcggaagtgt
      121 gatgttgcaa gtgtggcgga acacatgtaa gcgacggatg tggcaaaagt gacgtttttg
      181 gtgtgcgccg gtgtacacag gaagtgacaa ttttcgcgcg gttttaggcg gatgttgtag
      241 taaatttggg cgtaaccgag taagatttgg ccattttcgc gggaaaactg aataagagga
      301 agtgaaatct gaataatttt gtgttactca tagcgcgtaa tatttgtcta gggagatcag
      361 cctgcaggtc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      421 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      481 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      541 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      601 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      661 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      721 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      781 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      841 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatggt
      901 accgtttaaa ctcgagcggc cgccagtgtg atggatatct gcagaattcc gctcgccgac
      961 ccctggcgtg tgacgaccga tctcaggctg agcaatggcg tttcaaaagg tagtgaaagg
     1021 gactattctt atgggtggag gagctctggc cactgttttg ggactctctc agtttgctca
     1081 ctacagaagg aagcaagtaa gcctggcgta tgtggaagca gcaacatgct tctcagagcc
     1141 cgtgaacagg gagcctcctt ccagagaagc tcagctcatg actctgcaga acacatcgga
     1201 atttgacatc ctcgtcattg gaggaggagc cacaggatgt ggctgtgcgc tagacgccgt
     1261 caccagagga ctgaaaacag cccttgtaga gagaaatgac ttcgcatcag ggactagcag
     1321 cagaagcact aaattgatcc atggtggcgt acgatacctc cagaaggcta tcacgaactt
     1381 ggatgttgaa cagtatagga tggtgaaaga ggcccttcat gaacgtgcca acttactaga
     1441 aatcgctccc catctatcag ctccgctgcc tataatgctg ccactttata agtggtggca
     1501 gttaccttat tactgggtgg gaatcaagat gtatgatctg gttgcaggaa gtcattgcct
     1561 gaagagcagt tatgtcctca gtaaatccag agcccttgag catttcccca tgctccagaa
     1621 ggacaagctg gtcggagcca ttgtctacta tgatggacag cacaatgacg cccggatgaa
     1681 cctcgccatc gccctcaccg ctgccaggta cggggctgcc acggccaatt acatggaggt
     1741 ggtgagcttg ctcaagaaga cagaccccga aactgggaaa gagcgcgtga gtggtgcccg
     1801 gtgcaaggac gtgctcacag ggcatgagtt taacgtgaga gctaaatgcg ttatcaatgc
     1861 caccggccct ttcacagact ccgtgcgcaa aatggatgat aacgatgttg ttcccatctg
     1921 ccagcccagt gcaggggttc acattgtgat gcccgggtac tacagccccg agaacatggg
     1981 acttcttgat cctgcaacca gtgatggcag agttattttc ttcttgcctt gggagaagat
     2041 gacaattgcg ggtaccactg attcgccaac ggacgtcacg caccatccta ttccttcgga
     2101 agacgacatt aacttcatct taaatgaagt gcggaactac ctgagttgcg acgttgaagt
     2161 gagaagaggg gatgtcctgg cagcgtggag tggcattcgc ccgcttgtta ccgatcccaa
     2221 gtctgcaaac actcagtcca tctctcgaaa tcatgttgtg gaagtcagcg acagtggcct
     2281 catcacaata gcaggtggga agtggaccac atatcgctcc atggcagaag atactgtgaa
     2341 caaagctgtc aagcttcaca acttgaatgc aggacccagt aggaccgttg ggctcttcct
     2401 tcaaggaggc aaagattgga gccccacact ctacatcagg cttgtccagg actatgggct
     2461 tgagagtgag gttgcacaac acctggccaa aacctatggt gacaaggctt tcgacgtggc
     2521 caaaatggca agtgtgactg gaaaacggtg gcctgttgtt ggagtgcgtc ttgtgtcaga
     2581 atttccgtac atcgaagcag aggtgaaata tgggattaag gagtacgcat gcaccgctgt
     2641 tgacatgatt tcacgtcgca cccgcctggc ctttcttaat gttcaggctg cagaggaagc
     2701 cctgcccaag attgttgagt taatgggaag agaattgaac tggagtgaat tgaggaaaca
     2761 ggaagaactt gaaacagcca cgaggtttct gtactatgaa atgggctata agtctcgaac
     2821 agaacaactc acagatagta ctgaaatcag cctgctgcct ccagacatcg attatgatgt
     2881 tcctgattat gcaagcttaa tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     2941 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     3001 cgagggcgat gccacctacg gcaagctgac cctgaagttc atcagcacca ccggcaagct
     3061 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     3121 ctaccccgac cacatgaagc ggcacgactt cttcaagtcc gccatgcccg aaggctacgt
     3181 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     3241 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     3301 cggcaacatc ctggggcaca agctggagta caactacaac tgccacaacg tctatatcat
     3361 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     3421 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     3481 gctgctgccc gacaaccact acctgagcac ctgctccgcc ctgagcaaag accccaacga
     3541 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     3601 ggacgagctg tacaagtaag aaggatccgg ctgcctcgcg cgtttcggtg atgacggtga
     3661 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     3721 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat
     3781 gaggtcgact ctagtccccg cggtggcaga tctggaaggt gctgaggtac gatgagaccc
     3841 gcaccaggtg cagaccctgc gagtgtggcg gtaaacatat taggaaccag cctgtgatgc
     3901 tggatgtgac cgaggagctg aggcccgatc acttggtgct ggcctgcacc cgcgctgagt
     3961 ttggctctag cgatgaagat acagattgag gtactgaaat gtgtgggcgt ggcttaaggg
     4021 tgggaaagaa tatataaggt gggggtctta tgtagttttg tatctgtttt gcagcagccg
     4081 ccgccgccat gagcaccaac tcgtttgatg gaagcattgt gagctcatat ttgacaacgc
     4141 gcatgccccc atgggccggg gtgcgtcaga atgtgatggg ctccagcatt gatggtcgcc
     4201 ccgtcctgcc cgcaaactct actaccttga cctacgagac cgtgtctgga acgccgttgg
     4261 agactgcagc ctccgccgcc gcttcagccg ctgcagccac cgcccgcggg attgtgactg
     4321 actttgcttt cctgagcccg cttgcaagca gtgcagcttc ccgttcatcc gcccgcgatg
     4381 acaagttgac ggctcttttg gcacaattgg attctttgac ccgggaactt aatgtcgttt
     4441 ctcagcagct gttggatctg cgccagcagg tttctgccct gaaggcttcc tcccctccca
     4501 atgcggttta aaacataaat aaaaaaccag actctgtttg gatttggatc aagcaagtgt
     4561 cttgctgtct ttatttaggg gttttgcgcg cgcggtaggc ccgggaccag cggtctcggt
     4621 cgttgagggt cctgtgtatt ttttccagga cgtggtaaag gtgactctgg atgttcagat
     4681 acatgggcat aagcccgtct ctggggtgga ggtagcacca ctgcagagct tcatgctgcg
     4741 gggtggtgtt gtagatgatc cagtcgtagc aggagcgctg ggcgtggtgc ctaaaaatgt
     4801 ctttcagtag caagctgatt gccaggggca ggcccttggt gtaagtgttt acaaagcggt
     4861 taagctggga tgggtgcata cgtggggata tgagatgcat cttggactgt atttttaggt
     4921 tggctatgtt cccagccata tccctccggg gattcatgtt gtgcagaacc accagcacag
     4981 tgtatccggt gcacttggga aatttgtcat gtagcttaga aggaaatgcg tggaagaact
     5041 tggagacgcc cttgtgacct ccaagatttt ccatgcattc gtccataatg atggcaatgg
     5101 gcccacgggc ggcggcctgg gcgaagatat ttctgggatc actaacgtca tagttgtgtt
     5161 ccaggatgag atcgtcatag gccattttta caaagcgcgg gcggagggtg ccagactgcg
     5221 gtataatggt tccatccggc ccaggggcgt agttaccctc acagatttgc atttcccacg
     5281 ctttgagttc agatgggggg atcatgtcta cctgcggggc gatgaagaaa acggtttccg
     5341 gggtagggga gatcagctgg gaagaaagca ggttcctgag cagctgcgac ttaccgcagc
     5401 cggtgggccc gtaaatcaca cctattaccg gctgcaactg gtagttaaga gagctgcagc
     5461 tgccgtcatc cctgagcagg ggggccactt cgttaagcat gtccctgact cgcatgtttt
     5521 ccctgaccaa atccgccaga aggcgctcgc cgcccagcga tagcagttct tgcaaggaag
     5581 caaagttttt caacggtttg agaccgtccg ccgtaggcat gcttttgagc gtttgaccaa
     5641 gcagttccag gcggtcccac agctcggtca cctgctctac ggcatctcga tccagcatat
     5701 ctcctcgttt cgcgggttgg ggcggctttc gctgtacggc agtagtcggt gctcgtccag
     5761 acgggccagg gtcatgtctt tccacgggcg cagggtcctc gtcagcgtag tctgggtcac
     5821 ggtgaagggg tgcgctccgg gctgcgcgct ggccagggtg cgcttgaggc tggtcctgct
     5881 ggtgctgaag cgctgccggt cttcgccctg cgcgtcggcc aggtagcatt tgaccatggt
     5941 gtcatagtcc agcccctccg cggcgtggcc cttggcgcgc agcttgccct tggaggaggc
     6001 gccgcacgag gggcagtgca gacttttgag ggcgtagagc ttgggcgcga gaaataccga
     6061 ttccggggag taggcatccg cgccgcaggc cccgcagacg gtctcgcatt ccacgagcca
     6121 ggtgagctct ggccgttcgg ggtcaaaaac caggtttccc ccatgctttt tgatgcgttt
     6181 cttacctctg gtttccatga gccggtgtcc acgctcggtg acgaaaaggc tgtccgtgtc
     6241 cccgtataca gacttgagag gcctgtcctc gaccgatgcc cttgagagcc ttcaacccag
     6301 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
     6361 ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc ggcgaggacc
     6421 gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg
     6481 ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag aagcaggcca
     6541 ttatcgccgg catggcggcc gacgcgctgg gctacgtctt gctggcgttc gcgacgcgag
     6601 gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg atgcccgcgt
     6661 tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt caaggccagc
     6721 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6781 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6841 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6901 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6961 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     7021 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     7081 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     7141 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     7201 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7261 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     7321 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7381 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7441 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7501 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7561 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7621 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7681 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7741 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7801 cagttaatag tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt
     7861 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7921 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7981 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     8041 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     8101 gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata
     8161 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     8221 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     8281 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     8341 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     8401 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8461 aaaataaaca aataggggtt ccgcgcactt ttcggggaaa tgtgccacct gacgtctaag
     8521 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     8581 ttcaattaat taagctag
//
