LOCUS       sequence-164530-        8213 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The genetically encoded fluorescent H2O2 sensor roGFP2-Tsa2-Delta-CR
            cloned in the yeast p415 vector under the control of a TEF promoter.
            For mitochondrial matrix expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8213)
  AUTHORS   Morgan B, Van Laer K, Owusu TN, Ezerina D, Pastor-Flores D, Amponsah
            PS, Tursch A, Dick TP
  TITLE     Real-time monitoring of basal H2O2 levels with peroxiredoxin-based 
            probes.
  JOURNAL   Nat Chem Biol. 2016 Jun;12(6):437-43. doi: 10.1038/nchembio.2067. 
            Epub 2016 Apr 18.
  PUBMED    27089028
REFERENCE   2  (bases 1 to 8213)
  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: 
            "10.1038/nchembio.2067"; journalName: "Nat Chem Biol"; date: 
            "2016-06"; volume: "12"; issue: "6"; pages: "437-43"
COMMENT     Sequence Label: p415 TEF Su9‐roGFP2‐Tsa2-Delta-CR
FEATURES             Location/Qualifiers
     source          1..8213
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    70..573
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        610..714
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(715..783,784..1575)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(933..952)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1746..2334
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2235..2254
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2488..2505
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(2658..2663,2664..2681,2682..2688)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2696..2712
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2701..2723
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2720..2736
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2720..2736
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2755..2775
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2757..2775
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2793..3191
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     4070..4091
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      4830..5077
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4846..4864)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5095..5114)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5096..5114)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5124..5141)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5124..5140)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5133..5155)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5285..5740
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5372..5391)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5582..5603
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6040..6444
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             6457..7551
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(8044..8063)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8163..8185
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttaggacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata
      121 atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt
      181 aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa
      241 atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata
      301 gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct
      361 tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata
      421 aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact
      481 attttttctt taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa
      541 atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg cacttttcgg
      601 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      661 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
      721 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
      781 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
      841 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
      901 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
      961 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1021 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1081 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1141 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1201 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1261 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1321 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1381 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1441 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1501 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1561 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1621 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1681 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1741 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1801 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1861 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     1921 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1981 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2041 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2101 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2161 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2221 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2281 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2341 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2401 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2461 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     2521 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     2581 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttacctcact
     2641 cattaggcac cccaggcttt acactttatg cttccggctc ctatgttgtg tggaattgtg
     2701 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
     2761 aaccctcact aaagggaaca aaagctggag ctcatagctt caaaatgttt ctactccttt
     2821 tttactcttc cagattttct cggactccgc gcatcgccgt accacttcaa aacacccaag
     2881 cacagcatac taaatttccc ctctttcttc ctctagggtg tcgttaatta cccgtactaa
     2941 aggtttggaa aagaaaaaag agaccgcctc gtttcttttt cttcgtcgaa aaaggcaata
     3001 aaaattttta tcacgtttct ttttcttgaa aatttttttt ttgatttttt tctctttcga
     3061 tgacctccca ttgatattta agttaataaa cggtcttcaa tttctcaagt ttcagtttca
     3121 tttttcttgt tctattacaa ctttttttac ttcttgctca ttagaaagaa agcatagcaa
     3181 tctaatctaa gttttctaga accatggcct ccactcgtgt cctcgcctct cgcctggcct
     3241 cccagatggc tgcttccgcc aaggttgccc gccctgctgt ccgcgttgct caggtcagca
     3301 agcgcaccat ccagactggc tcccccctcc agaccctcaa gcgcacccag atgacctcca
     3361 tcgtcaacgc caccacccgc caggctttcc agaagcgcgc ctactcttcc gctagcgaat
     3421 tctcaaaggg tgaagaattg tttacaggtg ttgttcctat tttagtcgaa ttggacggtg
     3481 acgttaatgg tcataagttt agtgttagtg gtgaaggtga aggtgacgca acatacggta
     3541 aattgacctt gaagtttatt tcaactactg gtaaattgcc agttccttgg ccaactttgg
     3601 taaccacttt aacatatggt gtacaatgtt tcagtagata ccctgatcat atgaaacaac
     3661 acgacttttt caagtctgct atgccagaag gttacgttca agaaagaact attttcttta
     3721 aggatgacgg taactacaag accagagcag aagtcaaatt tgaaggtgac actttggtta
     3781 acagaatcga attgaagggt atcgatttca aggaagacgg taacatcttg ggtcataaat
     3841 tggaatacaa ctacaactgt cacaatgtat acataatggc cgataagcaa aagaatggta
     3901 tcaaagtcaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     3961 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     4021 tgagcacctg ctccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     4081 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagactagtg
     4141 gtggttcagg tggtggtggt tcaggtggtg gtggttcagg tggaggagga tcaggaggag
     4201 gaggatcagg aggaggagga tcaggaggag aattcgtagc agaagttcaa aaacaagccc
     4261 caccatttaa gaaaaccgcc gtagtcgacg gtatcttcga ggaaatttca ctggaaaagt
     4321 ataaaggtaa gtacgttgtt ctagcttttg tcccattggc tttttcattt gtctgtccaa
     4381 ctgagattgt tgcgttttcc gatgccgcca agaaattcga agatcagggc gcccaagttt
     4441 tatttgcctc caccgactct gaatattcct tactggcatg gaccaacctt cccagaaaag
     4501 acggtggatt aggtccagtt aaagttcctt tgcttgctga taagaatcat tccttatcca
     4561 gagactatgg cgttttgatt gaaaaagaag gtatagcttt aagaggtttg ttcataatcg
     4621 acccgaaggg aatcattaga catatcacta tcaatgattt atctgttggc agaaacgtca
     4681 atgaagcttt gagattagtc gaaggtttcc agtggactga caaaaatggt acagttttgc
     4741 cagccaactg gaccccagga gccgccacca tcaaacctga cgttaaagat tccaaggagt
     4801 atttcaaaaa tgccaataat taactcgagt catgtaatta gttatgtcac gcttacattc
     4861 acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta
     4921 ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat
     4981 ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga aaaccttgct
     5041 tgagaaggtt ttgggacgct cgaaggcttt aatttgcggc cggtacccaa ttcgccctat
     5101 agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     5161 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     5221 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     5281 cgcgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     5341 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     5401 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     5461 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     5521 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     5581 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     5641 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     5701 tttaacgcga attttaacaa aatattaacg tttacaattt cctgatgcgg tattttctcc
     5761 ttacgcatct gtgcggtatt tcacaccgca tatcgacggt cgaggagaac ttctagtata
     5821 tccacatacc taatattatt gccttattaa aaatggaatc ccaacaatta catcaaaatc
     5881 cacattctct tcaaaatcaa ttgtcctgta cttccttgtt catgtgtgtt caaaaacgtt
     5941 atatttatag gataattata ctctatttct caacaagtaa ttggttgttt ggccgagcgg
     6001 tctaaggcgc ctgattcaag aaatatcttg accgcagtta actgtgggaa tactcaggta
     6061 tcgtaagatg caagagttcg aatctcttag caaccattat ttttttcctc aacataacga
     6121 gaacacacag gggcgctatc gcacagaatc aaattcgatg actggaaatt ttttgttaat
     6181 ttcagaggtc gcctgacgca tatacctttt tcaactgaaa aattgggaga aaaaggaaag
     6241 gtgagaggcc ggaaccggct tttcatatag aatagagaag cgttcatgac taaatgcttg
     6301 catcacaata cttgaagttg acaatattat ttaaggacct attgtttttt ccaataggtg
     6361 gttagcaatc gtcttacttt ctaacttttc ttacctttta catttcagca atatatatat
     6421 atatttcaag gatataccat tctaatgtct gcccctatgt ctgcccctaa gaagatcgtc
     6481 gttttgccag gtgaccacgt tggtcaagaa atcacagccg aagccattaa ggttcttaaa
     6541 gctatttctg atgttcgttc caatgtcaag ttcgatttcg aaaatcattt aattggtggt
     6601 gctgctatcg atgctacagg tgtcccactt ccagatgagg cgctggaagc ctccaagaag
     6661 gttgatgccg ttttgttagg tgctgtgggt ggtcctaaat ggggtaccgg tagtgttaga
     6721 cctgaacaag gtttactaaa aatccgtaaa gaacttcaat tgtacgccaa cttaagacca
     6781 tgtaactttg catccgactc tcttttagac ttatctccaa tcaagccaca atttgctaaa
     6841 ggtactgact tcgttgttgt cagagaatta gtgggaggta tttactttgg taagagaaag
     6901 gaagacgatg gtgatggtgt cgcttgggat agtgaacaat acaccgttcc agaagtgcaa
     6961 agaatcacaa gaatggccgc tttcatggcc ctacaacatg agccaccatt gcctatttgg
     7021 tccttggata aagctaatgt tttggcctct tcaagattat ggagaaaaac tgtggaggaa
     7081 accatcaaga acgaattccc tacattgaag gttcaacatc aattgattga ttctgccgcc
     7141 atgatcctag ttaagaaccc aacccaccta aatggtatta taatcaccag caacatgttt
     7201 ggtgatatca tctccgatga agcctccgtt atcccaggtt ccttgggttt gttgccatct
     7261 gcgtccttgg cctctttgcc agacaagaac accgcatttg gtttgtacga accatgccac
     7321 ggttctgctc cagatttgcc aaagaataag gttgacccta tcgccactat cttgtctgct
     7381 gcaatgatgt tgaaattgtc attgaacttg cctgaagaag gtaaggccat tgaagatgca
     7441 gttaaaaagg ttttggatgc aggtatcaga actggtgatt taggtggttc caacagtacc
     7501 accgaagtcg gtgatgctgt cgccgaagaa gttaagaaaa tccttgctta aaaagattct
     7561 ctttttttat gatatttgta cataaacttt ataaatgaaa ttcataatag aaacgacacg
     7621 aaattacaaa atggaatatg ttcatagggt agacgaaact atatacgcaa tctacataca
     7681 tttatcaaga aggagaaaaa ggaggatagt aaaggaatac aggtaagcaa attgatacta
     7741 atggctcaac gtgataagga aaaagaattg cactttaaca ttaatattga caaggaggag
     7801 ggcaccacac aaaaagttag gtgtaacaga aaatcatgaa actacgattc ctaatttgat
     7861 attggaggat tttctctaaa aaaaaaaaaa tacaacaaat aaaaaacact caatgacctg
     7921 accatttgat ggagtttaag tcaatacctt cttgaaccat ttcccataat ggtgaaagtt
     7981 ccctcaagaa ttttactctg tcagaaacgg ccttacgacg tagtcgatat ggtgcactct
     8041 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     8101 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     8161 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cga
//
