LOCUS       Exported                7536 bp ds-DNA     circular SYN 15-MAY-2024
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 7536)
  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 7536)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jul 12, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7536
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        88..118
                     /locus_tag="lac promoter"
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    126..142
                     /locus_tag="lac operator"
                     /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     131..153
                     /locus_tag="M13/pUC Reverse"
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     150..166
                     /locus_tag="M13 rev"
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     150..166
                     /locus_tag="M13 Reverse"
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     185..205
                     /locus_tag="T3"
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        187..205
                     /locus_tag="T3 promoter"
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        223..621
                     /gene="S. cerevisiae TEF1"
                     /locus_tag="TEF1 promoter"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     misc_feature    644..1573
                     /locus_tag="yemiRFP670"
                     /label=yemiRFP670
     terminator      1583..1830
                     /gene="S. cerevisiae CYC1"
                     /locus_tag="CYC1 terminator"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1599..1617)
                     /locus_tag="CYC1"
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse
                     primer"
     primer_bind     complement(1848..1867)
                     /locus_tag="T7"
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1849..1867)
                     /locus_tag="T7 promoter"
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1877..1894)
                     /locus_tag="M13 Forward"
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21)
                     Forward"
     primer_bind     complement(1877..1893)
                     /locus_tag="M13 fwd"
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1886..1908)
                     /locus_tag="M13/pUC Forward"
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2038..2493
                     /direction=RIGHT
                     /locus_tag="f1 ori"
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2125..2144)
                     /locus_tag="F1ori-R"
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2335..2356
                     /locus_tag="F1ori-F"
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2793..3197
                     /gene="S. cerevisiae LEU2"
                     /locus_tag="LEU2 promoter"
                     /label=LEU2 promoter
     CDS             3210..4304
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /locus_tag="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(4797..4816)
                     /locus_tag="pRS-marker"
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     primer_bind     4916..4938
                     /locus_tag="pGEX 3'"
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4976..4994)
                     /locus_tag="pBRforEco"
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     misc_feature    5036..5539
                     /locus_tag="CEN/ARS"
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     promoter        5576..5680
                     /gene="bla"
                     /locus_tag="AmpR promoter"
                     /label=AmpR promoter
     CDS             5681..6541
                     /codon_start=1
                     /gene="bla"
                     /locus_tag="AmpR"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5899..5918)
                     /locus_tag="Amp-R"
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6712..7300
                     /direction=RIGHT
                     /locus_tag="ori"
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7201..7220
                     /locus_tag="pBR322ori-F"
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7454..7471
                     /locus_tag="L4440"
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
       61 ttacctcact cattaggcac cccaggcttt acactttatg cttccggctc ctatgttgtg
      121 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
      181 gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctcatagctt caaaatgttt
      241 ctactccttt tttactcttc cagattttct cggactccgc gcatcgccgt accacttcaa
      301 aacacccaag cacagcatac taaatttccc ctctttcttc ctctagggtg tcgttaatta
      361 cccgtactaa aggtttggaa aagaaaaaag agaccgcctc gtttcttttt cttcgtcgaa
      421 aaaggcaata aaaattttta tcacgtttct ttttcttgaa aatttttttt ttgatttttt
      481 tctctttcga tgacctccca ttgatattta agttaataaa cggtcttcaa tttctcaagt
      541 ttcagtttca tttttcttgt tctattacaa ctttttttac ttcttgctca ttagaaagaa
      601 agcatagcaa tctaatctaa gttttctaga accatggtct agaatggctg aaggttctgt
      661 tgctagacaa ccagatttgt tgacttgtga acatgaagaa attcatttgg ctggttctat
      721 tcaaccacat ggtgctttgt tggtcgtttc tgaacatgac catagagtta ttcaagcctc
      781 tgctaatgcc gctgaattct taaacttggg ttctgtttta ggtgttccat tagctgaaat
      841 tgatggtgat ttgttaatta agattttacc acatttagat ccaaccgctg aaggtatgcc
      901 agttgctgtt agatgtagaa tcggtaatcc ttccactgaa tactgtggtt tgatgcatag
      961 accaccagaa ggtggtttga ttattgaatt ggaaagagct ggtccatcta ttgatttatc
     1021 cggtactttg gctccagctt tggaaagaat tagaactgct ggttctttaa gagctttgtg
     1081 tgatgacact gttttgttat tccaacaatg tactggttat gatagagtta tggtttacag
     1141 attcgatgaa caaggtcatg gtttagtttt ctctgaatgt catgtccctg gtttggaatc
     1201 ttacttcggt aatagatacc catcctctac tgttccacaa atggctagac aattatacgt
     1261 tagacaaaga gttagagttt tggtcgatgt tacttaccaa ccagttccat tggaaccaag
     1321 attatcccca ttgactggta gagatttaga tatgtctggt tgtttcttaa gatctatgtc
     1381 tccttgtcat ttgcaattct tgaaggatat gggtgttaga gctactttgg ccgtttcttt
     1441 agttgttggt ggtaagttgt ggggtttggt tgtttgtcac cattacttgc caagattcat
     1501 tagattcgaa ttaagagcta tctgtaagag attggctgaa agaattgcta ctagaattac
     1561 cgctttagaa tcttaactcg agtcatgtaa ttagttatgt cacgcttaca ttcacgccct
     1621 ccccccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct
     1681 atttattttt ttatagttat gttagtatta agaacgttat ttatatttca aatttttctt
     1741 ttttttctgt acagacgcgt gtacgcatgt aacattatac tgaaaacctt gcttgagaag
     1801 gttttgggac gctcgaaggc tttaatttgc ggccggtacc caattcgccc tatagtgagt
     1861 cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     1921 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     1981 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcgacg
     2041 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2101 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     2161 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     2221 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     2281 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     2341 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     2401 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     2461 cgaattttaa caaaatatta acgtttacaa tttcctgatg cggtattttc tccttacgca
     2521 tctgtgcggt atttcacacc gcatatcgac ggtcgaggag aacttctagt atatccacat
     2581 acctaatatt attgccttat taaaaatgga atcccaacaa ttacatcaaa atccacattc
     2641 tcttcaaaat caattgtcct gtacttcctt gttcatgtgt gttcaaaaac gttatattta
     2701 taggataatt atactctatt tctcaacaag taattggttg tttggccgag cggtctaagg
     2761 cgcctgattc aagaaatatc ttgaccgcag ttaactgtgg gaatactcag gtatcgtaag
     2821 atgcaagagt tcgaatctct tagcaaccat tatttttttc ctcaacataa cgagaacaca
     2881 caggggcgct atcgcacaga atcaaattcg atgactggaa attttttgtt aatttcagag
     2941 gtcgcctgac gcatatacct ttttcaactg aaaaattggg agaaaaagga aaggtgagag
     3001 gccggaaccg gcttttcata tagaatagag aagcgttcat gactaaatgc ttgcatcaca
     3061 atacttgaag ttgacaatat tatttaagga cctattgttt tttccaatag gtggttagca
     3121 atcgtcttac tttctaactt ttcttacctt ttacatttca gcaatatata tatatatttc
     3181 aaggatatac cattctaatg tctgccccta tgtctgcccc taagaagatc gtcgttttgc
     3241 caggtgacca cgttggtcaa gaaatcacag ccgaagccat taaggttctt aaagctattt
     3301 ctgatgttcg ttccaatgtc aagttcgatt tcgaaaatca tttaattggt ggtgctgcta
     3361 tcgatgctac aggtgtccca cttccagatg aggcgctgga agcctccaag aaggttgatg
     3421 ccgttttgtt aggtgctgtg ggtggtccta aatggggtac cggtagtgtt agacctgaac
     3481 aaggtttact aaaaatccgt aaagaacttc aattgtacgc caacttaaga ccatgtaact
     3541 ttgcatccga ctctctttta gacttatctc caatcaagcc acaatttgct aaaggtactg
     3601 acttcgttgt tgtcagagaa ttagtgggag gtatttactt tggtaagaga aaggaagacg
     3661 atggtgatgg tgtcgcttgg gatagtgaac aatacaccgt tccagaagtg caaagaatca
     3721 caagaatggc cgctttcatg gccctacaac atgagccacc attgcctatt tggtccttgg
     3781 ataaagctaa tgttttggcc tcttcaagat tatggagaaa aactgtggag gaaaccatca
     3841 agaacgaatt ccctacattg aaggttcaac atcaattgat tgattctgcc gccatgatcc
     3901 tagttaagaa cccaacccac ctaaatggta ttataatcac cagcaacatg tttggtgata
     3961 tcatctccga tgaagcctcc gttatcccag gttccttggg tttgttgcca tctgcgtcct
     4021 tggcctcttt gccagacaag aacaccgcat ttggtttgta cgaaccatgc cacggttctg
     4081 ctccagattt gccaaagaat aaggttgacc ctatcgccac tatcttgtct gctgcaatga
     4141 tgttgaaatt gtcattgaac ttgcctgaag aaggtaaggc cattgaagat gcagttaaaa
     4201 aggttttgga tgcaggtatc agaactggtg atttaggtgg ttccaacagt accaccgaag
     4261 tcggtgatgc tgtcgccgaa gaagttaaga aaatccttgc ttaaaaagat tctctttttt
     4321 tatgatattt gtacataaac tttataaatg aaattcataa tagaaacgac acgaaattac
     4381 aaaatggaat atgttcatag ggtagacgaa actatatacg caatctacat acatttatca
     4441 agaaggagaa aaaggaggat agtaaaggaa tacaggtaag caaattgata ctaatggctc
     4501 aacgtgataa ggaaaaagaa ttgcacttta acattaatat tgacaaggag gagggcacca
     4561 cacaaaaagt taggtgtaac agaaaatcat gaaactacga ttcctaattt gatattggag
     4621 gattttctct aaaaaaaaaa aaatacaaca aataaaaaac actcaatgac ctgaccattt
     4681 gatggagttt aagtcaatac cttcttgaac catttcccat aatggtgaaa gttccctcaa
     4741 gaattttact ctgtcagaaa cggccttacg acgtagtcga tatggtgcac tctcagtaca
     4801 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     4861 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     4921 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     4981 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta ggacggatcg
     5041 cttgcctgta acttacacgc gcctcgtatc ttttaatgat ggaataattt gggaatttac
     5101 tctgtgttta tttattttta tgttttgtat ttggatttta gaaagtaaat aaagaaggta
     5161 gaagagttac ggaatgaaga aaaaaaaata aacaaaggtt taaaaaattt caacaaaaag
     5221 cgtactttac atatatattt attagacaag aaaagcagat taaatagata tacattcgat
     5281 taacgataag taaaatgtaa aatcacagga ttttcgtgtg tggtcttcta cacagacaag
     5341 atgaaacaat tcggcattaa tacctgagag caggaagagc aagataaaag gtagtatttg
     5401 ttggcgatcc ccctagagtc ttttacatct tcggaaaaca aaaactattt tttctttaat
     5461 ttcttttttt actttctatt tttaatttat atatttatat taaaaaattt aaattataat
     5521 tatttttata gcacgtgatg aaaaggaccc aggtggcact tttcggggaa atgtgcgcgg
     5581 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5641 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5701 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5761 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5821 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5881 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5941 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     6001 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     6061 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     6121 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     6181 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     6241 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     6301 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     6361 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     6421 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6481 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6541 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     6601 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6661 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6721 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6781 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6841 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6901 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6961 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     7021 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     7081 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     7141 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     7201 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     7261 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     7321 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     7381 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     7441 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     7501 gcctctcccc gcgcgttggc cgattcatta atgcag
//
