LOCUS       Exported                7293 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 7293)
  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 7293)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jul 12, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7293
                     /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..1330
                     /locus_tag="ymScarletI3"
                     /label=ymScarletI3
     terminator      1340..1587
                     /gene="S. cerevisiae CYC1"
                     /locus_tag="CYC1 terminator"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1356..1374)
                     /locus_tag="CYC1"
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse
                     primer"
     primer_bind     complement(1605..1624)
                     /locus_tag="T7"
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1606..1624)
                     /locus_tag="T7 promoter"
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1634..1651)
                     /locus_tag="M13 Forward"
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21)
                     Forward"
     primer_bind     complement(1634..1650)
                     /locus_tag="M13 fwd"
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1643..1665)
                     /locus_tag="M13/pUC Forward"
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1795..2250
                     /direction=RIGHT
                     /locus_tag="f1 ori"
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1882..1901)
                     /locus_tag="F1ori-R"
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2092..2113
                     /locus_tag="F1ori-F"
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2550..2954
                     /gene="S. cerevisiae LEU2"
                     /locus_tag="LEU2 promoter"
                     /label=LEU2 promoter
     CDS             2967..4061
                     /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(4554..4573)
                     /locus_tag="pRS-marker"
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     primer_bind     4673..4695
                     /locus_tag="pGEX 3'"
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4733..4751)
                     /locus_tag="pBRforEco"
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     misc_feature    4793..5296
                     /locus_tag="CEN/ARS"
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     promoter        5333..5437
                     /gene="bla"
                     /locus_tag="AmpR promoter"
                     /label=AmpR promoter
     CDS             5438..6298
                     /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(5656..5675)
                     /locus_tag="Amp-R"
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6469..7057
                     /direction=RIGHT
                     /locus_tag="ori"
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6958..6977
                     /locus_tag="pBR322ori-F"
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7211..7228
                     /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 agaatggatt ctactgaagc
      661 tgttattaag gaattcatga gatttaaggt tcatatggaa ggttctatga atggtcatga
      721 atttgaaatt gaaggtgaag gtgaaggtag accatatgaa ggtactcaaa ctgctaagtt
      781 gaaggtcact aagggtggtc cattgccatt ctcttgggat attttgtctc cacaattcat
      841 gtacggttct agagctttta ttaagcatcc agctgacatt ccagattact ggaagcaatc
      901 tttcccagaa ggttttaaat gggaaagagt tatgatcttt gaagatggtg gtactgtttc
      961 tgttactcaa gatacttctt tggaagatgg tactttgatt tacaaggtta agttgagagg
     1021 tggtaacttc ccaccagatg gtccagttat gcaaaagaga actatgggtt gggaagcctc
     1081 tactgaaaga ttgtacccag aagatgttgt tttaaagggt gatattaaga tggctttgag
     1141 attgaaggat ggtggtagat acttggctga ttttaagact acttataagg ctaagaagcc
     1201 agttcaaatg ccaggtgctt tcaacattga tagaaagttg gatattactt ctcataacga
     1261 agattacact gttgttgaac aatacgaaag atctgttgct agacattcta ctggtggttc
     1321 tggtggttct taactcgagt catgtaatta gttatgtcac gcttacattc acgccctccc
     1381 cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt
     1441 tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt
     1501 tttctgtaca gacgcgtgta cgcatgtaac attatactga aaaccttgct tgagaaggtt
     1561 ttgggacgct cgaaggcttt aatttgcggc cggtacccaa ttcgccctat agtgagtcgt
     1621 attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     1681 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     1741 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcgacgcgc
     1801 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     1861 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     1921 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     1981 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     2041 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     2101 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     2161 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     2221 attttaacaa aatattaacg tttacaattt cctgatgcgg tattttctcc ttacgcatct
     2281 gtgcggtatt tcacaccgca tatcgacggt cgaggagaac ttctagtata tccacatacc
     2341 taatattatt gccttattaa aaatggaatc ccaacaatta catcaaaatc cacattctct
     2401 tcaaaatcaa ttgtcctgta cttccttgtt catgtgtgtt caaaaacgtt atatttatag
     2461 gataattata ctctatttct caacaagtaa ttggttgttt ggccgagcgg tctaaggcgc
     2521 ctgattcaag aaatatcttg accgcagtta actgtgggaa tactcaggta tcgtaagatg
     2581 caagagttcg aatctcttag caaccattat ttttttcctc aacataacga gaacacacag
     2641 gggcgctatc gcacagaatc aaattcgatg actggaaatt ttttgttaat ttcagaggtc
     2701 gcctgacgca tatacctttt tcaactgaaa aattgggaga aaaaggaaag gtgagaggcc
     2761 ggaaccggct tttcatatag aatagagaag cgttcatgac taaatgcttg catcacaata
     2821 cttgaagttg acaatattat ttaaggacct attgtttttt ccaataggtg gttagcaatc
     2881 gtcttacttt ctaacttttc ttacctttta catttcagca atatatatat atatttcaag
     2941 gatataccat tctaatgtct gcccctatgt ctgcccctaa gaagatcgtc gttttgccag
     3001 gtgaccacgt tggtcaagaa atcacagccg aagccattaa ggttcttaaa gctatttctg
     3061 atgttcgttc caatgtcaag ttcgatttcg aaaatcattt aattggtggt gctgctatcg
     3121 atgctacagg tgtcccactt ccagatgagg cgctggaagc ctccaagaag gttgatgccg
     3181 ttttgttagg tgctgtgggt ggtcctaaat ggggtaccgg tagtgttaga cctgaacaag
     3241 gtttactaaa aatccgtaaa gaacttcaat tgtacgccaa cttaagacca tgtaactttg
     3301 catccgactc tcttttagac ttatctccaa tcaagccaca atttgctaaa ggtactgact
     3361 tcgttgttgt cagagaatta gtgggaggta tttactttgg taagagaaag gaagacgatg
     3421 gtgatggtgt cgcttgggat agtgaacaat acaccgttcc agaagtgcaa agaatcacaa
     3481 gaatggccgc tttcatggcc ctacaacatg agccaccatt gcctatttgg tccttggata
     3541 aagctaatgt tttggcctct tcaagattat ggagaaaaac tgtggaggaa accatcaaga
     3601 acgaattccc tacattgaag gttcaacatc aattgattga ttctgccgcc atgatcctag
     3661 ttaagaaccc aacccaccta aatggtatta taatcaccag caacatgttt ggtgatatca
     3721 tctccgatga agcctccgtt atcccaggtt ccttgggttt gttgccatct gcgtccttgg
     3781 cctctttgcc agacaagaac accgcatttg gtttgtacga accatgccac ggttctgctc
     3841 cagatttgcc aaagaataag gttgacccta tcgccactat cttgtctgct gcaatgatgt
     3901 tgaaattgtc attgaacttg cctgaagaag gtaaggccat tgaagatgca gttaaaaagg
     3961 ttttggatgc aggtatcaga actggtgatt taggtggttc caacagtacc accgaagtcg
     4021 gtgatgctgt cgccgaagaa gttaagaaaa tccttgctta aaaagattct ctttttttat
     4081 gatatttgta cataaacttt ataaatgaaa ttcataatag aaacgacacg aaattacaaa
     4141 atggaatatg ttcatagggt agacgaaact atatacgcaa tctacataca tttatcaaga
     4201 aggagaaaaa ggaggatagt aaaggaatac aggtaagcaa attgatacta atggctcaac
     4261 gtgataagga aaaagaattg cactttaaca ttaatattga caaggaggag ggcaccacac
     4321 aaaaagttag gtgtaacaga aaatcatgaa actacgattc ctaatttgat attggaggat
     4381 tttctctaaa aaaaaaaaaa tacaacaaat aaaaaacact caatgacctg accatttgat
     4441 ggagtttaag tcaatacctt cttgaaccat ttcccataat ggtgaaagtt ccctcaagaa
     4501 ttttactctg tcagaaacgg ccttacgacg tagtcgatat ggtgcactct cagtacaatc
     4561 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     4621 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     4681 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     4741 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagga cggatcgctt
     4801 gcctgtaact tacacgcgcc tcgtatcttt taatgatgga ataatttggg aatttactct
     4861 gtgtttattt atttttatgt tttgtatttg gattttagaa agtaaataaa gaaggtagaa
     4921 gagttacgga atgaagaaaa aaaaataaac aaaggtttaa aaaatttcaa caaaaagcgt
     4981 actttacata tatatttatt agacaagaaa agcagattaa atagatatac attcgattaa
     5041 cgataagtaa aatgtaaaat cacaggattt tcgtgtgtgg tcttctacac agacaagatg
     5101 aaacaattcg gcattaatac ctgagagcag gaagagcaag ataaaaggta gtatttgttg
     5161 gcgatccccc tagagtcttt tacatcttcg gaaaacaaaa actatttttt ctttaatttc
     5221 tttttttact ttctattttt aatttatata tttatattaa aaaatttaaa ttataattat
     5281 ttttatagca cgtgatgaaa aggacccagg tggcactttt cggggaaatg tgcgcggaac
     5341 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5401 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     5461 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5521 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5581 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     5641 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     5701 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5761 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5821 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5881 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5941 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     6001 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     6061 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     6121 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     6181 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     6241 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     6301 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6361 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6421 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6481 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6541 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6601 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6661 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6721 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6781 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6841 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6901 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6961 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     7021 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     7081 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     7141 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     7201 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     7261 tctccccgcg cgttggccga ttcattaatg cag
//
