LOCUS Exported 7781 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Yeast expression plasmid for roGFP2-Grx2; yeast codon optimized roGFP; yeast Grx2. ACCESSION . VERSION . KEYWORDS p415TEF cyto roGFP2-Grx2(yeast) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7781) AUTHORS Morgan B, Ezerina D, Amoako TN, Riemer J, Seedorf M, Dick TP TITLE Multiple glutathione disulfide removal pathways mediate cytosolic redox homeostasis. JOURNAL Nat Chem Biol. 2013 Feb;9(2):119-25. doi: 10.1038/nchembio.1142. Epub 2012 Dec 16. PUBMED 23242256 REFERENCE 2 (bases 1 to 7781) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7781 /organism="synthetic DNA construct" /mol_type="other DNA" 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 715..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 2658..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 3195..3211 /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind 3195..3211 /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(4379..4395) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4380..4396) /label=pBluescriptKS /note="For pBluescript vector" terminator 4398..4645 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind complement(4414..4432) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" primer_bind complement(4663..4682) /label=T7 /note="T7 promoter, forward primer" promoter complement(4664..4682) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4692..4709) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4692..4708) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4701..4723) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 4853..5308 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4940..4959) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5150..5171 /label=F1ori-F /note="F1 origin, forward primer" promoter 5608..6012 /gene="S. cerevisiae LEU2" /label=LEU2 promoter CDS 6025..7119 /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(7612..7631) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 7731..7753 /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 actagtggat ccaccatggc tagcgagttc tcaaagggtg 3241 aagaattgtt tacaggtgtt gttcctattt tagtcgaatt ggacggtgac gttaatggtc 3301 ataagtttag tgttagtggt gaaggtgaag gtgacgcaac atacggtaaa ttgaccttga 3361 agtttatttc aactactggt aaattgccag ttccttggcc aactttggta accactttaa 3421 catatggtgt acaatgtttc agtagatacc ctgatcatat gaaacaacac gactttttca 3481 agtctgctat gccagaaggt tacgttcaag aaagaactat tttctttaag gatgacggta 3541 actacaagac cagagcagaa gtcaaatttg aaggtgacac tttggttaac agaatcgaat 3601 tgaagggtat cgatttcaag gaagacggta acatcttggg tcataaattg gaatacaact 3661 acaactgtca caatgtatac ataatggccg ataagcaaaa gaatggtatc aaagtcaact 3721 tcaagatcag acataacatc gaagatggtt ctgttcaatt agctgaccac tatcaacaaa 3781 atacacctat tggtgacggt cctgttttgt taccagacaa ccattacttg tctacctgct 3841 cagctttatc caaagatcca aatgaaaaga gagaccatat ggtattgtta gaatttgtca 3901 ccgcagcagg tatcacattg ggtatggatg aattgtatac tagtggtggt tcaggtggtg 3961 gtggttcagg tggtggtggt tcaggtggag gaggatcagg aggaggagga tcaggaggag 4021 gaggatcagg aggagaattc atggtatccc aggaaacagt tgctcacgta aaggatctga 4081 ttggccaaaa ggaagtgttt gttgcagcaa agacatactg cccttactgt aaagctactt 4141 tgtctaccct cttccaagaa ttgaacgttc ccaaatccaa ggcccttgtg ttggaattag 4201 atgaaatgag caatggctca gagattcaag acgctttaga agaaatctcg ggccaaaaaa 4261 ctgtacctaa cgtatacatc aatggcaagc acattggtgg taacagcgat ttggaaactt 4321 tgaagaaaaa tggcaagtta gctgaaatat tgaagccggt atttcaatag aagcttatcg 4381 ataccgtcga cctcgagtca tgtaattagt tatgtcacgc ttacattcac gccctccccc 4441 cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta 4501 tttttttata gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt 4561 tctgtacaga cgcgtgtacg catgtaacat tatactgaaa accttgcttg agaaggtttt 4621 gggacgctcg aaggctttaa tttgcggccg gtacccaatt cgccctatag tgagtcgtat 4681 tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc 4741 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc 4801 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cgacgcgccc 4861 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt 4921 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc 4981 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta 5041 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc 5101 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg 5161 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt 5221 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat 5281 tttaacaaaa tattaacgtt tacaatttcc tgatgcggta ttttctcctt acgcatctgt 5341 gcggtatttc acaccgcata tcgacggtcg aggagaactt ctagtatatc cacataccta 5401 atattattgc cttattaaaa atggaatccc aacaattaca tcaaaatcca cattctcttc 5461 aaaatcaatt gtcctgtact tccttgttca tgtgtgttca aaaacgttat atttatagga 5521 taattatact ctatttctca acaagtaatt ggttgtttgg ccgagcggtc taaggcgcct 5581 gattcaagaa atatcttgac cgcagttaac tgtgggaata ctcaggtatc gtaagatgca 5641 agagttcgaa tctcttagca accattattt ttttcctcaa cataacgaga acacacaggg 5701 gcgctatcgc acagaatcaa attcgatgac tggaaatttt ttgttaattt cagaggtcgc 5761 ctgacgcata tacctttttc aactgaaaaa ttgggagaaa aaggaaaggt gagaggccgg 5821 aaccggcttt tcatatagaa tagagaagcg ttcatgacta aatgcttgca tcacaatact 5881 tgaagttgac aatattattt aaggacctat tgttttttcc aataggtggt tagcaatcgt 5941 cttactttct aacttttctt accttttaca tttcagcaat atatatatat atttcaagga 6001 tataccattc taatgtctgc ccctatgtct gcccctaaga agatcgtcgt tttgccaggt 6061 gaccacgttg gtcaagaaat cacagccgaa gccattaagg ttcttaaagc tatttctgat 6121 gttcgttcca atgtcaagtt cgatttcgaa aatcatttaa ttggtggtgc tgctatcgat 6181 gctacaggtg tcccacttcc agatgaggcg ctggaagcct ccaagaaggt tgatgccgtt 6241 ttgttaggtg ctgtgggtgg tcctaaatgg ggtaccggta gtgttagacc tgaacaaggt 6301 ttactaaaaa tccgtaaaga acttcaattg tacgccaact taagaccatg taactttgca 6361 tccgactctc ttttagactt atctccaatc aagccacaat ttgctaaagg tactgacttc 6421 gttgttgtca gagaattagt gggaggtatt tactttggta agagaaagga agacgatggt 6481 gatggtgtcg cttgggatag tgaacaatac accgttccag aagtgcaaag aatcacaaga 6541 atggccgctt tcatggccct acaacatgag ccaccattgc ctatttggtc cttggataaa 6601 gctaatgttt tggcctcttc aagattatgg agaaaaactg tggaggaaac catcaagaac 6661 gaattcccta cattgaaggt tcaacatcaa ttgattgatt ctgccgccat gatcctagtt 6721 aagaacccaa cccacctaaa tggtattata atcaccagca acatgtttgg tgatatcatc 6781 tccgatgaag cctccgttat cccaggttcc ttgggtttgt tgccatctgc gtccttggcc 6841 tctttgccag acaagaacac cgcatttggt ttgtacgaac catgccacgg ttctgctcca 6901 gatttgccaa agaataaggt tgaccctatc gccactatct tgtctgctgc aatgatgttg 6961 aaattgtcat tgaacttgcc tgaagaaggt aaggccattg aagatgcagt taaaaaggtt 7021 ttggatgcag gtatcagaac tggtgattta ggtggttcca acagtaccac cgaagtcggt 7081 gatgctgtcg ccgaagaagt taagaaaatc cttgcttaaa aagattctct ttttttatga 7141 tatttgtaca taaactttat aaatgaaatt cataatagaa acgacacgaa attacaaaat 7201 ggaatatgtt catagggtag acgaaactat atacgcaatc tacatacatt tatcaagaag 7261 gagaaaaagg aggatagtaa aggaatacag gtaagcaaat tgatactaat ggctcaacgt 7321 gataaggaaa aagaattgca ctttaacatt aatattgaca aggaggaggg caccacacaa 7381 aaagttaggt gtaacagaaa atcatgaaac tacgattcct aatttgatat tggaggattt 7441 tctctaaaaa aaaaaaaata caacaaataa aaaacactca atgacctgac catttgatgg 7501 agtttaagtc aataccttct tgaaccattt cccataatgg tgaaagttcc ctcaagaatt 7561 ttactctgtc agaaacggcc ttacgacgta gtcgatatgg tgcactctca gtacaatctg 7621 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg 7681 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg 7741 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg a //