LOCUS Exported 9706 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS 7860 pRS305-pUG72-HSC82in SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9706) AUTHORS Cowen LE, Lindquist S TITLE Hsp90 potentiates the rapid evolution of new traits: drug resistance in diverse fungi. JOURNAL Science. 2005 Sep 30. 309(5744):2185-9. PUBMED 16195452 REFERENCE 2 (bases 1 to 9706) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9706 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(29..51) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 151..170 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" CDS complement(663..1757) /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" promoter complement(1770..2174) /gene="S. cerevisiae LEU2" /label=LEU2 promoter rep_origin complement(2474..2929) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2611..2632) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2823..2842 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3059..3081 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 3073..3090 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 3074..3090 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 3097..3116 /label=T7 /note="T7 promoter, forward primer" promoter 3097..3115 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 3172..3205 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind 7324..7357 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." primer_bind complement(7400..7416) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7401..7417) /label=pBluescriptKS /note="For pBluescript vector" promoter complement(7446..7464) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(7485..7501) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7485..7501) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(7498..7520) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7509..7525 /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)." promoter complement(7533..7563) /label=lac promoter /note="promoter for the E. coli lac operon" primer_bind complement(7716..7733) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7887..8475) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(8646..9506) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 9269..9288 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(9507..9611) /gene="bla" /label=AmpR promoter primer_bind 9679..9697 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 181 accatatcga ctacgtcgta aggccgtttc tgacagagta aaattcttga gggaactttc 241 accattatgg gaaatggttc aagaaggtat tgacttaaac tccatcaaat ggtcaggtca 301 ttgagtgttt tttatttgtt gtattttttt ttttttagag aaaatcctcc aatatcaaat 361 taggaatcgt agtttcatga ttttctgtta cacctaactt tttgtgtggt gccctcctcc 421 ttgtcaatat taatgttaaa gtgcaattct ttttccttat cacgttgagc cattagtatc 481 aatttgctta cctgtattcc tttactatcc tcctttttct ccttcttgat aaatgtatgt 541 agattgcgta tatagtttcg tctaccctat gaacatattc cattttgtaa tttcgtgtcg 601 tttctattat gaatttcatt tataaagttt atgtacaaat atcataaaaa aagagaatct 661 ttttaagcaa ggattttctt aacttcttcg gcgacagcat caccgacttc ggtggtactg 721 ttggaaccac ctaaatcacc agttctgata cctgcatcca aaaccttttt aactgcatct 781 tcaatggcct taccttcttc aggcaagttc aatgacaatt tcaacatcat tgcagcagac 841 aagatagtgg cgatagggtc aaccttattc tttggcaaat ctggagcaga accgtggcat 901 ggttcgtaca aaccaaatgc ggtgttcttg tctggcaaag aggccaagga cgcagatggc 961 aacaaaccca aggaacctgg gataacggag gcttcatcgg agatgatatc accaaacatg 1021 ttgctggtga ttataatacc atttaggtgg gttgggttct taactaggat catggcggca 1081 gaatcaatca attgatgttg aaccttcaat gtagggaatt cgttcttgat ggtttcctcc 1141 acagtttttc tccataatct tgaagaggcc aaaacattag ctttatccaa ggaccaaata 1201 ggcaatggtg gctcatgttg tagggccatg aaagcggcca ttcttgtgat tctttgcact 1261 tctggaacgg tgtattgttc actatcccaa gcgacaccat caccatcgtc ttcctttctc 1321 ttaccaaagt aaatacctcc cactaattct ctgacaacaa cgaagtcagt acctttagca 1381 aattgtggct tgattggaga taagtctaaa agagagtcgg atgcaaagtt acatggtctt 1441 aagttggcgt acaattgaag ttctttacgg atttttagta aaccttgttc aggtctaaca 1501 ctaccggtac cccatttagg accacccaca gcacctaaca aaacggcatc aaccttcttg 1561 gaggcttcca gcgcctcatc tggaagtggg acacctgtag catcgatagc agcaccacca 1621 attaaatgat tttcgaaatc gaacttgaca ttggaacgaa catcagaaat agctttaaga 1681 accttaatgg cttcggctgt gatttcttga ccaacgtggt cacctggcaa aacgacgatc 1741 ttcttagggg cagacatagg ggcagacatt agaatggtat atccttgaaa tatatatata 1801 tattgctgaa atgtaaaagg taagaaaagt tagaaagtaa gacgattgct aaccacctat 1861 tggaaaaaac aataggtcct taaataatat tgtcaacttc aagtattgtg atgcaagcat 1921 ttagtcatga acgcttctct attctatatg aaaagccggt tccggcctct cacctttcct 1981 ttttctccca atttttcagt tgaaaaaggt atatgcgtca ggcgacctct gaaattaaca 2041 aaaaatttcc agtcatcgaa tttgattctg tgcgatagcg cccctgtgtg ttctcgttat 2101 gttgaggaaa aaaataatgg ttgctaagag attcgaactc ttgcatctta cgatacctga 2161 gtattcccac agttaactgc ggtcaagata tttcttgaat caggcgcctt agaccgctcg 2221 gccaaacaac caattacttg ttgagaaata gagtataatt atcctataaa tataacgttt 2281 ttgaacacac atgaacaagg aagtacagga caattgattt tgaagagaat gtggattttg 2341 atgtaattgt tgggattcca tttttaataa ggcaataata ttaggtatgt ggatatacta 2401 gaagttctcc tcgagggtcg atatgcggtg tgaaataccg cacagatgcg taaggagaaa 2461 ataccgcatc aggaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt 2521 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa 2581 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag 2641 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt 2701 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac 2761 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag 2821 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg 2881 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc gcgccattcg 2941 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc 3001 cagctggcga aggggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc 3061 cagtcacgac gttgtaaaac gacggccagt gaattgtaat acgactcact atagggcgaa 3121 ttggagctcc accgcggccg cataggccac tagtggatct gatatcacct aataacttcg 3181 tatagcatac attatacgaa gttatattaa gggttctcga gagctcccca ttacgcattt 3241 gattataatt tgcttcttag gcaaaattaa tatttacgtt cttttatatt ctttcttttt 3301 gtattcatag aacagcagcc attaccaata gaaagttaaa atagccgccg atgcatttta 3361 ttacccgcct ttctgttttc tgggcacttt tctttctaga aggtgaaaga acaatttttc 3421 tcgttttctc gaacttccac caagcgttgg gtaatgaggg aggagatttg tataaaaaga 3481 gtggcatgtg aactgcctac cgtaagtgac atgaacacat gcattatatt ttttgtgata 3541 tattctttct cttgttttct ttttcttgaa acgctacaga accaatagaa aaatagaatc 3601 attctgaaat atggctggtg aaacttttga atttcaagct gaaatcactc agttgatgag 3661 tttgatcatc aacactgtct attctaacaa ggaaattttc ttgagagaac tgatctctaa 3721 cgcctccgat gctttagaca aaattagata ccaagctttg tctgatccaa agcaattgga 3781 aaccgaacca gatttgttca ttagaatcac cccaaaacca gaagaaaaag ttttggaaat 3841 cagagattct ggtattggta tgaccaaggc tgaattgatt aacaatttgg gtaccattgc 3901 taagtctggt actaaagctt tcatggaagc tctatctgct ggtgccgatg tatccatgat 3961 tggtcaattc ggtgttggtt tttactcttt attcttagtc gccgacagag ttcaagttat 4021 ttccaagaac aatgaggacg aacaatatat ttgggaatct aatgccggtg gttctttcac 4081 cgttactttg gacgaagtta acgaaagaat tggtagaggt accgtcttga gattattctt 4141 gaaagatgac caattggagt acttggaaga aaagagaatt aaagaagtca tcaagagaca 4201 ttctgaattc gttgcttacc ctatccaact tctagtcacc aaggaagtcg aaaaggaagt 4261 tccaattcca gaagaagaaa agaaagacga ggaaaagaag gatgaagatg acaagaaacc 4321 aaaattggaa gaagtcgatg aagaagaaga agaaaagaag ccaaaaacca aaaaagttaa 4381 agaagaggtt caagaattag aagagttgaa caagactaag ccattatgga ctagaaaccc 4441 atctgatatc actcaagagg aatacaatgc tttctataag tctatttcta acgactggga 4501 agacccattg tacgttaagc atttctctgt tgaaggtcaa ttggaattta gagctatctt 4561 gttcattcca aagagagcac cattcgactt atttgagagt aagaagaaga agaacaatat 4621 caagttgtac gttcgtcgtg tcttcatcac tgatgaagct gaagacttga ttccagagtg 4681 gttatctttc gtcaagggtg ttgttgactc tgaagattta ccattgaatt tgtccagaga 4741 aatgttacaa caaaataaga ttatgaaggt tattagaaag aatattgtca agaaattgat 4801 tgaagccttc aacgaaatcg ctgaagactc cgagcaattt gacaaatttt actctgcctt 4861 cgctaagaac attaagctgg gtgtacatga ggacactcaa aacagagctg ctttagctaa 4921 gttgctacgt tacaattcta ctaaatctgt cgatgaattg acttccttga ctgattacgt 4981 tactagaatg ccagaacacc aaaagaacat ctattacatc accggtgaat ctctaaaggc 5041 agtcgaaaag tctccattct tggacgcctt gaaggctaag aactttgaag ttttgttctt 5101 gaccgaccca attgatgaat acgctttcac tcaattgaag gaattcgagg gtaaaacttt 5161 ggttgacatt actaaagatt tcgaattgga agaaacagac gaagaaaaag ctgaaagaga 5221 gaaggagatc aaagaatacg aaccattgac caaggccttg aaggatatct tgggtgacca 5281 agtggagaag gttgttgttt cttacaaatt gctagatgct ccagctgcca tcagaactgg 5341 tcaattcggc tggtctgcta acatggaaag aatcatgaag gctcaagcct tgagagactc 5401 ttccatgtcc tcctacatgt cttccaagaa gactttcgaa atttctccaa aatctccaat 5461 tattaaggaa ttgaaaaaga gagttgatga gggtggtgca caagataaga ccgtcaaaga 5521 tttgactaac ttattattcg agaccgcttt gttgacttct ggtttcagtt tggaagaacc 5581 aacttctttt gcatcaagaa taaatagatt gatttcttta ggtttgaaca ttgatgagga 5641 tgaagaaaca gaaaccgctc cagaagcttc taccgaagct ccagttgaag aggttccagc 5701 tgacaccgag atggaagaag ttgattaatc tcttttttcg ccttcatgtt ttatatatta 5761 tataaatttg tttacttatt tttactattt gtaataatga ttctgcttta cgcgccttta 5821 aaaaagttgg tcttaatgta aacatgaacc atagcacaca atttaagaca tcagctgtca 5881 aatgaccata aaattgtcct accagctgtc cttaaagttt agtgtacact tacgaatgac 5941 ccatggataa actgattatg ttctaaacac tcctcagaag ctcatcgaac tgtcatcctg 6001 cgtgaagatt aaaatccaac ttagaaattt cgagcttacg gagacaatca tatgggagaa 6061 gcaattggaa gatagaaaaa aggtactcgg tacataaata tatgtgattc tgggtagaag 6121 atcggtctgc attggatggt ggtaacgcat ttttttacac acattacttg cctcgagcat 6181 caaatggtgg ttattcgtgg atctatatca cgtgatttgc ttaagaattg tcgttcatgg 6241 tgacactttt agctttgaca tgattaagct catctcaatt gatgttatct aaagtcattt 6301 caactatcta agatgtggtt gtgattgggc cattttgtga aagccagtac gccagcgtca 6361 atacactccc gtcaattagt tgcaccatgt ccacaaaatc atataccagt agagctgaga 6421 ctcatgcaag tccggttgca tcgaaacttt tacgtttaat ggatgaaaag aagaccaatt 6481 tgtgtgcttc tcttgacgtt cgttcgactg atgagctatt gaaacttgtt gaaacgttgg 6541 gtccatacat ttgccttttg aaaacacacg ttgatatctt ggatgatttc agttatgagg 6601 gtactgtcgt tccattgaaa gcattggcag agaaatacaa gttcttgata tttgaggaca 6661 gaaaattcgc cgatatcggt aacacagtca aattacaata tacatcgggc gtttaccgta 6721 tcgcagaatg gtctgatatc accaacgccc acggggttac tggtgctggt attgttgctg 6781 gcttgaaaca aggtgcgcaa gaggtcacca aagaaccaag gggattattg atgcttgctg 6841 aattgtcttc caagggttct ctagcacacg gtgaatatac taagggtacc gttgatattg 6901 caaagagtga taaagatttc gttattgggt tcattgctca gaacgatatg ggaggaagag 6961 aagaagggtt tgattggcta atcatgaccc caggtgtagg tttagacgac aaaggcgatg 7021 cattgggtca gcagtacaga accgtcgacg aagttgtaag tggtggatca gatatcatca 7081 ttgttggcag aggacttttc gccaagggta gagatcctaa ggttgaaggt gaaagataca 7141 gaaatgctgg atgggaagcg taccaaaaga gaatcagcgc tccccattaa ttatacagga 7201 aacttaatag aacaaatcac atatttaatc taatagccac ctgcattggc acggtgcaac 7261 actcacttca acttcatctt acaaaagatc acgtgatctg ttgtattggg atctctagac 7321 ctaataactt cgtatagcat acattatacg aagttatatt aagggttgtc gacctgcagg 7381 aattcgatat caagcttatc gataccgtcg acctcgaggg ggggcccggt acccagcttt 7441 tgttcccttt agtgagggtt aattccgagc ttggcgtaat catggtcata gctgtttcct 7501 gtgtgaaatt gttatccgct cacaattcca cacaacatag gagccggaag cataaagtgt 7561 aaagcctggg gtgcctaatg agtgaggtaa ctcacattaa ttgcgttgcg ctcactgccc 7621 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg 7681 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg 7741 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca 7801 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac 7861 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcg gcccccctga cgagcatcac 7921 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg 7981 ttcccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac 8041 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat 8101 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag 8161 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac 8221 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt 8281 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt 8341 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc 8401 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga 8461 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac 8521 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc 8581 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct 8641 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca 8701 tccatagttg cctgactgcc cgtcgtgtag ataactacga tacgggaggg cttaccatct 8761 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca 8821 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc 8881 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg 8941 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct 9001 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgaaaa 9061 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta 9121 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc 9181 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg 9241 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa 9301 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg 9361 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc 9421 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg 9481 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat 9541 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata 9601 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc 9661 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc //