LOCUS       Exported                8681 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  yeast plasmid with Hsp82.
ACCESSION   .
VERSION     .
KEYWORDS    p414ADH_Hsp82_CycTER
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8681)
  AUTHORS   Jiang L, Mishra P, Hietpas RT, Zeldovich KB, Bolon DN
  TITLE     Latent effects of Hsp90 mutants revealed at reduced expression 
            levels.
  JOURNAL   PLoS Genet. 2013 Jun;9(6):e1003600. doi: 
            10.1371/journal.pgen.1003600. Epub 2013 Jun 27.
  PUBMED    23825969
REFERENCE   2  (bases 1 to 8681)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8681
                     /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        3577..4281
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             4316..4333
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6512..6528)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6513..6529)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      6531..6778
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6547..6565)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(6796..6815)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6797..6815)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6825..6842)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6825..6841)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6834..6856)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6986..7441
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7073..7092)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7283..7304
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(7540..8214)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(8215..8495)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(8512..8531)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8631..8653
                     /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 ctctaaaaca agaagagggt tgactacatc
     2821 acgatgaggg ggatcgaaga aatgatggta aatgaaatag gaaatcaagg agcatgaagg
     2881 caaaagacaa atataagggt cgaacgaaaa ataaagtgaa aagtgttgat atgatgtatt
     2941 tggctttgcg gcgccgaaaa aacgagttta cgcaattgca caatcatgct gactctgtgg
     3001 cggacccgcg ctcttgccgg cccggcgata acgctgggcg tgaggctgtg cccggcggag
     3061 ttttttgcgc ctgcattttc caaggtttac cctgcgctaa ggggcgagat tggagaagca
     3121 ataagaatgc cggttggggt tgcgatgatg acgaccacga caactggtgt cattatttaa
     3181 gttgccgaaa gaacctgagt gcatttgcaa catgagtata ctagaagaat gagccaagac
     3241 ttgcgagacg cgagtttgcc ggtggtgcga acaatagagc gaccatgacc ttgaaggtga
     3301 gacgcgcata accgctagag tactttgaag aggaaacagc aatagggttg ctaccagtat
     3361 aaatagacag gtacatacaa cactggaaat ggttgtctgt ttgagtacgc tttcaattca
     3421 tttgggtgtg cactttatta tgttacaata tggaagggaa ctttacactt ctcctatgca
     3481 catatattaa ttaaagtcca atgctagtag agaagggggg taacacccct ccgcgctctt
     3541 ttccgatttt tttctaaacc gtggaatatt tcggatatcc ttttgttgtt tccgggtgta
     3601 caatatggac ttcctctttt ctggcaacca aacccataca tcgggattcc tataatacct
     3661 tcgttggtct ccctaacatg taggtggcgg aggggagata tacaatagaa cagataccag
     3721 acaagacata atgggctaaa caagactaca ccaattacac tgcctcattg atggtggtac
     3781 ataacgaact aatactgtag ccctagactt gatagccatc atcatatcga agtttcacta
     3841 ccctttttcc atttgccatc tattgaagta ataataggcg catgcaactt cttttctttt
     3901 tttttctttt ctctctcccc cgttgttgtc tcaccatatc cgcaatgaca aaaaaatgat
     3961 ggaagacact aaaggaaaaa attaacgaca aagacagcac caacagatgt cgttgttcca
     4021 gagctgatga ggggtatctc gaagcacacg aaactttttc cttccttcat tcacgcacac
     4081 tactctctaa tgagcaacgg tatacggcct tccttccagt tacttgaatt tgaaataaaa
     4141 aaaagtttgc tgtcttgcta tcaagtataa atagacctgc aattattaat cttttgtttc
     4201 ctcgtcattg ttctcgttcc ctttcttcct tgtttctttt tctgcacaat atttcaagct
     4261 ataccaagca tacaatcaac tccaagctgg ccgctctaga actagtatgg gtggtcacca
     4321 ccatcaccac cacggtggtc atatggctag tgaaactttt gaatttcaag ctgaaattac
     4381 tcagttgatg agtttgatca tcaacaccgt ctattctaac aaggaaattt tcttgagaga
     4441 actgatatct aatgcctcgg atgcgttgga taaaattaga tacaaatctt tgtctgatcc
     4501 aaagcaattg gaaacagaac cagatctctt tattagaatc actccaaagc cagagcaaaa
     4561 agttttggaa atcagagatt ctggtattgg tatgaccaag gctgaattga ttaataactt
     4621 gggtaccatt gccaagtctg gtaccaaagc cttcatggaa gctctatctg ctggtgccga
     4681 tgtatccatg attggtcaat tcggtgttgg tttttactct ttattcttag ttgccgacag
     4741 agttcaggtt atttcaaaga gcaacgacga cgaacaatac atctgggaat ccaacgctgg
     4801 tggttctttc actgttactc tagacgaagt taatgaaaga attggtaggg gtaccatctt
     4861 gaggttattc ttgaaagatg accaattgga gtacttggaa gaaaagagaa taaaggaagt
     4921 tatcaagaga cattctgagt tcgtggccta cccaatccaa ttagtcgtca ccaaggaagt
     4981 tgaaaaggaa gttccaattc cagaagaaga aaagaaagac gaggaaaaga aggatgagga
     5041 aaagaaggat gaagacgaca agaaaccaaa actcgaggaa gtcgatgaag aagaggaaaa
     5101 gaagccaaag acgaaaaaag ttaaagaaga agttcaagag atagaagaac taaacaagac
     5161 taagcctttg tggactagaa acccatctga tatcactcaa gaagaataca atgctttcta
     5221 taagtctatt tcaaacgact gggaagaccc attgtacgtt aagcatttct ccgttgaagg
     5281 tcaattggaa tttagagcta tcttattcat tccaaagaga gcaccattcg acttgtttga
     5341 gagtaaaaag aagaagaata atatcaagtt gtacgttcgt cgtgttttca tcactgatga
     5401 agctgaagac ttgattccag agtggttatc tttcgtcaag ggtgttgttg actctgagga
     5461 tttaccattg aatttgtcca gagaaatgtt acaacaaaat aagatcatga aggttattag
     5521 aaagaacatt gtcaaaaagt tgattgaagc cttcaacgaa attgctgaag actctgaaca
     5581 atttgaaaag ttctactcgg ctttctccaa aaatatcaag ttgggtgtac atgaagatac
     5641 ccaaaacagg gctgctttgg ctaagttgtt acgttacaac tctaccaagt ccgtagatga
     5701 gttgacttcc ttaactgatt acgttaccag aatgccagaa caccaaaaga acatctacta
     5761 catcactggt gaatctctaa aggctgtcga aaagtctcca tttttggatg ccttgaaggc
     5821 taaaaacttc gaggttttgt tcttgaccga cccaattgat gaatacgcct tcactcaatt
     5881 gaaggaattc gaaggtaaaa ctttggttga cattactaaa gatttcgaat tggaagaaac
     5941 tgacgaagaa aaagctgaaa gagagaagga gatcaaagaa tatgaaccat tgaccaaagc
     6001 tttgaaagaa attttgggtg accaagtgga gaaagttgtt gtttcttaca aattgttgga
     6061 tgccccagct gctatcagaa ctggtcaatt tggttggtct gctaacatgg aaagaatcat
     6121 gaaggctcaa gccttgagag actcttccat gtcctcctac atgtcttcca agaagacttt
     6181 cgaaatttct ccaaaatctc caattatcaa ggaattgaaa aagagagttg acgaaggtgg
     6241 tgctcaagac aagactgtca aggacttgac taagttatta tatgaaactg ctttgttgac
     6301 ttccggcttc agtttggacg aaccaacttc ctttgcatca agaattaaca gattgatctc
     6361 tttaggcctg aacattgatg aggatgaaga aacagagact gctccagaag catccaccgc
     6421 agctccggtt gaagaggttc cagctgacac cgaaatggaa gaggtagatt aatggatccc
     6481 ccgggctgca ggaattcgat atcaagctta tcgataccgt cgacctcgag tcatgtaatt
     6541 agttatgtca cgcttacatt cacgccctcc ccccacatcc gctctaaccg aaaaggaagg
     6601 agttagacaa cctgaagtct aggtccctat ttattttttt atagttatgt tagtattaag
     6661 aacgttattt atatttcaaa tttttctttt ttttctgtac agacgcgtgt acgcatgtaa
     6721 cattatactg aaaaccttgc ttgagaaggt tttgggacgc tcgaaggctt taatttgcgg
     6781 ccggtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta
     6841 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     6901 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     6961 cgcagcctga atggcgaatg gcgcgacgcg ccctgtagcg gcgcattaag cgcggcgggt
     7021 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     7081 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     7141 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     7201 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     7261 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     7321 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     7381 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     7441 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc ataggcaagt
     7501 gcacaaacaa tacttaaata aatactactc agtaataacc tatttcttag catttttgac
     7561 gaaatttgct attttgttag agtcttttac accatttgtc tccacacctc cgcttacatc
     7621 aacaccaata acgccattta atctaagcgc atcaccaaca ttttctggcg tcagtccacc
     7681 agctaacata aaatgtaagc tttcggggct ctcttgcctt ccaacccagt cagaaatcga
     7741 gttccaatcc aaaagttcac ctgtcccacc tgcttctgaa tcaaacaagg gaataaacga
     7801 atgaggtttc tgtgaagctg cactgagtag tatgttgcag tcttttggaa atacgagtct
     7861 tttaataact ggcaaaccga ggaactcttg gtattcttgc cacgactcat ctccatgcag
     7921 ttggacgata tcaatgccgt aatcattgac cagagccaaa acatcctcct taggttgatt
     7981 acgaaacacg ccaaccaagt atttcggagt gcctgaacta tttttatatg cttttacaag
     8041 acttgaaatt ttccttgcaa taaccgggtc aattgttctc tttctattgg gcacacatat
     8101 aatacccagc aagtcagcat cggaatctag agcacattct gcggcctctg tgctctgcaa
     8161 gccgcaaact ttcaccaatg gaccagaact acctgtgaaa ttaataacag acatactcca
     8221 agctgccttt gtgtgcttaa tcacgtatac tcacgtgctc aatagtcacc aatgccctcc
     8281 ctcttggccc tctccttttc ttttttcgac cgaattaatt cttaatcggc aaaaaaagaa
     8341 aagctccgga tcaagattgt acgtaaggtg acaagctatt tttcaataaa gaatatcttc
     8401 cactactgcc atctggcgtc ataactgcaa agtacacata tattacgatg ctgtctatta
     8461 aatgcttcct atattatata tatagtaatg tcgtttatgg tgcactctca gtacaatctg
     8521 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     8581 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     8641 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg a
//
