LOCUS       Exported                7538 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast plasmid with Hsp82.
ACCESSION   .
VERSION     .
KEYWORDS    p414TEF_Hsp82_CycTER
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7538)
  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 7538)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7538
                     /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"
     CDS             3216..3233
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      5392..5639
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5408..5426)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5657..5676)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5658..5676)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5686..5703)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5686..5702)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5695..5717)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5843..6298
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5930..5949)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6140..6161
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6397..7071)
                     /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(7072..7352)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(7369..7388)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7488..7510
                     /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 actagtatgg gtggtcacca ccatcaccac cacggtggtc
     3241 atatggctag tgaaactttt gaatttcaag ctgaaattac tcagttgatg agtttgatca
     3301 tcaacaccgt ctattctaac aaggaaattt tcttgagaga actgatatct aatgcctcgg
     3361 atgcgttgga taaaattaga tacaaatctt tgtctgatcc aaagcaattg gaaacagaac
     3421 cagatctctt tattagaatc actccaaagc cagagcaaaa agttttggaa atcagagatt
     3481 ctggtattgg tatgaccaag gctgaattga ttaataactt gggtaccatt gccaagtctg
     3541 gtaccaaagc cttcatggaa gctctatctg ctggtgccga tgtatccatg attggtcaat
     3601 tcggtgttgg tttttactct ttattcttag ttgccgacag agttcaggtt atttcaaaga
     3661 gcaacgacga cgaacaatac atctgggaat ccaacgctgg tggttctttc actgttactc
     3721 tagacgaagt taatgaaaga attggtaggg gtaccatctt gaggttattc ttgaaagatg
     3781 accaattgga gtacttggaa gaaaagagaa taaaggaagt tatcaagaga cattctgagt
     3841 tcgtggccta cccaatccaa ttagtcgtca ccaaggaagt tgaaaaggaa gttccaattc
     3901 cagaagaaga aaagaaagac gaggaaaaga aggatgagga aaagaaggat gaagacgaca
     3961 agaaaccaaa actcgaggaa gtcgatgaag aagaggaaaa gaagccaaag acgaaaaaag
     4021 ttaaagaaga agttcaggag atagaagaac taaacaagac taagcctttg tggactagaa
     4081 acccatctga tatcactcaa gaagaataca atgctttcta taagtctatt tcaaacgact
     4141 gggaagaccc attgtacgtt aagcatttct ccgttgaagg tcaattggaa tttagagcta
     4201 tcttattcat tccaaagaga gcaccattcg acttgtttga gagtaaaaag aagaagaata
     4261 atatcaagtt gtacgttcgt cgtgttttca tcactgatga agctgaagac ttgattccag
     4321 agtggttatc tttcgtcaag ggtgttgttg actctgagga tttaccattg aatttgtcca
     4381 gagaaatgtt acaacaaaat aagatcatga aggttattag aaagaacatt gtcaaaaagt
     4441 tgattgaagc cttcaacgaa attgctgaag actctgaaca atttgaaaag ttctactcgg
     4501 ctttctccaa aaatatcaag ttgggtgtac atgaagatac ccaaaacagg gctgctttgg
     4561 ctaagttgtt acgttacaac tctaccaagt ccgtagatga gttgacttcc ttaactgatt
     4621 acgttaccag aatgccagaa caccaaaaga acatctacta catcactggt gaatctctaa
     4681 aggctgtcga aaagtctcca tttttggatg ccttgaaggc taaaaacttc gaggttttgt
     4741 tcttgaccga cccaattgat gaatacgcct tcactcaatt gaaggaattc gaaggtaaaa
     4801 ctttggttga cattactaaa gatttcgaat tggaagaaac tgacgaagaa aaagctgaaa
     4861 gagagaagga gatcaaagaa tatgaaccat tgaccaaagc tttgaaagaa attttgggtg
     4921 accaagtgga gaaagttgtt gtttcttaca aattgttgga tgccccagct gctatcagaa
     4981 ctggtcaatt tggttggtct gctaacatgg aaagaatcat gaaggctcaa gccttgagag
     5041 actcttccat gtcctcctac atgtcttcca agaagacttt cgaaatttct ccaaaatctc
     5101 caattatcaa ggaattgaaa aagagagttg acgaaggtgg tgctcaagac aagactgtca
     5161 aggacttgac taagttatta tatgaaactg ctttgttgac ttccggcttc agtttggacg
     5221 aaccaacttc ctttgcatca agaattaaca gattgatctc tttaggcctg aacattgatg
     5281 aggatgaaga aacagagact gctccagaag catccaccgc agctccggtt gaagaggttc
     5341 cagctgacac cgaaatggaa gaggtagatt aatggatcca atcgattcga gtcatgtaat
     5401 tagttatgtc acgcttacat tcacgccctc cccccacatc cgctctaacc gaaaaggaag
     5461 gagttagaca acctgaagtc taggtcccta tttatttttt tatagttatg ttagtattaa
     5521 gaacgttatt tatatttcaa atttttcttt tttttctgta cagacgcgtg tacgcatgta
     5581 acattatact gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct ttaatttgcg
     5641 gccggtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt
     5701 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     5761 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     5821 gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     5881 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     5941 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     6001 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     6061 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     6121 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     6181 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     6241 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttcc
     6301 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata ggcaagtgca
     6361 caaacaatac ttaaataaat actactcagt aataacctat ttcttagcat ttttgacgaa
     6421 atttgctatt ttgttagagt cttttacacc atttgtctcc acacctccgc ttacatcaac
     6481 accaataacg ccatttaatc taagcgcatc accaacattt tctggcgtca gtccaccagc
     6541 taacataaaa tgtaagcttt cggggctctc ttgccttcca acccagtcag aaatcgagtt
     6601 ccaatccaaa agttcacctg tcccacctgc ttctgaatca aacaagggaa taaacgaatg
     6661 aggtttctgt gaagctgcac tgagtagtat gttgcagtct tttggaaata cgagtctttt
     6721 aataactggc aaaccgagga actcttggta ttcttgccac gactcatctc catgcagttg
     6781 gacgatatca atgccgtaat cattgaccag agccaaaaca tcctccttag gttgattacg
     6841 aaacacgcca accaagtatt tcggagtgcc tgaactattt ttatatgctt ttacaagact
     6901 tgaaattttc cttgcaataa ccgggtcaat tgttctcttt ctattgggca cacatataat
     6961 acccagcaag tcagcatcgg aatctagagc acattctgcg gcctctgtgc tctgcaagcc
     7021 gcaaactttc accaatggac cagaactacc tgtgaaatta ataacagaca tactccaagc
     7081 tgcctttgtg tgcttaatca cgtatactca cgtgctcaat agtcaccaat gccctccctc
     7141 ttggccctct ccttttcttt tttcgaccga attaattctt aatcggcaaa aaaagaaaag
     7201 ctccggatca agattgtacg taaggtgaca agctattttt caataaagaa tatcttccac
     7261 tactgccatc tggcgtcata actgcaaagt acacatatat tacgatgctg tctattaaat
     7321 gcttcctata ttatatatat agtaatgtcg tttatggtgc actctcagta caatctgctc
     7381 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     7441 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     7501 gtgtcagagg ttttcaccgt catcaccgaa acgcgcga
//
