LOCUS       Exported                7182 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  yeast plasmid with Hsp82.
ACCESSION   .
VERSION     .
KEYWORDS    p414CYC_Hsp82_noTER
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7182)
  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 7182)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7182
                     /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"
     CDS             3104..3121
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5297..5316)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5298..5316)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5326..5343)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5326..5342)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5335..5357)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5487..5942
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5574..5593)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5784..5805
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6041..6715)
                     /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(6716..6996)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(7013..7032)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7132..7154
                     /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 ctcatttggc gagcgttggt tggtggatca
     2821 agcccacgcg taggcaatcc tcgagcagat ccgccaggcg tgtatatata gcgtggatgg
     2881 ccaggcaact ttagtgctga cacatacagg catatatata tgtgtgcgac gacacatgat
     2941 catatggcat tcatgtgctc tgtatgtata taaaactctt gttttcttct tttctctaaa
     3001 tattctttcc ttatacatta ggacctttgc agcataaatt actatacttc tatagacacg
     3061 caaacacaaa tacacacact aatctagaac tagtatgggt ggtcaccacc atcaccacca
     3121 cggtggtcat atggctagtg aaacttttga atttcaagct gaaattactc agttgatgag
     3181 tttgatcatc aacaccgtct attctaacaa ggaaattttc ttgagagaac tgatatctaa
     3241 tgcctcggat gcgttggata aaattagata caaatctttg tctgatccaa agcaattgga
     3301 aacagaacca gatctcttta ttagaatcac tccaaagcca gagcaaaaag ttttggaaat
     3361 cagagattct ggtattggta tgaccaaggc tgaattgatt aataacttgg gtaccattgc
     3421 caagtctggt accaaagcct tcatggaagc tctatctgct ggtgccgatg tatccatgat
     3481 tggtcaattc ggtgttggtt tttactcttt attcttagtt gccgacagag ttcaggttat
     3541 ttcaaagagc aacgacgacg aacaatacat ctgggaatcc aacgctggtg gttctttcac
     3601 tgttactcta gacgaagtta atgaaagaat tggtaggggt accatcttga ggttattctt
     3661 gaaagatgac caattggagt acttggaaga aaagagaata aaggaagtta tcaagagaca
     3721 ttctgagttc gtggcctacc caatccaatt agtcgtcacc aaggaagttg aaaaggaagt
     3781 tccaattcca gaagaagaaa agaaagacga ggaaaagaag gatgaggaaa agaaggatga
     3841 agacgacaag aaaccaaaac tcgaggaagt cgatgaagaa gaggaaaaga agccaaagac
     3901 gaaaaaagtt aaagaagaag ttcaagagat agaagaacta aacaagacta agcctttgtg
     3961 gactagaaac ccatctgata tcactcaaga agaatacaat gctttctata agtctatttc
     4021 aaacgactgg gaagacccat tgtacgttaa gcatttctcc gttgaaggtc aattggaatt
     4081 tagagctatc ttattcattc caaagagagc accattcgac ttgtttgaga gtaaaaagaa
     4141 gaagaataat atcaagttgt acgttcgtcg tgttttcatc actgatgaag ctgaagactt
     4201 gattccagag tggttatctt tcgtcaaggg tgttgttgac tctgaggatt taccattgaa
     4261 tttgtccaga gaaatgttac aacaaaataa gatcatgaag gttattagaa agaacattgt
     4321 caaaaagttg attgaagcct tcaacgaaat tgctgaagac tctgaacaat ttgaaaagtt
     4381 ctactcggct ttctccaaaa atatcaagtt gggtgtacat gaagataccc aaaacagggc
     4441 tgctttggct aagttgttac gttacaactc taccaagtcc gtagatgagt tgacttcctt
     4501 aactgattac gttaccagaa tgccagaaca ccaaaagaac atctactaca tcactggtga
     4561 atctctaaag gctgtcgaaa agtctccatt tttggatgcc ttgaaggcta aaaacttcga
     4621 ggttttgttc ttgaccgacc caattgatga atacgccttc actcaattga aggaattcga
     4681 aggtaaaact ttggttgaca ttactaaaga tttcgaattg gaagaaactg acgaagaaaa
     4741 agctgaaaga gagaaggaga tcaaagaata tgaaccattg accaaagctt tgaaagaaat
     4801 tttgggtgac caagtggaga aagttgttgt ttcttacaaa ttgttggatg ccccagctgc
     4861 tatcagaact ggtcaatttg gttggtctgc taacatggaa agaatcatga aggctcaagc
     4921 cttgagagac tcttccatgt cctcctacat gtcttccaag aagactttcg aaatttctcc
     4981 aaaatctcca attatcaagg aattgaaaaa gagagttgac gaaggtggtg ctcaagacaa
     5041 gactgtcaag gacttgacta agttattata tgaaactgct ttgttgactt ccggcttcag
     5101 tttggacgaa ccaacttcct ttgcatcaag aattaacaga ttgatctctt taggcctgaa
     5161 cattgatgag gatgaagaaa cagagactgc tccagaagca tccaccgcag ctccggttga
     5221 agaggttcca gctgacaccg aaatggaaga ggtagattaa tggatcctgg tcaatccgat
     5281 gcaggtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt
     5341 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     5401 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     5461 gcgcagcctg aatggcgaat ggcgcgacgc gccctgtagc ggcgcattaa gcgcggcggg
     5521 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     5581 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     5641 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     5701 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     5761 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     5821 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     5881 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     5941 ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg cataggcaag
     6001 tgcacaaaca atacttaaat aaatactact cagtaataac ctatttctta gcatttttga
     6061 cgaaatttgc tattttgtta gagtctttta caccatttgt ctccacacct ccgcttacat
     6121 caacaccaat aacgccattt aatctaagcg catcaccaac attttctggc gtcagtccac
     6181 cagctaacat aaaatgtaag ctttcggggc tctcttgcct tccaacccag tcagaaatcg
     6241 agttccaatc caaaagttca cctgtcccac ctgcttctga atcaaacaag ggaataaacg
     6301 aatgaggttt ctgtgaagct gcactgagta gtatgttgca gtcttttgga aatacgagtc
     6361 ttttaataac tggcaaaccg aggaactctt ggtattcttg ccacgactca tctccatgca
     6421 gttggacgat atcaatgccg taatcattga ccagagccaa aacatcctcc ttaggttgat
     6481 tacgaaacac gccaaccaag tatttcggag tgcctgaact atttttatat gcttttacaa
     6541 gacttgaaat tttccttgca ataaccgggt caattgttct ctttctattg ggcacacata
     6601 taatacccag caagtcagca tcggaatcta gagcacattc tgcggcctct gtgctctgca
     6661 agccgcaaac tttcaccaat ggaccagaac tacctgtgaa attaataaca gacatactcc
     6721 aagctgcctt tgtgtgctta atcacgtata ctcacgtgct caatagtcac caatgccctc
     6781 cctcttggcc ctctcctttt cttttttcga ccgaattaat tcttaatcgg caaaaaaaga
     6841 aaagctccgg atcaagattg tacgtaaggt gacaagctat ttttcaataa agaatatctt
     6901 ccactactgc catctggcgt cataactgca aagtacacat atattacgat gctgtctatt
     6961 aaatgcttcc tatattatat atatagtaat gtcgtttatg gtgcactctc agtacaatct
     7021 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     7081 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     7141 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc ga
//
