LOCUS       sequence-232455-        8102 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8102)
  AUTHORS   Nathan DF, Lindquist S
  TITLE     Mutational analysis of Hsp90 function: interactions with a steroid 
            receptor and a protein kinase.
  JOURNAL   Mol Cell Biol 1995 Jul;15(7):3917-25.
  PUBMED    7791797
REFERENCE   2  (bases 1 to 8102)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell 
            Biol"; date: "1995-07"; volume: "15(7)"; pages: "3917-25"
COMMENT     Sequence Label: pBK2
FEATURES             Location/Qualifiers
     source          1..8102
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    55..111
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     CDS             complement(1551..2645)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(2646..3051)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     promoter        complement(4987..5005)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5026..5042)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5026..5042)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5039..5061)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5050..5066
                     /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(join(5074..5080,5081..5098,5099..5104))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5119..5140
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5257..5274)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5428..6016)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5508..5527)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6187..6978,6979..7047))
                     /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     6810..6829
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7048..7152)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7220..7238
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7276..7298)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7398..7417
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7484..7939)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7621..7642)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7833..7852
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8065..8087
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8079..8096
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8080..8096
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg cgaattgggc ccaatgcatt ggcgccgcgg ccgcgaattc
       61 gagctcggta cccggggatc ctctagagtc gacctgcagg catgcaagct taatcggatt
      121 attaactcat acgcttgtca catattgttc gaacaattct ggttctttcg agtttcgcag
      181 aactttttga atttttcttt tttttctaga acgccgtgga agaaaaacac gcgcatggtt
      241 ttatgagcgg ttaattctca tcttaatacc aaccaggtcc ttccgccacc ccctaaaaca
      301 tataaatatg cagcttatcc cttcaattct taacatctgt gacctcctca tttcttcccg
      361 ctgtattaga gttcaagaaa tcatacctga tagaaaatag agtcctataa acaaaagcac
      421 aaacaaacac gcaaagatat ggctagtgaa acttttgaat ttcaagctga aattactcag
      481 ttgatgagtt tgatcatcaa caccgtctat tctaacaagg aaattttctt gagagaactg
      541 atatctaatg cctcggatgc gttggataaa attagataca aatctttgtc tgatccaaag
      601 caattggaaa cagaaccaga tctctttatt agaatcactc caaagccaga gcaaaaagtt
      661 ttggaaatca gagattctgg tattggtatg accaaggatc tggtactttg gcccccccat
      721 tggtagcggg atttggtgaa gctgcgagat tgatgaagaa agaatttgac aacgaccaag
      781 ctcacatcaa aagactatcc gataaattag tcaaaggtct attatccgct gaacatacca
      841 cgttgaacgg atctccagat catcgttatc cagggtgtgt taacgtttct ttcgcctacg
      901 tggaaggaga atctttattg atggcactaa gggatatcgc attatcctcg ggttcagcct
      961 gtacatctgc ttccctagaa ccttcttatg ttttacatgc gctgggtaag gatgatgcat
     1021 tagcccattc ttccatcaga tttggtattg gtagatttag tactgaagag gaggtcgact
     1081 acgtcgttaa ggccgtttct gacagagtaa aattcttgag ggaactttca ccattatggg
     1141 aaatggttca agaaggtatt gacttaaact ccatcaaatg gtcaggtcat tgagtgtttt
     1201 ttatttgttg tatttttttt ttttagagaa aatcctccaa tatataaatt aggaatcata
     1261 gtttcatgat tttctgttac acctaacttt ttgtgtggtg ccctcctcct tgtcaatatt
     1321 aatgttaaag tgcaattctt tttccttatc acgttgagcc attagtatca atttgcttac
     1381 ctgtattcct ttacatcctc ctttttctcc ttcttgataa atgtatgtag attgcgtata
     1441 tagtttcgtc taccctatga acatattcca ttttgtaatt tcgtgtcgtt tctattatga
     1501 atttcattta taaagtttat gtacaaatat cataaaaaaa gagaatcttt ttaagcaagg
     1561 attttcttaa cttcttcggc gacagcatca ccgacttcgg tggtactgtt ggaaccacct
     1621 aaatcaccag ttctgatacc tgcatccaaa acctttttaa ctgcatcttc aatggcctta
     1681 ccttcttcag gcaagttcaa tgacaatttc aacatcattg cagcagacaa gatagtggcg
     1741 atagggttga ccttattctt tggcaaatct ggagcagaac cgtggcatgg ttcgtacaaa
     1801 ccaaatgcgg tgttcttgtc tggcaaagag gccaaggacg cagatggcaa caaacccaag
     1861 gaacctggga taacggaggc ttcatcggag atgatatcac caaacatgtt gctggtgatt
     1921 ataataccat ttaggtgggt tgggttctta actaggatca tggcggcaga atcaatcaat
     1981 tgatgttgaa ccttcaatgt agggaattcg ttcttgatgg tttcctccac agtttttctc
     2041 cataatcttg aagaggccaa aacattagct ttatccaagg accaaatagg caatggtggc
     2101 tcatgttgta gggccatgaa agcggccatt cttgtgattc tttgcacttc tggaacggtg
     2161 tattgttcac tatcccaagc gacaccatca ccatcgtctt cctttctctt accaaagtaa
     2221 atacctccca ctaattctct gacaacaacg aagtcagtac ctttagcaaa ttgtggcttg
     2281 attggagata agtctaaaag agagtcggat gcaaagttac atggtcttaa gttggcgtac
     2341 aattgaagtt ctttacggat ttttagtaaa ccttgttcag gtctaacact accggtaccc
     2401 catttaggac cacccacagc acctaacaaa acggcatcag ccttcttgga ggcttccagc
     2461 gcctcatctg gaagtggaac acctgtagca tcgatagcag caccaccaat taaatgattt
     2521 tcgaaatcga acttgacatt ggaacgaaca tcagaaatag ctttaagaac cttaatggct
     2581 tcggctgtga tttcttgacc aacgtggtca cctggcaaaa cgacgatctt cttaggggca
     2641 gacattagaa tggtatatcc ttgaaatata tatatatatt gctgaaatgt aaaaggtaag
     2701 aaaagttaga aagtaagacg attgctaacc acctattgga aaaaacaata ggtccttaaa
     2761 taatattgtc aacttcaagt attgtgatgc aagcatttag tcatgaacgc ttctctattc
     2821 tatatgaaaa gccggttccg gcgctctcac ctttcctttt tctcccaatt tttcagttga
     2881 aaaaggtata tgcgtcaggc gacctctgaa attaacaaaa aatttccagt catcgaattt
     2941 gattctgtgc gatagcgccc ctgtgtgttc tcgttatgtt gaggaaaaaa ataatggttg
     3001 ctaagagatt cgaactcttg catcttacga tacctgagta ttcccacagt taactgcggt
     3061 caagatattt cttgaatcag gcgccttaga ccgctcggcc aaacaaccaa ttacttgttg
     3121 agaaatagag tataattatc ctataaatat aacgtttttg aacacacatg aacaaggaag
     3181 tacaggacaa ttgattttga agagaatgtg gattttgatg taattgttgg gattccattt
     3241 ttaataaggc aataatatta ggtatgtaga tatactagaa gttctcctcg aggatttagg
     3301 aatccataaa agggaatctg caattctaca caattctata aatattatta tcatcatttt
     3361 atatgttaat attcattgat cctattacat tatcaatcct tgcgtttcag cttccactaa
     3421 tttagatgac tatttctcat catttgcgtc atcttctaac accgtatatg ataatatact
     3481 agtaacgtaa atactagtta gtagatgata gttgattttt attccaacat accacccata
     3541 atgtaataga tccaaggcct tgaaagaaat tttgggtgac caagtggaga aagttgttgt
     3601 ttcttacaaa ttgttggatg ccccagctgc tatcagaact ggtcaatttg gttggtctgc
     3661 taacatggaa agaatcatga aggctcaagc cttgagagac tcttccatgt cctcctacat
     3721 gtcttccaag aagactttcg aaatttctcc aaaatctcca attatcaagg aattgaaaaa
     3781 gagagttgac gaaggtggtg ctcaagacaa gactgtcaag gacttgacta agttattata
     3841 tgaaactgct ttgttgactt ccggcttcag tttggacgaa ccaacttcct ttgcatcaag
     3901 aattaacaga ttgatctctt tgggtttgaa cattgatgag gatgaagaaa cagagactgc
     3961 tccagaagca tccaccgcag ctccggttga agaggttcca gctgacaccg aaatggaaga
     4021 ggtagattag gtagaacatc atggccttga ataggttata aacaaaacat aatataacgt
     4081 ataggtattc gaatgaataa ataagtatgt aaatagggca tctgcatgga aataactggg
     4141 taaaacatta caatgtattt ttttagagtt ttcactattc cgacgcgtct agtgtgtcat
     4201 acaaaacgcg aagaatctga aatcgtgaaa tatacttaag aatataactg aaatagctta
     4261 agcataaatg gactttactg cgaagataaa gggtatgttg aaactcatct gctttcatta
     4321 tgcgtccaac taattaacat tgcatatttt cgtatactat ttttttgtgt agagctcgaa
     4381 cgagaactct cagagacgag tgactataag accttgcaga agaagaccat aagtcttcga
     4441 agtgagctga atactctgtc acattctttg acatcttatg aaaaagaaca cttctccaat
     4501 gatatagaaa atgtattaaa atctattaat gcaaaattat ctgagtcaaa gggcaaaaaa
     4561 cgactattct ccttcaaaca gaaaaattca tcatcagccg ttcataagaa tgttgaacgt
     4621 acggagcttg cgaatgcgcc ggcgtatacc actactttga aaaagcatta tgtcttggag
     4681 aagggtgatt cagcttttga aaatttagag ttttgtactg ttacatcaac aacagattat
     4741 agtggcaata gtgccctttc tggttctttg tgctttcgca atattactaa atgtgtcata
     4801 aacttacaaa ggattttctt tcaaacaggg agtattttca taactgattg tacggattct
     4861 ataatttttc tacggtctcc ttccgacaag gatttccaaa tccgtttacg tgacttgaaa
     4921 aattgcaaaa ttttgataga gaaactctcg ccaagtatcg acctgcaggc atgcaagctt
     4981 ttgttccctt tagtgagggt taatttcgag cttggcgtaa tcatggtcat agctgtttcc
     5041 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5101 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5161 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5221 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5281 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5341 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5401 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5461 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5521 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5581 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5641 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5701 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5761 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5821 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     5881 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5941 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6001 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6061 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6121 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6181 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6241 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6301 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6361 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6421 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6481 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6541 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6601 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6661 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6721 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6781 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6841 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6901 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6961 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7021 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7081 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7141 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     7201 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     7261 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     7321 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     7381 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     7441 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggaaattgt aagcgttaat
     7501 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     7561 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     7621 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     7681 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     7741 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     7801 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     7861 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     7921 gcgccgctac agggcgcgtc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     7981 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     8041 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat
     8101 tg
//
