LOCUS       sequence-237099-        8456 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 8456)
  AUTHORS   Schirmer EC, Homann OR, Kowal AS, Lindquist S
  TITLE     Dominant gain-of-function mutations in Hsp104p reveal crucial roles 
            for the middle region.
  JOURNAL   Mol Biol Cell 2004 May;15(5):2061-72. Epub 2004 Feb 20.
  PUBMED    14978213
REFERENCE   2  (bases 1 to 8456)
  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 Biol 
            Cell"; date: "2004-05"; volume: "15(5)"; pages: "2061-72. Epub 2004 
            Feb 20"
COMMENT     Sequence Label: pGALSc104(T499I)
FEATURES             Location/Qualifiers
     source          1..8456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     promoter        317..504
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             505..1167
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     rep_origin      complement(1427..1882)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1564..1585)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1776..1795
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2008..2030
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2022..2039
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2023..2039
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2046..2065
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2046..2064
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4935..5376)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(4941..4964)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    5259..5376
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     promoter        complement(5683..5701)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5722..5738)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5722..5738)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5735..5757)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5746..5762
                     /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(5770..5776,5777..5794,5795..5800))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5815..5836
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5953..5970)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6124..6712)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6204..6223)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6883..7674,7675..7743))
                     /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     7506..7525
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7744..7848)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7878..8381
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     8429..8447
                     /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 accataaatt cccgttttaa gagcttggtg agcgctagga gtcactgcca ggtatcgttt
      241 gaacacggca ttagtcaggg aagtcataac acagtccttt cccgcaattt tctttttcta
      301 ttactcttgg cctcctctag tacactctat atttttttat gcctcggtaa tgattttcat
      361 tttttttttt ccacctagcg gatgactctt tttttttctt agcgattggc attatcacat
      421 aatgaattat acattatata aagtaatgtg atttcttcga agaatatact aaaaaatgag
      481 caggcaagat aaacgaaggc aaagatgaca gagcagaaag ccctagtaaa gcgtattaca
      541 aatgaaacca agattcagat tgcgatctct ttaaagggtg gtcccctagc gatagagcac
      601 tcgatcttcc cagaaaaaga ggcagaagca gtagcagaac aggccacaca atcgcaagtg
      661 attaacgtcc acacaggtat agggtttctg gaccatatga tacatgctct ggccaagcat
      721 tccggctggt cgctaatcgt tgagtgcatt ggtgacttac acatagacga ccatcacacc
      781 actgaagact gcgggattgc tctcggtcaa gcttttaaag aggccctagg ggccgtgcgt
      841 ggagtaaaaa ggtttggatc aggatttgcg cctttggatg aggcactttc cagagcggtg
      901 gtagatcttt cgaacaggcc gtacgcagtt gtcgaacttg gtttgcaaag ggagaaagta
      961 ggagatctct cttgcgagat gatcccgcat tttcttgaaa gctttgcaga ggctagcaga
     1021 attaccctcc acgttgattg tctgcgaggc aagaatgatc atcaccgtag tgagagtgcg
     1081 ttcaaggctc ttgcggttgc cataagagaa gccacctcgc ccaatggtac caacgatgtt
     1141 ccctccacca aaggtgttct tatgtagtga caccgattat ttaaagctgc agcatacgat
     1201 atatatacat gtgtatatat gtatacctat gaatgtcagt aagtatgtat acgaacagta
     1261 tgatactgaa gatgacaagg taatgcatca ttctatacgt gtcattctga acgaggcgcg
     1321 ctttcctttt ttctttttgc tttttctttt tttttctctt gaactcgacg gatctatgcg
     1381 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat tgtaagcgtt
     1441 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     1501 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     1561 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     1621 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     1681 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     1741 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     1801 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     1861 aatgcgccgc tacagggcgc gtccattcgc cattcaggct gcgcaactgt tgggaagggc
     1921 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     1981 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     2041 aattgtaata cgactcacta tagggcgaat tggagctctt ttgctcgggt gtcaagttca
     2101 attaaacaaa ttccattagt tttattaatt atatatatta tattactgat tcttgttcga
     2161 aagtttttaa aaatcacact atattaaatt aatctaggtc atcatcaatt tccatactgt
     2221 cctcattatc gtcatcacct aacgtgtcag cccctatagt agcttcgtga tttggtagaa
     2281 cttccagaca ttcttcagct tcctcaggaa cattttcatc acgagattta cccttcttca
     2341 agacgacatt gacagtttcc ttatccttga tttcattctt taagatcctt agtgccagtt
     2401 tgttcaaaat ttcgttttga attaacctgt tcagtggacg tgcacccata tcatcggaat
     2461 aaccatattt ggccaagaag tccttggcct cttgagttaa attcaacttg taatgtttat
     2521 cattttgctc gaatctctct tcaatttcct tcaaacgaat atccacgatc ttatgaatag
     2581 cttttctaga tagcttgttg aaaatgacta tactagaaat tctgttcaaa aattctggtc
     2641 tgaaatgttg cctaacagca cccatgacca aattcttggt agattcttgg atctttgatc
     2701 cttgttgaga attgataaat tcagcaccta gattggaagt catgatgaca atacaattgg
     2761 aacagtcgat cgtcttacct tgaccactag taattctacc gtcatccaac atttgtagca
     2821 tgacagtcaa aacatcagga tgtgcctttt ctacttcatc gaataacaaa acggagtatg
     2881 gtttgtattg cagttggtta gttaaaaagc caccttcatc gtacccgaca taacctgccg
     2941 tggtacccaa caacttagag accgcatact tctcgcttaa ttcagaacaa tcgaccctga
     3001 tcatcatgtc ctcatcatta aacaaaaatc cagcaacttt tttagccaat tcagttttac
     3061 cggaaccgga caaacctaaa aataagaagg atgctggttg ccttggatta gctaaacctg
     3121 atctagacaa tctaacggca ttggaaacag ctttaatggc atccatttgg cccacgactt
     3181 cagatgataa gtcacgttcc atatgaatca atttttcatt ttcagattct gacaacttct
     3241 taacagggat accagtcaat cttgcagctg tttcagaaat ggtgtctgag tcgaccacat
     3301 tttggatcat ggagttggca ccagcacgtc tctcttcctc agcaacctga tcttcaagct
     3361 tttcgatttg ctttttgata tctgggatgg cgaagtacct taaatcagcg gcggtagcaa
     3421 tatcatatct acgttcagca tcaagggcct tgttttccag ttcatccaat ttctttttag
     3481 cttgtgtcaa ttcttcatgg ccatgctttt cttcattgta acgttgtctt agaggttcca
     3541 attcttcttg caatgaagct tccttctgcc tagctaactt taatctatct ttagtggtgg
     3601 agtcggcatc ttcatctctc tctagagctt ttatctctac ttgaatcaat tgcaattgac
     3661 gttccttgga atccaattct tctggcttag aatctcttgc gacggcgaca ccagcacaag
     3721 aaatatcaac taaatccaaa gcagaatctg gcaatcttct atatggcaag taacgcttgg
     3781 ctaattgagc cgcggtgact aaggcgctat ccagaatcct tacaccatga tgtatttcat
     3841 actttggttg cagacctctc aatatggcca ctgtttgtct cacacttggt tcagcgactt
     3901 caattttctg gaatcttctt tcaaaggcac catccttttc cacaatagat ctatattcgt
     3961 tattggtggt ggcaccgatg accttcaatt ggcctctgga caaagctggc ttcaagatgt
     4021 tagcagcgtc gtccttacca ttacccatta acatgtgaat ttcatcaatg aataacacaa
     4081 ttagagtctt tgattcttcg atttccttca aaacaccttt gaatctttct tcgaaatcac
     4141 ctttgtattt agcacctgcg gttaatgcgg ccaaatctag actgaacaat ttagcgcctt
     4201 gtaagatagt gggaacgtca tcgtcaatga ttctttgagc aacaccttca ataatagcgg
     4261 tcttaccgat acctggctca ccaattaaac atgggttgga cttaattctt cttgctaaaa
     4321 ctctaatagt gcttcttatt tcttcttcac ggccgatgac agggtcaagt ttaccttgac
     4381 gagcctgctc agtcatatca atggcgtact ttgataaata ttccaaaggt gtgttcgtat
     4441 cagcgccacg agagtcaatt ctagtgttac cacgaagttc aagagcttgt tgcttgatgg
     4501 cctcaatatc tacttgagct tccttaaata tttgctgaat agacgaatca ttgaatagag
     4561 caaacaatat atggtcttgc gctataaatg agtccttctg ttgtttttga atcttagcag
     4621 cgtcttgaag gactttcccc aaagcataac ttggagttat ctccgcaggt gcaggttgct
     4681 gttgaggaat tcttactaga tttctattaa ccactttctt gaaaagatca tagtcgtaac
     4741 ggcccttctc aattagattc tgtaggtaag ggactgatcc atcttctggc gtttcaatga
     4801 aggcagctag aatatgtata ggttgtaatt gtggatgttg atgatccgaa gccaattttt
     4861 gagccaacgt caaaatcgtt agagcccttt ctgtaaattg cgtttggtcg ttcatcccgg
     4921 atccggggtt ttttctcctt gacgttaaag tatagaggta tattaacaat tttttgttga
     4981 tacttttatt acatttgaat aagaagtaat acaaaccgaa aatgttgaaa gtattagtta
     5041 aagtggttat gcagtttttg catttatata tctgttaata gatcaaaaat catcgcttcg
     5101 ctgattaatt accccagaaa taaggctaaa aaactaatcg cattatcatc ctatggttgt
     5161 taatttgatt cgttcatttg aaggtttgtg gggccaggtt actgccaatt tttcctcttc
     5221 ataaccataa aagctagtat tgtagaatct ttattgttcg gagcagtgcg gcgcgaggca
     5281 catctgcgtt tcaggaacgc gaccggtgaa gacgaggacg cacggaggag agtcttcctt
     5341 cggagggctg tcacccgctc ggcggcttct aatccgtact tcaatatagc aatgagcagt
     5401 taagcgtatt actgaaagtt ccaaagagaa ggttttttta ggctaagata atggggctct
     5461 ttacatttcc acaacatata agtaagatta gatatggata tgtatatgga tatgtatatg
     5521 gatatgtata tggatatgta tatggtggta atgccatgta atatgattat taaacttctt
     5581 tgcgtccatc caaaaaaaaa gtaagaattt ttgaaaattc aaggaattcg atatcaagct
     5641 tatcgatacc gtcgaggggg ggcccggtac cagcttttgt tccctttagt gagggttaat
     5701 ttcgagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5761 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5821 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5881 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5941 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6001 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6061 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6121 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6181 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6241 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6301 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6361 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6421 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6481 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6541 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6601 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6661 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6721 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6781 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6841 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6901 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6961 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7021 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7081 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7141 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7201 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7261 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7321 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7381 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7441 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7501 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7561 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7621 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7681 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7741 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7801 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7861 aaaagtgcca cctgacggat cgcttgcctg taacttacac gcgcctcgta tcttttaatg
     7921 atggaataat ttgggaattt actctgtgtt tatttatttt tatgttttgt atttggattt
     7981 tagaaagtaa ataaagaagg tagaagagtt acggaatgaa gaaaaaaaaa taaacaaagg
     8041 tttaaaaaat ttcaacaaaa agcgtacttt acatatatat ttattagaca agaaaagcag
     8101 attaaataga tatacattcg attaacgata agtaaaatgt aaaatcacag gattttcgtg
     8161 tgtggtcttc tacacagaca agatgaaaca attcggcatt aatacctgag agcaggaaga
     8221 gcaagataaa aggtagtatt tgttggcgat ccccctagag tcttttacat cttcggaaaa
     8281 caaaaactat tttttcttta atttcttttt ttactttcta tttttaattt atatatttat
     8341 attaaaaaat ttaaattata attattttta tagcacgtga tgaaaaggac cctaagaaac
     8401 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc
//
