LOCUS       sequence-238258-        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(G217S/T499I)
FEATURES             Location/Qualifiers
     source          1..8456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    55..76
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(91..96,97..114,115..121)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    129..145
                     /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     134..156
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     153..169
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     153..169
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        190..208
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        515..956
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    515..632
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     927..950
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(3826..3845)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3827..3845)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3852..3869)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3852..3868)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3861..3883)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4009..4464
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4096..4115)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4306..4327
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4724..5386)
                     /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"
     promoter        complement(5387..5574)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(5721..5740)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5840..5862
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5900..5918)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5966..6469
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        6499..6603
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6604..6672,6673..7464)
                     /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(6822..6841)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7635..8223
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8124..8143
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8377..8394
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
       61 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
      121 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
      181 caagctcgaa attaaccctc actaaaggga acaaaagctg gtaccgggcc ccccctcgac
      241 ggtatcgata agcttgatat cgaattcctt gaattttcaa aaattcttac tttttttttg
      301 gatggacgca aagaagttta ataatcatat tacatggcat taccaccata tacatatcca
      361 tatacatatc catatacata tccatataca tatccatatc taatcttact tatatgttgt
      421 ggaaatgtaa agagccccat tatcttagcc taaaaaaacc ttctctttgg aactttcagt
      481 aatacgctta actgctcatt gctatattga agtacggatt agaagccgcc gagcgggtga
      541 cagccctccg aaggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga
      601 aacgcagatg tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt
      661 ttatggttat gaagaggaaa aattggcagt aacctggccc cacaaacctt caaatgaacg
      721 aatcaaatta acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt
      781 aattaatcag cgaagcgatg atttttgatc tattaacaga tatataaatg caaaaactgc
      841 ataaccactt taactaatac tttcaacatt ttcggtttgt attacttctt attcaaatgt
      901 aataaaagta tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa
      961 aaccccggat ccgggatgaa cgaccaaacg caatttacag aaagggctct aacgattttg
     1021 acgttggctc aaaaattggc ttcggatcat caacatccac aattacaacc tatacatatt
     1081 ctagctgcct tcattgaaac gccagaagat ggatcagtcc cttacctaca gaatctaatt
     1141 gagaagggcc gttacgacta tgatcttttc aagaaagtgg ttaatagaaa tctagtaaga
     1201 attcctcaac agcaacctgc acctgcggag ataactccaa gttatgcttt ggggaaagtc
     1261 cttcaagacg ctgctaagat tcaaaaacaa cagaaggact catttatagc gcaagaccat
     1321 atattgtttg ctctattcaa tgattcgtct attcagcaaa tatttaagga agctcaagta
     1381 gatattgagg ccatcaagca acaagctctt gaacttcgtg gtaacactag aattgactct
     1441 cgtggcgctg atacgaacac acctttggaa tatttatcaa agtacgccat tgatatgact
     1501 gagcaggctc gtcaaggtaa acttgaccct gtcatcggcc gtgaagaaga aataagaagc
     1561 actattagag ttttagcaag aagaattaag tccaacccat gtttaattgg tgagccaggt
     1621 atcagtaaga ccgctattat tgaaggtgtt gctcaaagaa tcattgacga tgacgttccc
     1681 actatcttac aaggcgctaa attgttcagt ctagatttgg ccgcattaac cgcaggtgct
     1741 aaatacaaag gtgatttcga agaaagattc aaaggtgttt tgaaggaaat cgaagaatca
     1801 aagactctaa ttgtgttatt cattgatgaa attcacatgt taatgggtaa tggtaaggac
     1861 gacgctgcta acatcttgaa gccagctttg tccagaggcc aattgaaggt catcggtgcc
     1921 accaccaata acgaatatag atctattgtg gaaaaggatg gtgcctttga aagaagattc
     1981 cagaaaattg aagtcgctga accaagtgtg agacaaacag tggccatatt gagaggtctg
     2041 caaccaaagt atgaaataca tcatggtgta aggattctgg atagcgcctt agtcaccgcg
     2101 gctcaattag ccaagcgtta cttgccatat agaagattgc cagattctgc tttggattta
     2161 gttgatattt cttgtgctgg tgtcgccgtc gcaagagatt ctaagccaga agaattggat
     2221 tccaaggaac gtcaattgca attgattcaa gtagagataa aagctctaga gagagatgaa
     2281 gatgccgact ccaccactaa agatagatta aagttagcta ggcagaagga agcttcattg
     2341 caagaagaat tggaacctct aagacaacgt tacaatgaag aaaagcatgg ccatgaagaa
     2401 ttgacacaag ctaaaaagaa attggatgaa ctggaaaaca aggcccttga tgctgaacgt
     2461 agatatgata ttgctaccgc cgctgattta aggtacttcg ccatcccaga tatcaaaaag
     2521 caaatcgaaa agcttgaaga tcaggttgct gaggaagaga gacgtgctgg tgccaactcc
     2581 atgatccaaa atgtggtcga ctcagacacc atttctgaaa cagctgcaag attgactggt
     2641 atccctgtta agaagttgtc agaatctgaa aatgaaaaat tgattcatat ggaacgtgac
     2701 ttatcatctg aagtcgtggg ccaaatggat gccattaaag ctgtttccaa tgccgttaga
     2761 ttgtctagat caggtttagc taatccaagg caaccagcat ccttcttatt tttaggtttg
     2821 tccggttccg gtaaaactga attggctaaa aaagttgctg gatttttgtt taatgatgag
     2881 gacatgatga tcagggtcga ttgttctgaa ttaagcgaga agtatgcggt ctctaagttg
     2941 ttgggtacca cggcaggtta tgtcgggtac gatgaaggtg gctttttaac taaccaactg
     3001 caatacaaac catactccgt tttgttattc gatgaagtag aaaaggcaca tcctgatgtt
     3061 ttgactgtca tgctacaaat gttggatgac ggtagaatta ctagtggtca aggtaagacg
     3121 atcgactgtt ccaattgtat tgtcatcatg acttccaatc taggtgctga atttatcaat
     3181 tctcaacaag gatcaaagat ccaagaatct accaagaatt tggtcatggg tgctgttagg
     3241 caacatttca gaccagaatt tttgaacaga atttctagta tagtcatttt caacaagcta
     3301 tctagaaaag ctattcataa gatcgtggat attcgtttga aggaaattga agagagattc
     3361 gagcaaaatg ataaacatta caagttgaat ttaactcaag aggccaagga cttcttggcc
     3421 aaatatggtt attccgatga tatgggtgca cgtccactga acaggttaat tcaaaacgaa
     3481 attttgaaca aactggcact aaggatctta aagaatgaaa tcaaggataa ggaaactgtc
     3541 aatgtcgtct tgaagaaggg taaatctcgt gatgaaaatg ttcctgagga agctgaagaa
     3601 tgtctggaag ttctaccaaa tcacgaagct actatagggg ctgacacgtt aggtgatgac
     3661 gataatgagg acagtatgga aattgatgat gacctagatt aatttaatat agtgtgattt
     3721 ttaaaaactt tcgaacaaga atcagtaata taatatatat aattaataaa actaatggaa
     3781 tttgtttaat tgaacttgac acccgagcaa aagagctcca attcgcccta tagtgagtcg
     3841 tattacaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     3901 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     3961 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggac gcgccctgta
     4021 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4081 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4141 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4201 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     4261 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4321 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     4381 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     4441 acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg
     4501 tatttcacac cgcatagatc cgtcgagttc aagagaaaaa aaaagaaaaa gcaaaaagaa
     4561 aaaaggaaag cgcgcctcgt tcagaatgac acgtatagaa tgatgcatta ccttgtcatc
     4621 ttcagtatca tactgttcgt atacatactt actgacattc ataggtatac atatatacac
     4681 atgtatatat atcgtatgct gcagctttaa ataatcggtg tcactacata agaacacctt
     4741 tggtggaggg aacatcgttg gtaccattgg gcgaggtggc ttctcttatg gcaaccgcaa
     4801 gagccttgaa cgcactctca ctacggtgat gatcattctt gcctcgcaga caatcaacgt
     4861 ggagggtaat tctgctagcc tctgcaaagc tttcaagaaa atgcgggatc atctcgcaag
     4921 agagatctcc tactttctcc ctttgcaaac caagttcgac aactgcgtac ggcctgttcg
     4981 aaagatctac caccgctctg gaaagtgcct catccaaagg cgcaaatcct gatccaaacc
     5041 tttttactcc acgcacggcc cctagggcct ctttaaaagc ttgaccgaga gcaatcccgc
     5101 agtcttcagt ggtgtgatgg tcgtctatgt gtaagtcacc aatgcactca acgattagcg
     5161 accagccgga atgcttggcc agagcatgta tcatatggtc cagaaaccct atacctgtgt
     5221 ggacgttaat cacttgcgat tgtgtggcct gttctgctac tgcttctgcc tctttttctg
     5281 ggaagatcga gtgctctatc gctaggggac caccctttaa agagatcgca atctgaatct
     5341 tggtttcatt tgtaatacgc tttactaggg ctttctgctc tgtcatcttt gccttcgttt
     5401 atcttgcctg ctcatttttt agtatattct tcgaagaaat cacattactt tatataatgt
     5461 ataattcatt atgtgataat gccaatcgct aagaaaaaaa aagagtcatc cgctaggtgg
     5521 aaaaaaaaaa atgaaaatca ttaccgaggc ataaaaaaat atagagtgta ctagaggagg
     5581 ccaagagtaa tagaaaaaga aaattgcggg aaaggactgt gttatgactt ccctgactaa
     5641 tgccgtgttc aaacgatacc tggcagtgac tcctagcgct caccaagctc ttaaaacggg
     5701 aatttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     5761 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     5821 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     5881 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     5941 ataatggttt cttagggtcc ttttcatcac gtgctataaa aataattata atttaaattt
     6001 tttaatataa atatataaat taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag
     6061 tttttgtttt ccgaagatgt aaaagactct agggggatcg ccaacaaata ctacctttta
     6121 tcttgctctt cctgctctca ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag
     6181 accacacacg aaaatcctgt gattttacat tttacttatc gttaatcgaa tgtatatcta
     6241 tttaatctgc ttttcttgtc taataaatat atatgtaaag tacgcttttt gttgaaattt
     6301 tttaaacctt tgtttatttt tttttcttca ttccgtaact cttctacctt ctttatttac
     6361 tttctaaaat ccaaatacaa aacataaaaa taaataaaca cagagtaaat tcccaaatta
     6421 ttccatcatt aaaagatacg aggcgcgtgt aagttacagg caagcgatcc gtcaggtggc
     6481 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     6541 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     6601 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     6661 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     6721 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     6781 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     6841 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     6901 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     6961 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     7021 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     7081 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     7141 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     7201 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     7261 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     7321 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     7381 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     7441 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     7501 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     7561 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     7621 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     7681 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     7741 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     7801 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     7861 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     7921 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     7981 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     8041 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     8101 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     8161 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     8221 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     8281 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     8341 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     8401 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatg
//
