LOCUS       sequence-197910-        6851 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 6851)
  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 6851)
  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: GAL-CDC12(E368Q)
FEATURES             Location/Qualifiers
     source          1..6851
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    22..43
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(160..177)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(331..919)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(411..430)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1090..1881,1882..1950))
                     /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     1713..1732
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1951..2055)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    2092..2595
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     2636..2654
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2692..2714)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2814..2833
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2857..3072
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             3073..3876
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(4007..4462)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4144..4165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4356..4375
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4588..4610
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4602..4619
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4603..4619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4626..4645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4626..4644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        6004..6668
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     primer_bind     complement(6017..6040)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    6335..6452
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     complement(6696..6712)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6697..6713)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(6741..6759)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6780..6796)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6780..6796)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6793..6815)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6804..6820
                     /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(6828..6834,6835..6851,1,2..7))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
       61 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
      121 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
      181 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
      241 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
      301 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
      361 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
      421 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
      481 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
      541 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
      601 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
      661 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
      721 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
      781 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
      841 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
      901 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
      961 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     1021 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     1081 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     1141 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     1201 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     1261 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     1321 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     1381 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     1441 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     1501 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     1561 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     1621 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     1681 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     1741 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     1801 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     1861 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     1921 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     1981 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     2041 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gggtcctttt catcacgtgc
     2101 tataaaaata attataattt aaatttttta atataaatat ataaattaaa aatagaaagt
     2161 aaaaaaagaa attaaagaaa aaatagtttt tgttttccga agatgtaaaa gactctaggg
     2221 ggatcgccaa caaatactac cttttatctt gctcttcctg ctctcaggta ttaatgccga
     2281 attgtttcat cttgtctgtg tagaagacca cacacgaaaa tcctgtgatt ttacatttta
     2341 cttatcgtta atcgaatgta tatctattta atctgctttt cttgtctaat aaatatatat
     2401 gtaaagtacg ctttttgttg aaatttttta aacctttgtt tatttttttt tcttcattcc
     2461 gtaactcttc taccttcttt atttactttc taaaatccaa atacaaaaca taaaaataaa
     2521 taaacacaga gtaaattccc aaattattcc atcattaaaa gatacgaggc gcgtgtaagt
     2581 tacaggcaag cgatccgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     2641 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     2701 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     2761 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     2821 agagcagatt gtactgagag tgcaccatac caccttttca attcatcatt ttttttttat
     2881 tctttttttt gatttcggtt tccttgaaat ttttttgatt cggtaatctc cgaacagaag
     2941 gaagaacgaa ggaaggagca cagacttaga ttggtatata tacgcatatg tagtgttgaa
     3001 gaaacatgaa attgcccagt attcttaacc caactgcaca gaacaaaaac ctgcaggaaa
     3061 cgaagataaa tcatgtcgaa agctacatat aaggaacgtg ctgctactca tcctagtcct
     3121 gttgctgcca agctatttaa tatcatgcac gaaaagcaaa caaacttgtg tgcttcattg
     3181 gatgttcgta ccaccaagga attactggag ttagttgaag cattaggtcc caaaatttgt
     3241 ttactaaaaa cacatgtgga tatcttgact gatttttcca tggagggcac agttaagccg
     3301 ctaaaggcat tatccgccaa gtacaatttt ttactcttcg aagacagaaa atttgctgac
     3361 attggtaata cagtcaaatt gcagtactct gcgggtgtat acagaatagc agaatgggca
     3421 gacattacga atgcacacgg tgtggtgggc ccaggtattg ttagcggttt gaagcaggcg
     3481 gcagaagaag taacaaagga acctagaggc cttttgatgt tagcagaatt gtcatgcaag
     3541 ggctccctat ctactggaga atatactaag ggtactgttg acattgcgaa gagcgacaaa
     3601 gattttgtta tcggctttat tgctcaaaga gacatgggtg gaagagatga aggttacgat
     3661 tggttgatta tgacacccgg tgtgggttta gatgacaagg gagacgcatt gggtcaacag
     3721 tatagaaccg tggatgatgt ggtctctaca ggatctgaca ttattattgt tggaagagga
     3781 ctatttgcaa agggaaggga tgctaaggta gagggtgaac gttacagaaa agcaggctgg
     3841 gaagcatatt tgagaagatg cggccagcaa aactaaaaaa ctgtattata agtaaatgca
     3901 tgtatactaa actcacaaat tagagcttca atttaattat atcagttatt accctatgcg
     3961 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat tgtaagcgtt
     4021 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     4081 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     4141 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     4201 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     4261 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     4321 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     4381 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     4441 aatgcgccgc tacagggcgc gtccattcgc cattcaggct gcgcaactgt tgggaagggc
     4501 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     4561 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     4621 aattgtaata cgactcacta tagggcgaat tggagctcca ccgcggtggc ggccgctcta
     4681 gatatgtgcg tattgctgcg acaaaagaag agacaaagag ataggcgttg aaattgacga
     4741 gacaaagagg aagacattaa ttaatcatca ttttaaatgg gattttttta cttgcaagct
     4801 tttgacctgc tcttctagtt tttttacctt tccttgaatt tcttccaagt caccgttcag
     4861 tctgattctt tcgttaacaa tgttttgctg ccattgtcta aatctttgct cctcagcttt
     4921 gacttggtca gtgaaatact tcttcaatgc attttcttct tctttgtatc taggattgtg
     4981 tgacaatttc cttgctggtg caattgctgg aagagtgaaa tcttcattcc cctccccggt
     5041 gttctcgtga ccctcaagtc ttaatcttct gtatgtttcg taatgcattt cttgggtggt
     5101 agaaatcaaa tctaatagat aagtcctcag taacagggct ctgagcttac ggaaatcaca
     5161 atgagaatcg ttttcgattt ccacaagtcc ccaaggatac ttccttgcaa ctacctgagt
     5221 gccttgccca ttgtcaaatt ttttttcaga ccctactatg gcaaatggca ttgcttctat
     5281 caactgtctt gcatgttcaa cagcggcaga atctggatta ctgccacttt tggcatcttc
     5341 cttagaatca gcgtctaatg gaggcgtgaa aatacgaatt tcttgagctt ctattacttg
     5401 cctgattctg gacttgaact gttgtaattc ttgggcagtc aatgtatcag ctttcgcaat
     5461 aactggaatc agattggctc tggttgacaa tctcttcata gtctcaatgt caattggctt
     5521 taaaccatgt ccagtgggcc tgatgaagta cagaacggca tgaactctca agtcaaattt
     5581 cttagtacgg tacggttgct gctcttgacg catataagaa tcgtgttgat catcaataaa
     5641 atccaccaat ggttgccagg ctttgttgtt attcacgtta tctccaaaac ctggggtgtc
     5701 gatgacatta acgcgtaatt cgaaatgctt ctcttccagc aaagctcttg tgatgtcgat
     5761 ttctacagtc tttctgatgg gttcttggcg gtgttgctga ccatctgccc ttttcaagac
     5821 agtctgaaat aaggtattaa taaatgtagt tttacccaat ccgctctcac cacataacat
     5881 cacggtgaat gtaccacctt cttcattaac aattttataa cgctggttgg gaagattgga
     5941 tatccctaca ggaggaggaa caggggcagc agtcgcagtg gcagcactca ttatggatcc
     6001 ggggtttttt ctccttgacg ttaaagtata gaggtatatt aacaattttt tgttgatact
     6061 tttattacat ttgaataaga agtaatacaa accgaaaatg ttgaaagtat tagttaaagt
     6121 ggttatgcag tttttgcatt tatatatctg ttaatagatc aaaaatcatc gcttcgctga
     6181 ttaattaccc cagaaataag gctaaaaaac taatcgcatt atcatcctat ggttgttaat
     6241 ttgattcgtt catttgaagg tttgtggggc caggttactg ccaatttttc ctcttcataa
     6301 ccataaaagc tagtattgta gaatctttat tgttcggagc agtgcggcgc gaggcacatc
     6361 tgcgtttcag gaacgcgacc ggtgaagacg aggacgcacg gaggagagtc ttccttcgga
     6421 gggctgtcac ccgctcggcg gcttctaatc cgtacttcaa tatagcaatg agcagttaag
     6481 cgtattactg aaagttccaa agagaaggtt tttttaggct aagataatgg ggctctttac
     6541 atttccacaa catataagta agattagata tggatatgta tatggatatg tatatggtgg
     6601 taatgccatg taatatgatt attaaacttc tttgcgtcca tccaaaaaaa aagtaagaat
     6661 ttttgaaaat tcaaggaatt cgatatcaag cttatcgata ccgtcgacct cgaggggggg
     6721 cccggtacca gcttttgttc cctttagtga gggttaattt cgagcttggc gtaatcatgg
     6781 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     6841 ggaagcataa a
//
