LOCUS       sequence-177701-        6451 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 6451)
  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 6451)
  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: CDC12(K351N)
FEATURES             Location/Qualifiers
     source          1..6451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(217..236)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     336..358
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(396..414)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    455..958
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        995..1099
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1100..1168,1169..1960)
                     /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(1318..1337)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2131..2719
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2620..2639
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2873..2890
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3007..3028
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3043..3048,3049..3066,3067..3073)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3081..3097
                     /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     3086..3108
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3105..3121
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3105..3121
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3142..3160
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     3188..3204
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3189..3205
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4856..4875)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4857..4875)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4882..4899)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4882..4898)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4891..4913)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5039..5494
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5126..5145)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5336..5357
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5625..6428)
                     /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"
     promoter        complement(join(6429..6451,1..193))
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
ORIGIN
        1 tttttgttct gtgcagttgg gttaagaata ctgggcaatt tcatgtttct tcaacactac
       61 atatgcgtat atataccaat ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg
      121 agattaccga atcaaaaaaa tttcaaggaa accgaaatca aaaaaaagaa taaaaaaaaa
      181 atgatgaatt gaaaaggtgg tatggtgcac tctcagtaca atctgctctg atgccgcata
      241 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
      301 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
      361 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
      421 ggttaatgtc atgataataa tggtttctta gacggatcgc ttgcctgtaa cttacacgcg
      481 cctcgtatct tttaatgatg gaataatttg ggaatttact ctgtgtttat ttatttttat
      541 gttttgtatt tggattttag aaagtaaata aagaaggtag aagagttacg gaatgaagaa
      601 aaaaaaataa acaaaggttt aaaaaatttc aacaaaaagc gtactttaca tatatattta
      661 ttagacaaga aaagcagatt aaatagatat acattcgatt aacgataagt aaaatgtaaa
      721 atcacaggat tttcgtgtgt ggtcttctac acagacaaga tgaaacaatt cggcattaat
      781 acctgagagc aggaagagca agataaaagg tagtatttgt tggcgatccc cctagagtct
      841 tttacatctt cggaaaacaa aaactatttt ttctttaatt tcttttttta ctttctattt
      901 ttaatttata tatttatatt aaaaaattta aattataatt atttttatag cacgtgatga
      961 aaaggaccca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     1021 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     1081 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     1141 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     1201 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     1261 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     1321 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     1381 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     1441 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     1501 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     1561 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     1621 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     1681 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     1741 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     1801 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     1861 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     1921 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     1981 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     2041 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     2101 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2161 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2221 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     2281 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2341 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     2401 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2461 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     2521 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     2581 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2641 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2701 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     2761 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     2821 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     2881 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     2941 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     3001 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     3061 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     3121 ccatgattac gccaagctcg aaattaaccc tcactaaagg gaacaaaagc tggtaccggg
     3181 ccccccctcg aggtcgacgg tatcgataag cttgatatcg aattcctgca gcccggggga
     3241 tccgatgcag tatcacgatt agcaattcag ctatgagtta tgttgctctt tgttttgttt
     3301 atggaaattg tcctatggta agtctctttt ttttgcaatc gtgattacag aaaaaaaaca
     3361 gggcgctgga aaagtgaaga atccgaaatt tttttcgaaa tcaccattgt ttgttttgag
     3421 tagatcaaag tcttgaaagg tgcagcaaga tataggatct tgacctgaag agtattgata
     3481 acgaactaca tcacatattg tatcaaataa tgagtgctgc cactgcgact gctgcccctg
     3541 ttcctcctcc tgtagggata tccaatcttc ccaaccagcg ttataaaatt gttaatgaag
     3601 aaggtggtac attcaccgtg atgttatgtg gtgagagcgg attgggtaaa actacattta
     3661 ttaatacctt atttcagact gtcttgaaaa gggcagatgg tcagcaacac cgccaagaac
     3721 ccatcagaaa gactgtagaa atcgacatca caagagcttt gctggaagag aagcatttcg
     3781 aattacgcgt taatgtcatc gacaccccag gttttggaga taacgtgaat aacaacaaag
     3841 cctggcaacc attggtggat tttattgatg atcaacacga ttcttatatg cgtcaagagc
     3901 agcaaccgta ccgtactaag aaatttgact tgagagttca tgccgttctg tacttcatca
     3961 ggcccactgg acatggttta aagccaattg acattgagac tatgaagaga ttgtcaacca
     4021 gagccaatct gattccagtt attgcgaaag ctgatacatt gactgcccaa gaattacaac
     4081 agttcaagtc cagaatcagg caagtaatag aagctcaaga aattcgtatt ttcacgcctc
     4141 cattagacgc tgattctaag gaagatgcca aaagtggcag taatccagat tctgccgctg
     4201 ttgaacatgc aagacagttg atagaagcaa tgccatttgc catagtaggg tctgaaaaaa
     4261 aatttgacaa tgggcaaggc actcaggtag ttgcaaggaa gtatccttgg ggacttgtgg
     4321 aaatcgaaaa cgattctcat tgtgatttcc gtaagctcag agccctgtta ctgaggactt
     4381 atctattaga tttgatttct accacccaag aaatgcatta cgaaacatac agaagattaa
     4441 gacttgaggg tcacgagaac accggggagg ggaatgaaga tttcactctt ccagcaattg
     4501 caccagcaag gaaattgtca cacaatccta gatacaaaga agaagaaaat gcattgaaga
     4561 actatttcac tgaccaagtc aaagctgagg agcaaagatt tagacaatgg gagcaaaaca
     4621 ttgttaacga aagaatcaga ctgaacggtg acttggaaga aattcaagga aaggtaaaaa
     4681 aactagaaga gcaggtcaaa agcttgcaag taaaaaaatc ccatttaaaa tgatgattaa
     4741 ttaatgtctt cctctttgtc tcgtcaattt caacgcctat ctctttgtct cttcttttgt
     4801 cgcagcaata cgcacatatc tagagcggcc gccaccgcgg tggagctcca attcgcccta
     4861 tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     4921 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     4981 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggac
     5041 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     5101 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     5161 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     5221 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     5281 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5341 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     5401 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5461 gcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc
     5521 atctgtgcgg tatttcacac cgcatagggt aataactgat ataattaaat tgaagctcta
     5581 atttgtgagt ttagtataca tgcatttact tataatacag ttttttagtt ttgctggccg
     5641 catcttctca aatatgcttc ccagcctgct tttctgtaac gttcaccctc taccttagca
     5701 tcccttccct ttgcaaatag tcctcttcca acaataataa tgtcagatcc tgtagagacc
     5761 acatcatcca cggttctata ctgttgaccc aatgcgtctc ccttgtcatc taaacccaca
     5821 ccgggtgtca taatcaacca atcgtaacct tcatctcttc cacccatgtc tctttgagca
     5881 ataaagccga taacaaaatc tttgtcgctc ttcgcaatgt caacagtacc cttagtatat
     5941 tctccagtag atagggagcc cttgcatgac aattctgcta acatcaaaag gcctctaggt
     6001 tcctttgtta cttcttctgc cgcctgcttc aaaccgctaa caatacctgg gcccaccaca
     6061 ccgtgtgcat tcgtaatgtc tgcccattct gctattctgt atacacccgc agagtactgc
     6121 aatttgactg tattaccaat gtcagcaaat tttctgtctt cgaagagtaa aaaattgtac
     6181 ttggcggata atgcctttag cggcttaact gtgccctcca tggaaaaatc agtcaagata
     6241 tccacatgtg tttttagtaa acaaattttg ggacctaatg cttcaactaa ctccagtaat
     6301 tccttggtgg tacgaacatc caatgaagca cacaagtttg tttgcttttc gtgcatgata
     6361 ttaaatagct tggcagcaac aggactagga tgagtagcag cacgttcctt atatgtagct
     6421 ttcgacatga tttatcttcg tttcctgcag g
//
