LOCUS       sequence-494941-        9437 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 9437)
  AUTHORS   Glover JR, Lindquist S
  TITLE     Hsp104, Hsp70, and Hsp40: a novel chaperone system that rescues 
            previously aggregated proteins.
  JOURNAL   Cell 1998 Jul 10;94(1):73-82.
  PUBMED    9674429
REFERENCE   2  (bases 1 to 9437)
  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: "Cell"; 
            date: "1998-07-10"; volume: "94(1)"; pages: "73-82"
COMMENT     Sequence Label: 6379 H10-K218T
FEATURES             Location/Qualifiers
     source          1..9437
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     154..171
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    288..309
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(324..329,330..347,348..354)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    362..378
                     /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     367..389
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     386..402
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     386..402
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        423..441
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     469..485
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     470..486
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5075..5091)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5075..5091)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5123..5142)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5124..5142)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5149..5166)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5149..5165)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5158..5180)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5306..5761
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5393..5412)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5603..5624
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5892..6695)
                     /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(6696..6911)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6935..6954)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7054..7076
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7114..7132)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    7173..7676
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        7713..7817
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7818..7886,7887..8678)
                     /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(8036..8055)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8849..9437
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9338..9357
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      121 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      181 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      241 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      301 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      361 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc
      421 gaaattaacc ctcactaaag ggaacaaaag ctggtaccgg gccccccctc gaggtcgacg
      481 gtatcgattc aaaggcgtta ttcagcatca ttggaagtga aatggactgg gtcgacgaca
      541 agttggcctc taagtttgtc ttcaagaatc caaactccaa gggcacatgc ggttgtggcg
      601 agagtttcat ggtttaaaaa ccttctgcac catttttaga aaaaaagaat ctacctattc
      661 acttatttat tcatttactt atttatttac atatttatca tacatattaa cattgaaccc
      721 tccatcgtgg tagtgtttgc tgttcctaac ttttctttcg ttgttcttgt agatatatat
      781 ttttccagaa ttttctagaa gggttattaa ttacaatctt aaacgttcca taaggggccg
      841 cgattttttt gttcaatttt caacaggggg cccatctcaa agaactgcaa attatatcac
      901 agtaaaaggc aaaggggcgc aaacttatgc aacctgccag attattatat aaggcattgt
      961 aatcttgcct caattccttc ataattcgtt cctttgtcac ttgttccttt ttacccttga
     1021 atcgaatcag caataacaaa gaaaaaagaa atcaactaca cgtaccataa aatatacaga
     1081 atatatgaac gaccaaacgc aatttacaga aagggctcta acgattttga cgttggctca
     1141 aaaattggct tcggatcatc aacatccaca attacaacct atacatattc tagctgcctt
     1201 cattgaaacg ccagaagatg gatcagtccc ttacctacag aatctaattg agaagggccg
     1261 ttacgactat gatcttttca agaaagtggt taatagaaat ctagtaagaa ttcctcaaca
     1321 gcaacctgca cctgcggaga taactccaag ttatgctttg gggaaagtcc ttcaagacgc
     1381 tgctaagatt caaaaacaac agaaggactc atttatagcg caagaccata tattgtttgc
     1441 tctattcaat gattcgtcta ttcagcaaat atttaaggaa gctcaagtag atattgaggc
     1501 catcaagcaa caagctcttg aacttcgtgg taacactaga attgactctc gtggcgctga
     1561 tacgaacaca cctttggaat atttatcaaa gtacgccatt gatatgactg agcaggctcg
     1621 tcaaggtaaa cttgaccctg tcatcggccg tgaagaagaa ataagaagca ctattagagt
     1681 tttagcaaga agaattaagt ccaacccatg tttaattggt gagccaggta tcggtaccac
     1741 cgctattatt gaaggtgttg ctcaaagaat cattgacgat gacgttccca ctatcttaca
     1801 aggcgctaaa ttgttcagtc tagatttggc cgcattaacc gcaggtgcta aatacaaagg
     1861 tgatttcgaa gaaagattca aaggtgtttt gaaggaaatc gaagaatcaa agactctaat
     1921 tgtgttattc attgatgaaa ttcacatgtt aatgggtaat ggtaaggacg acgctgctaa
     1981 catcttgaag ccagctttgt ccagaggcca attgaaggtc atcggtgcca ccaccaataa
     2041 cgaatataga tctattgtgg aaaaggatgg tgcctttgaa agaagattcc agaaaattga
     2101 agtcgctgaa ccaagtgtga gacaaacagt ggccatattg agaggtctgc aaccaaagta
     2161 tgaaatacat catggtgtaa ggattctgga tagcgcctta gtcactgctg ctcaattagc
     2221 caagcgttac ttgccatata gaagattgcc agattctgct ttggatttag ttgatatttc
     2281 ttgtgctggt gtcgccgtcg caagagattc taagccagaa gaattggatt ccaaggaacg
     2341 tcaattgcaa ttgattcaag tagagataaa agctctagag agagatgaag atgccgactc
     2401 caccactaaa gatagattaa agttagctag gcagaaggaa gcttcattgc aagaagaatt
     2461 ggaacctcta agacaacgtt acaatgaaga aaagcatggc catgaagaat tgacacaagc
     2521 taaaaagaaa ttggatgaac tggaaaacaa ggcccttgat gctgaacgta gatatgatac
     2581 tgctaccgcc gctgatttaa ggtacttcgc catcccagat atcaaaaagc aaatcgaaaa
     2641 gcttgaagat caggttgctg aggaagagag acgtgctggt gccaactcca tgatccaaaa
     2701 tgtggtcgat tcagacacca tttctgaaac agctgcaaga ttgactggta tccctgttaa
     2761 gaagttgtca gaatctgaaa atgaaaaatt gattcatatg gaacgtgact tatcatctga
     2821 agtcgtgggc caaatggatg ccattaaagc tgtttccaat gccgttagat tgtctagatc
     2881 aggtttagct aatccaaggc aaccagcatc cttcttattt ttaggtttgt ccggttccgg
     2941 taaaactgaa ttggctaaaa aagttgctgg atttttgttt aatgatgagg acatgatgat
     3001 cagggtcgat tgttctgaat taagcgagaa gtatgcggtc tctaagttgt tgggtaccac
     3061 ggcaggttat gtcgggtacg atgaaggtgg ctttttaact aaccaactgc aatacaaacc
     3121 atactccgtt ttgttattcg atgaagtaga aaaggcacat cctgatgttt tgactgtcat
     3181 gctacaaatg ttggatgacg gtagaattac ttctggtcaa ggtaagacga tcgactgttc
     3241 caattgtatt gtcatcatga cttccaatct aggtgctgaa tttatcaatt ctcaacaagg
     3301 atcaaagatc caagaatcta ccaagaattt ggtcatgggt gctgttaggc aacatttcag
     3361 accagaattt ttgaacagaa tttctagtat agtcattttc aacaagctat ctagaaaagc
     3421 tattcataag atcgtggata ttcgtttgaa ggaaattgaa gagagattcg agcaaaatga
     3481 taaacattac aagttgaatt taactcaaga ggccaaggac ttcttggcca aatatggtta
     3541 ttccgatgat atgggtgcac gtccactgaa caggttaatt caaaacgaaa ttttgaacaa
     3601 actggcacta aggatcttaa agaatgaaat caaggataag gaaactgtca atgtcgtctt
     3661 gaagaagggt aaatctcgtg atgaaaatgt tcctgaggaa gctgaagaat gtctggaagt
     3721 tctaccaaat cacgaagcta ctataggggc tgacacgtta ggtgatgacg ataatgagga
     3781 cagtatggaa attgatgatg acctagatta atttaatata gtgtgatttt taaaaacttt
     3841 cgaacaagaa tcagtaatat aatatatata attaataaaa ctaatggaat ttgtttaatt
     3901 gaacttgaca cccgagcaaa agagctcaca ttttaacgtt tagttcatct tgctgtgaat
     3961 aatctaaaag taaacaatgg acaaaaaagt ccactggacg tcaaatcttt ttttattgta
     4021 tttgggacat atggcgttga taactaaagt ttaaaggaag taataagtga aggttaaatc
     4081 tggacctctt tactttcata ttagaagtat aaaacacacg cggattggtc cattattaga
     4141 atacgggtac tacaataatg gttttgatag ttactgtaca tttgaatgta gcgactcata
     4201 tgactcatat atatgactac caaatcatgt taaaattagt atatctttaa cttagacttc
     4261 agtactatct gaactgcacg tttaaatttt taccatagca gttcatgtaa tattaataat
     4321 cacttgtgcc aatataatgt tacagcgaac atttttccat gcgcttggaa aattgaatga
     4381 tagaaaattc gaacagcgat gatttccaac gatgtaatcc attaagatga ttgttagaat
     4441 ttctattgtt gcagaggtca tatgatatta agaacacagc acaatatatt aaaatttctt
     4501 ctcgaggata ttgagaaatg ggtgaatttt gaggtaattg ttgggattcc attgttaata
     4561 agggctataa tattaggtat acagaatata ctagatgttc tcctcgagga tctacgaatc
     4621 ctcataatgg aatcaatatt tctatatact aatattacga ttatccctca ttccgtttta
     4681 tatgtttcat tatcttatta cattatcaat cctttcacta cagcttcctc taacctcgat
     4741 gacagcttct cataacctat gtcatcatct aacaccgtat atgataatat agtgatagtt
     4801 ataactatta gttgatagac gatagtggat ttttattcca acagatataa gaattacaaa
     4861 aagagaactg ttatttctac ataataatat tacgattagt actcctttta tatgtttctt
     4921 tatccattta cattatcaat cctgattaaa attcccacga acccccagta ctagattgtt
     4981 cataatggat caagttcaca agttaataag ggcgtgtggt ctagtggtat gattctcgct
     5041 ttgggcgact tcctgactaa acaggaagac aaaggatcca ctagttctag agcggccgcc
     5101 accgcggtgg agctccaatt cgccctatag tgagtcgtat tacaattcac tggccgtcgt
     5161 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     5221 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     5281 gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     5341 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     5401 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     5461 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     5521 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     5581 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     5641 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     5701 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     5761 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagggtaat
     5821 aactgatata attaaattga agctctaatt tgtgagttta gtatacatgc atttacttat
     5881 aatacagttt tttagttttg ctggccgcat cttctcaaat atgcttccca gcctgctttt
     5941 ctgtaacgtt caccctctac cttagcatcc cttccctttg caaatagtcc tcttccaaca
     6001 ataataatgt cagatcctgt agagaccaca tcatccacgg ttctatactg ttgacccaat
     6061 gcgtctccct tgtcatctaa acccacaccg ggtgtcataa tcaaccaatc gtaaccttca
     6121 tctcttccac ccatgtctct ttgagcaata aagccgataa caaaatcttt gtcgctcttc
     6181 gcaatgtcaa cagtaccctt agtatattct ccagtagata gggagccctt gcatgacaat
     6241 tctgctaaca tcaaaaggcc tctaggttcc tttgttactt cttctgccgc ctgcttcaaa
     6301 ccgctaacaa tacctgggcc caccacaccg tgtgcattcg taatgtctgc ccattctgct
     6361 attctgtata cacccgcaga gtactgcaat ttgactgtat taccaatgtc agcaaatttt
     6421 ctgtcttcga agagtaaaaa attgtacttg gcggataatg cctttagcgg cttaactgtg
     6481 ccctccatgg aaaaatcagt caagatatcc acatgtgttt ttagtaaaca aattttggga
     6541 cctaatgctt caactaactc cagtaattcc ttggtggtac gaacatccaa tgaagcacac
     6601 aagtttgttt gcttttcgtg catgatatta aatagcttgg cagcaacagg actaggatga
     6661 gtagcagcac gttccttata tgtagctttc gacatgattt atcttcgttt cctgcaggtt
     6721 tttgttctgt gcagttgggt taagaatact gggcaatttc atgtttcttc aacactacat
     6781 atgcgtatat ataccaatct aagtctgtgc tccttccttc gttcttcctt ctgttcggag
     6841 attaccgaat caaaaaaatt tcaaggaaac cgaaatcaaa aaaaagaata aaaaaaaaat
     6901 gatgaattga aaaggtggta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     6961 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     7021 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     7081 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     7141 ttaatgtcat gataataatg gtttcttaga cggatcgctt gcctgtaact tacacgcgcc
     7201 tcgtatcttt taatgatgga ataatttggg aatttactct gtgtttattt atttttatgt
     7261 tttgtatttg gattttagaa agtaaataaa gaaggtagaa gagttacgga atgaagaaaa
     7321 aaaaataaac aaaggtttaa aaaatttcaa caaaaagcgt actttacata tatatttatt
     7381 agacaagaaa agcagattaa atagatatac attcgattaa cgataagtaa aatgtaaaat
     7441 cacaggattt tcgtgtgtgg tcttctacac agacaagatg aaacaattcg gcattaatac
     7501 ctgagagcag gaagagcaag ataaaaggta gtatttgttg gcgatccccc tagagtcttt
     7561 tacatcttcg gaaaacaaaa actatttttt ctttaatttc tttttttact ttctattttt
     7621 aatttatata tttatattaa aaaatttaaa ttataattat ttttatagca cgtgatgaaa
     7681 aggacccagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     7741 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     7801 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     7861 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     7921 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     7981 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     8041 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     8101 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     8161 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     8221 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     8281 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     8341 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     8401 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     8461 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     8521 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     8581 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     8641 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     8701 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     8761 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     8821 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     8881 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     8941 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     9001 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     9061 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     9121 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     9181 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     9241 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     9301 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     9361 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     9421 ggcggagcct atggaaa
//
