LOCUS       sequence-214327-        8945 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 8945)
  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 8945)
  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: 5538 pUG2-SSA1
FEATURES             Location/Qualifiers
     source          1..8945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2958..2974)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2958..2974)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2971..2993)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2982..2998
                     /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(3006..3012,3013..3030,3031..3036))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3051..3072
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3189..3206)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3360..3948)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3440..3459)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4119..4910,4911..4979))
                     /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     4742..4761
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4980..5084)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5152..5170
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      5528..6870
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    5860..5907
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        7462..7677
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             7678..8481
                     /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"
ORIGIN
        1 atgtcaaaag ctgtcggtat tgatttaggt acaacatact cgtgtgttgc tcactttgct
       61 aatgatcgtg tggacattat tgccaacgat caaggtaaca gaaccactcc atcttttgtc
      121 gctttcactg acactgaaag attgattggt gatgctgcta agaatcaagc tgctatgaat
      181 ccttcgaata ccgttttcga cgctaagcgt ttgatcggta gaaacttcaa cgacccagaa
      241 gtgcaggctg acatgaagca cttcccattc aagttgatcg atgttgacgg taagccacaa
      301 attcaagttg aatttaaggg tgaaaccaag aactttaccc cagaacaaat ctcctccatg
      361 gtcttgggta agatgaagga aactgccgaa tcttacttgg gtgccaaggt caatgacgct
      421 gtcgtcactg tcccagctta cttcaacgat tctcaaagac aagctaccaa ggatgctggt
      481 accattgctg gtttgaatgt cttgcgtatt attaacgaac ctaccgccgc tgccattgct
      541 tacggtttgg acaagaaggg taaggaagaa cacgtcttga ttttcgactt gggtggtggt
      601 actttcgatg tctctttgtt gtccattgaa gacggtatct ttgaagttaa ggccaccgct
      661 ggtgacaccc atttgggtgg tgaagatttt gacaacagat tggtcaacca cttcatccaa
      721 gaattcaaga gaaagaacaa gaaggacttg tctaccaacc aaagagcttt gagaagatta
      781 agaaccgctt gtgaaagagc caagagaact ttgtcttcct ccgctcaaac ttccgttgaa
      841 attgactctt tgttcgaagg tatcgatttc tacacttcca tcaccagagc cagattcgaa
      901 gaattgtgtg ctgacttgtt cagatctact ttggacccag ttgaaaaggt cttgagagat
      961 gctaaattgg acaaatctca agtcgatgaa attgtcttgg tcggtggttc taccagaatt
     1021 ccaaaggtcc aaaaattggt cactgactac ttcaacggta aggaaccaaa cagatctatc
     1081 aacccagatg aagctgttgc ttacggtgct gctgttcaag ctgctatttt gactggtgac
     1141 gaatcttcca agactcaaga tctattgttg ttggatgtcg ctccattatc cttgggtatt
     1201 gaaactgctg gtggtgtcat gaccaagttg attccaagaa actctaccat tccaacaaag
     1261 aagtccgaga tcttttccac ttatgctgat aaccaaccag gtgtcttgat tcaagtcttt
     1321 gaaggtgaaa gagccaagac taaggacaac aacttgttgg gtaagttcga attgagtggt
     1381 attccaccag ctccaagagg tgtcccacaa attgaagtca ctttcgatgt cgactctaac
     1441 ggtattttga atgtttccgc cgtcgaaaag ggtactggta agtctaacaa gatcactatt
     1501 accaacgaca agggtagatt gtccaaggaa gatatcgaaa agatggttgc tgaagccgaa
     1561 aaattcaagg aagaagatga aaaggaatct caaagaattg cttccaagaa ccaattggaa
     1621 tccattgctt actctttgaa gaacaccatt tctgaagctg gtgacaaatt ggaacaagct
     1681 gacaaggaca ccgtcaccaa gaaggctgaa gagactattt cttggttaga cagcaacacc
     1741 actgccagca aggaagaatt cgatgacaag ttgaaggagt tgcaagacat tgccaaccca
     1801 atcatgtcta agttgtacca agctggtggt gctccaggtg gcgctgcagg tggtgctcca
     1861 ggcggtttcc caggtggtgc tcctccagct ccagaggctg aaggtccaac cgttgaagaa
     1921 gttgattaag ccaattggtg cggcaattga taataacgaa aatgtctttt aatgatctgg
     1981 gtataatgag gaattttccg aacgttttta ctttatatat atatatacat gtaacatata
     2041 ttctatacgc tatagagaaa ggaaattttt caattaaaaa aaaaatagag aaagagtttc
     2101 acttcttgat tatcgctaac actaatggtt gaagtactgc tactttaatt ttatagatag
     2161 gcaaaaaaaa attattcggg gcgagctggg aattgaaccc agggcctctc gcatgcacca
     2221 ttccttgcgg cggcggtgct caacggcctc aacctactac tgggctgctt cctaatgcag
     2281 gagtcgcata agggagagcg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc
     2341 ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg
     2401 caactcgtag gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc
     2461 tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca cgccctcgct
     2521 caagccttcg tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc
     2581 ggcatggcgg ccgacgcgct gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg
     2641 gccttcccca ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc
     2701 atgctgtcca ggcaggtaga tgacgaccat cagggacagc ttcaaggatc gctcgcggct
     2761 cttaccagcc taacttcgat cattggaccg ctgatcgtca cggcgattta tgccgcctcg
     2821 gcgagcacat ggaacgggtt ggcatggatt gtaggcgccg ccctatacct tgtctgcctc
     2881 cccgcgttgc gtcgcggtgc atggagccgg gccacctact gaagtggatt tctttaagag
     2941 ctcgaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     3001 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     3061 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     3121 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     3181 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     3241 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     3301 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3361 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3421 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3481 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3541 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3601 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3661 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3721 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3781 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3841 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3901 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3961 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4021 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     4081 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     4141 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     4201 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     4261 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     4321 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4381 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     4441 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     4501 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     4561 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     4621 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     4681 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     4741 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4801 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4861 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4921 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4981 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     5041 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     5101 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     5161 atcacgaggc cctttcgtct tcaagaattc tgaaccagtc ctaaaacgag taaataggac
     5221 cggcaattct tcaagcaata aacaggaata ccaattatta aaagataact tagtcagatc
     5281 gtacaataaa gctttgaaga aaaatgcgcc ttattcaatc tttgctataa aaaatggccc
     5341 aaaatctcac attggaagac atttgatgac ctcatttctt tcaatgaagg gcctaacgga
     5401 gttgactaat gttgtgggaa attggagcga taagcgtgct tctgccgtgg ccaggacaac
     5461 gtatactcat cagataacag caatacctga tcactacttc gcactagttt ctcggtacta
     5521 tgcatatgat ccaatatcaa aggaaatgat agcattgaag gatgagacta atccaattga
     5581 ggagtggcag catatagaac agctaaaggg tagtgctgaa ggaagcatac gataccccgc
     5641 atggaatggg ataatatcac aggaggtact agactacctt tcatcctaca taaatagacg
     5701 catataagta cgcatttaag cataaacacg cactatgccg ttcttctcat gtatatatat
     5761 atacaggcaa cacgcagata taggtgcgac gtgaacagtg agctgtatgt gcgcagctcg
     5821 cgttgcattt tcggaagcgc tcgttttcgg aaacgctttg aagttcctat tccgaagttc
     5881 ctattctcta gaaagtatag gaacttcaga gcgcttttga aaaccaaaag cgctctgaag
     5941 acgcactttc aaaaaaccaa aaacgcaccg gactgtaacg agctactaaa atattgcgaa
     6001 taccgcttcc acaaacattg ctcaaaagta tctctttgct atatatctct gtgctatatc
     6061 cctatataac ctacccatcc acctttcgct ccttgaactt gcatctaaac tcgacctcta
     6121 cattttttat gtttatctct agtattactc tttagacaaa aaaattgtag taagaactat
     6181 tcatagagtg aatcgaaaac aatacgaaaa tgtaaacatt tcctatacgt agtatataga
     6241 gacaaaatag aagaaaccgt tcataatttt ctgaccaatg aagaatcatc aacgctatca
     6301 ctttctgttc acaaagtatg cgcaatccac atcggtatag aatataatcg gggatgcctt
     6361 tatcttgaaa aaatgcaccc gcagcttcgc tagtaatcag taaacgcggg aagtggagtc
     6421 aggctttttt tatggaagag aaaatagaca ccaaagtagc cttcttctaa ccttaacgga
     6481 cctacagtgc aaaaagttat caagagactg cattatagag cgcacaaagg agaaaaaaag
     6541 taatctaaga tgctttgtta gaaaaatagc gctctcggga tgcatttttg tagaacaaaa
     6601 aagaagtata gattctttgt tggtaaaata gcgctctcgc gttgcatttc tgttctgtaa
     6661 aaatgcagct cagattcttt gtttgaaaaa ttagcgctct cgcgttgcat ttttgtttta
     6721 caaaaatgaa gcacagattc ttcgttggta aaatagcgct ttcgcgttgc atttctgttc
     6781 tgtaaaaatg cagctcagat tctttgtttg aaaaattagc gctctcgcgt tgcatttttg
     6841 ttctacaaaa tgaagcacag atgcttcgtt aacaaagata tgctattgaa gtgcaagatg
     6901 gaaacgcaga aaatgaaccg gggatgcgac gtgcaagatt acctatgcaa tagatgcaat
     6961 agtttctcca ggaaccgaaa tacatacatt gtcttccgta aagcgctaga ctatatatta
     7021 ttatacaggt tcaaatatac tatctgtttc agggaaaact cccaggttcg gatgttcaaa
     7081 attcaatgat gggtaacaag tacgatcgta aatctgtaaa acagtttgtc ggatattagg
     7141 ctgtatctcc tcaaagcgta ttcgaatatc attgagaagc tgcagcgtca catcggataa
     7201 taatgatggc agccattgta gaagtgcctt ttgcatttct agtctctttc tcggtctagc
     7261 tagttttact acatcgcgaa gatagaatct tagatcacac tgcctttgct gagctggatc
     7321 aatagagtaa caaaagagtg gtaaggcctc gttaaaggac aaggacctga gcggaagtgt
     7381 atcgtacagt agacggagta tactagtata gtctatagtc cgtggaattc tcatgtttga
     7441 cagcttatca tcgataagct tttcaattca tcattttttt tttattcttt tttttgattt
     7501 cggtttcctt gaaatttttt tgattcggta atctccgaac agaaggaaga acgaaggaag
     7561 gagcacagac ttagattggt atatatacgc atatgtagtg ttgaagaaac atgaaattgc
     7621 ccagtattct taacccaact gcacagaaca aaaacctgca ggaaacgaag ataaatcatg
     7681 tcgaaagcta catataagga acgtgctgct actcatccta gtcctgttgc tgccaagcta
     7741 tttaatatca tgcacgaaaa gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc
     7801 aaggaattac tggagttagt tgaagcatta ggtcccaaaa tttgtttact aaaaacacat
     7861 gtggatatct tgactgattt ttccatggag ggcacagtta agccgctaaa ggcattatcc
     7921 gccaagtaca attttttact cttcgaagac agaaaatttg ctgacattgg taatacagtc
     7981 aaattgcagt actctgcggg tgtatacaga atagcagaat gggcagacat tacgaatgca
     8041 cacggtgtgg tgggcccagg tattgttagc ggtttgaagc aggcggcaga agaagtaaca
     8101 aaggaaccta gaggcctttt gatgttagca gaattgtcat gcaagggctc cctatctact
     8161 ggagaatata ctaagggtac tgttgacatt gcgaagagcg acaaagattt tgttatcggc
     8221 tttattgctc aaagagacat gggtggaaga gatgaaggtt acgattggtt gattatgaca
     8281 cccggtgtgg gtttagatga caagggagac gcattgggtc aacagtatag aaccgtggat
     8341 gatgtggtct ctacaggatc tgacattatt attgttggaa gaggactatt tgcaaaggga
     8401 agggatgcta aggtagaggg tgaacgttac agaaaagcag gctgggaagc atatttgaga
     8461 agatgcggcc agcaaaacta aaaaactgta ttataagtaa atgcatgtat actaaactca
     8521 caaattagag cttcaattta attatatcag ttattaccct cgacctcgac tgtacaggat
     8581 gttctagcta ctcgagtagc tagaacatcc tgtacagtcg agtagctaga acatcctgta
     8641 cagtcgagca gatccgccag gcgtgtatat agcgtggatg gccaggcaac tttagtgctg
     8701 acacatacag gcatatatat atgtgtgcga cgacacatga tcatatggca tgcatgtgct
     8761 ctgtatgtat ataaaactct tgttttcttc ttttctctaa atattctttc cttatacatt
     8821 aggtcctttg tagcataaat tactatactt ctatagacac gcaaacacgg atccaagctt
     8881 ggcacatcaa aagaaaagta atcaagtatt acaagaaaca aaaattcaag taaataacag
     8941 ataat
//
