LOCUS       sequence-8709-ex        7765 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 7765)
  AUTHORS   Dong J, Bloom JD, Goncharov V, Chattopadhyay M, Millhauser GL, Lynn 
            DG, Scheibel T, Lindquist S
  TITLE     Probing the role of PrP repeats in conformational conversion and 
            amyloid assembly of chimeric yeast prions.
  JOURNAL   J Biol Chem. 2007 Nov 23. 282(47):34204-12.
  PUBMED    17893150
REFERENCE   2  (bases 1 to 7765)
  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: "J Biol 
            Chem. 2007 Nov 23. 282(47):34204-12."
COMMENT     Sequence Label: pRS306Sup35R1+5
FEATURES             Location/Qualifiers
     source          1..7765
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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     2187..2209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2206..2222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2243..2261
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2290..2306
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2291..2307
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5773..5792)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5774..5792)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5799..5816)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5799..5815)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5808..5830)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5960..6415
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6047..6066)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6257..6278
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6546..7349)
                     /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(7350..7570)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7596..7615)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7715..7737
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttac ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctc ggaattaacc ctcactaaag ggaacaaaag ctgggtaccg
     2281 ggccccccct cgaggtcgac ggtatcgata agcttgatat cgaattcctc gagaagatat
     2341 ccatcatatt accattgtaa tactgccagg aataatagga ccaaccaaca ggcaaacagt
     2401 ttaaacgact tattaaccgc gttaataggt gttccaccat atgaaggcac tgaaccagaa
     2461 attgaagcaa ataccgaaca ggagggcgaa aagggagaag aaaaggataa gaaggataag
     2521 tctgaagcac ccaaagagga agctggcgaa accaacaaag aaaaaccttt gaatcagctg
     2581 gaggaatcgt cgagaccacc attagccaaa aaatcttcat cgttcgctca cctacaagcg
     2641 ccttccccaa tacctgaccc gttacaagta tccaagcctg aaacgagaat ggacttacca
     2701 ttttcaccag aagtgaatgt ctatgatacc gaggacactt acgtagttgt tcttgcgtta
     2761 ccaggtgcta actctagggc tttccacatt gattaccatc catcttctca tgagatgctc
     2821 atcaagggta agatcgaaga cagagtgggc attgatgaaa aattcttgaa gatcacggaa
     2881 ctaaaatatg gtgcgtttga gagaaccgtt aaattccccg tgctgccacg cattaaggac
     2941 gaagaaatta aagctactta caacaacggt ctactacaaa ttaaggtgcc taaaattgtc
     3001 aatgacactg aaaagccgaa gccaaaaaag aggatcgcca ttgaggaaat acccgacgaa
     3061 gaattggagt ttgaagaaaa tcccaaccct acggtagaaa attgaatatc gtatctgttt
     3121 atacacacat acatacattt atatttataa taagcgttaa aatttcggca gaatatctgt
     3181 caaccacaca aaaatcatac aacgaatggt atatgcttca tttctttgtt tcgcattagc
     3241 tgcgctattt gactcaaatt attatttttt actaagacga cgcgtcacag tgttcgagtc
     3301 tgtgtcattt cttttgtaat tctcttaaac cacttcataa agttgtgaag ttcatagcaa
     3361 aattcttccg caaaaagatg aatcttagtt ctcagcccac caaaagaggt acatgctaag
     3421 atcatacaga agttattgtc acttcttacc ttgctcttaa atgtacatta caaccgggta
     3481 ttatatctta catcatcgta taatatgatc tttctttatg gagaaaattt ttttttcact
     3541 cgaccaaagc tcccattgct tctgaagagt gtagtgtata ttggtacatc ttctcttgaa
     3601 agactccatt gtactgtaac aaaaagcggt ttcttcatcg acttgctcgg aataacatct
     3661 atatctgccc actagcaaca ggatccatgt cggattcaaa ccaaggcaac aatcagcaaa
     3721 actaccagca atacagccag aacggtaacc aacaacaagg taacaacaga taccaaggtt
     3781 atcaagctta caatgctcaa gcccaacctg caggtgggta ctaccaaaat taccaaggtt
     3841 attccggata tccacaaggt ggaggtactt ggggtcaacc ccatggagga ggttggggtc
     3901 aaccccatgg aggaggttgg ggtcaacccc atggaggtgg ttggggtcaa ccccatggag
     3961 gtggttgggg tcaaccacaa ggcggccgtg gaaattacaa aaacttcaac tacaataaca
     4021 atttgcaagg atatcaagct ggtttccaac cacagtctca aggtatgtct ttgaacgact
     4081 ttcaaaagca acaaaagcag gccgctccca aaccaaagaa gactttgaag cttgtctcca
     4141 gttccggtat caagttggcc aatgctacca agaaggttgg cacaaaacct gccgaatctg
     4201 ataagaaaga ggaagagaag tctgctgaaa ccaaagaacc aactaaagag ccaacaaagg
     4261 tcgaagaacc agttaaaaag gaggagaaac cagtccagac tgaagaaaag acggaggaaa
     4321 aatcggaact tccaaaggta gaagacctta aaatctctga atcaacacat aataccaaca
     4381 atgccaatgt taccagtgct gatgccttga tcaaggaaca ggaagaagaa gtggatgacg
     4441 aagttgttaa cgatccgcgg atgtttggtg gtaaagatca cgtttcttta attttcatgg
     4501 gtcatgttga tgccggtaaa tctactatgg gtggtaatct actatacttg actggctctg
     4561 tggataagag aactattgag aaatatgaaa gagaagccaa ggatgcaggc agacaaggtt
     4621 ggtacttgtc atgggtcatg gataccaaca aagaagaaag aaatgatggt aagactatcg
     4681 aagttggtaa ggcctacttt gaaactgaaa aaaggcgtta taccatattg gatgctcctg
     4741 gtcataaaat gtacgtttcc gagatgatcg gtggtgcttc tcaagctgat gttggtgttt
     4801 tggtcatttc cgccagaaag ggtgagtacg aaaccggttt tgagagaggt ggtcaaactc
     4861 gtgaacacgc cctattggcc aagacccaag gtgttaataa gatggttgtc gtcgtaaata
     4921 agatggatga cccaaccgtt aactggtcta aggaacgtta cgaccaatgt gtgagtaatg
     4981 tcagcaattt cttgagagca attggttaca acattaagac agacgttgta tttatgccag
     5041 tatccggcta cagtggtgca aatttgaaag atcacgtaga tccaaaagaa tgcccatggt
     5101 acaccggccc aactctgtta gaatatctgg atacaatgaa ccacgtcgac cgtcacatca
     5161 atgctccatt catgttgcct attgccgcta agatgaagga tctaggtacc atcgttgaag
     5221 gtaaaattga atccggtcat atcaaaaagg gtcaatccac cctactgatg cctaacaaaa
     5281 ccgctgtgga aattcaaaat atttacaacg aaactgaaaa tgaagttgat atggctatgt
     5341 gtggtgagca agttaaacta agaatcaaag gtgttgaaga agaagacatt tcaccaggtt
     5401 ttgtactaac atcgccaaag aaccctatca agagtgttac caagtttgta gctcaaattg
     5461 ctattgtaga attaaaatct atcatagcag ccggtttttc atgtgttatg catgttcata
     5521 cagcaattga agaggtacat attgttaagt tattgcacaa attagaaaag ggtaccaacc
     5581 gtaagtcaaa gaaaccacct gcttttgcta agaagggtat gaaggtcatc gctgttttag
     5641 aaactgaagc tccagtttgt gtggaaactt accaagatta ccctcaatta ggtagattca
     5701 ctttgagaga tcaaggtacc acaatagcaa ttggtaaaat tgttaaaatt gccgagtaag
     5761 agctccaatt cgccctatag tgagtcgtat tacaattcac tggccgtcgt tttacaacgt
     5821 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     5881 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     5941 ctgaatggcg aatggcgcga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     6001 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     6061 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     6121 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     6181 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     6241 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     6301 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     6361 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aatttcctga
     6421 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcataggg taataactga
     6481 tataattaaa ttgaagctct aatttgtgag tttagtatac atgcatttac ttataataca
     6541 gttttttagt tttgctggcc gcatcttctc aaatatgctt cccagcctgc ttttctgtaa
     6601 cgttcaccct ctaccttagc atcccttccc tttgcaaata gtcctcttcc aacaataata
     6661 atgtcagatc ctgtagagac cacatcatcc acggttctat actgttgacc caatgcgtct
     6721 cccttgtcat ctaaacccac accgggtgtc ataatcaacc aatcgtaacc ttcatctctt
     6781 ccacccatgt ctctttgagc aataaagccg ataacaaaat ctttgtcgct cttcgcaatg
     6841 tcaacagtac ccttagtata ttctccagta gatagggagc ccttgcatga caattctgct
     6901 aacatcaaaa ggcctctagg ttcctttgtt acttcttctg ccgcctgctt caaaccgcta
     6961 acaatacctg ggcccaccac accgtgtgca ttcgtaatgt ctgcccattc tgctattctg
     7021 tatacacccg cagagtactg caatttgact gtattaccaa tgtcagcaaa ttttctgtct
     7081 tcgaagagta aaaaattgta cttggcggat aatgccttta gcggcttaac tgtgccctcc
     7141 atggaaaaat cagtcaagat atccacatgt gtttttagta aacaaatttt gggacctaat
     7201 gcttcaacta actccagtaa ttccttggtg gtacgaacat ccaatgaagc acacaagttt
     7261 gtttgctttt cgtgcatgat attaaatagc ttggcagcaa caggactagg atgagtagca
     7321 gcacgttcct tatatgtagc tttcgacatg atttatcttc gtttcctgca ggtttttgtt
     7381 ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt cttcaacact acatatgcgt
     7441 atatatacca atctaagtct gtgctccttc cttcgttctt ccttctgttc ggagattacc
     7501 gaatcaaaaa aatttcaaag aaaccgaaat caaaaaaaag aataaaaaaa aaatgatgaa
     7561 ttgaattgaa aagctgtggt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     7621 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     7681 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     7741 tcaccgtcat caccgaaacg cgcga
//
