LOCUS       sequence-8708-ex        7741 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 7741)
  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 7741)
  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+4
FEATURES             Location/Qualifiers
     source          1..7741
                     /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(5749..5768)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5750..5768)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5775..5792)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5775..5791)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5784..5806)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5936..6391
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6023..6042)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6233..6254
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6522..7325)
                     /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(7326..7546)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7572..7591)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7691..7713
                     /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 ccacaaggcg
     3961 gccgtggaaa ttacaaaaac ttcaactaca ataacaattt gcaaggatat caagctggtt
     4021 tccaaccaca gtctcaaggt atgtctttga acgactttca aaagcaacaa aagcaggccg
     4081 ctcccaaacc aaagaagact ttgaagcttg tctccagttc cggtatcaag ttggccaatg
     4141 ctaccaagaa ggttggcaca aaacctgccg aatctgataa gaaagaggaa gagaagtctg
     4201 ctgaaaccaa agaaccaact aaagagccaa caaaggtcga agaaccagtt aaaaaggagg
     4261 agaaaccagt ccagactgaa gaaaagacgg aggaaaaatc ggaacttcca aaggtagaag
     4321 accttaaaat ctctgaatca acacataata ccaacaatgc caatgttacc agtgctgatg
     4381 ccttgatcaa ggaacaggaa gaagaagtgg atgacgaagt tgttaacgat ccgcggatgt
     4441 ttggtggtaa agatcacgtt tctttaattt tcatgggtca tgttgatgcc ggtaaatcta
     4501 ctatgggtgg taatctacta tacttgactg gctctgtgga taagagaact attgagaaat
     4561 atgaaagaga agccaaggat gcaggcagac aaggttggta cttgtcatgg gtcatggata
     4621 ccaacaaaga agaaagaaat gatggtaaga ctatcgaagt tggtaaggcc tactttgaaa
     4681 ctgaaaaaag gcgttatacc atattggatg ctcctggtca taaaatgtac gtttccgaga
     4741 tgatcggtgg tgcttctcaa gctgatgttg gtgttttggt catttccgcc agaaagggtg
     4801 agtacgaaac cggttttgag agaggtggtc aaactcgtga acacgcccta ttggccaaga
     4861 cccaaggtgt taataagatg gttgtcgtcg taaataagat ggatgaccca accgttaact
     4921 ggtctaagga acgttacgac caatgtgtga gtaatgtcag caatttcttg agagcaattg
     4981 gttacaacat taagacagac gttgtattta tgccagtatc cggctacagt ggtgcaaatt
     5041 tgaaagatca cgtagatcca aaagaatgcc catggtacac cggcccaact ctgttagaat
     5101 atctggatac aatgaaccac gtcgaccgtc acatcaatgc tccattcatg ttgcctattg
     5161 ccgctaagat gaaggatcta ggtaccatcg ttgaaggtaa aattgaatcc ggtcatatca
     5221 aaaagggtca atccacccta ctgatgccta acaaaaccgc tgtggaaatt caaaatattt
     5281 acaacgaaac tgaaaatgaa gttgatatgg ctatgtgtgg tgagcaagtt aaactaagaa
     5341 tcaaaggtgt tgaagaagaa gacatttcac caggttttgt actaacatcg ccaaagaacc
     5401 ctatcaagag tgttaccaag tttgtagctc aaattgctat tgtagaatta aaatctatca
     5461 tagcagccgg tttttcatgt gttatgcatg ttcatacagc aattgaagag gtacatattg
     5521 ttaagttatt gcacaaatta gaaaagggta ccaaccgtaa gtcaaagaaa ccacctgctt
     5581 ttgctaagaa gggtatgaag gtcatcgctg ttttagaaac tgaagctcca gtttgtgtgg
     5641 aaacttacca agattaccct caattaggta gattcacttt gagagatcaa ggtaccacaa
     5701 tagcaattgg taaaattgtt aaaattgccg agtaagagct ccaattcgcc ctatagtgag
     5761 tcgtattaca attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     5821 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     5881 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcgacgcg
     5941 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     6001 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     6061 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     6121 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     6181 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     6241 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     6301 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     6361 aattttaaca aaatattaac gtttacaatt tcctgatgcg gtattttctc cttacgcatc
     6421 tgtgcggtat ttcacaccgc atagggtaat aactgatata attaaattga agctctaatt
     6481 tgtgagttta gtatacatgc atttacttat aatacagttt tttagttttg ctggccgcat
     6541 cttctcaaat atgcttccca gcctgctttt ctgtaacgtt caccctctac cttagcatcc
     6601 cttccctttg caaatagtcc tcttccaaca ataataatgt cagatcctgt agagaccaca
     6661 tcatccacgg ttctatactg ttgacccaat gcgtctccct tgtcatctaa acccacaccg
     6721 ggtgtcataa tcaaccaatc gtaaccttca tctcttccac ccatgtctct ttgagcaata
     6781 aagccgataa caaaatcttt gtcgctcttc gcaatgtcaa cagtaccctt agtatattct
     6841 ccagtagata gggagccctt gcatgacaat tctgctaaca tcaaaaggcc tctaggttcc
     6901 tttgttactt cttctgccgc ctgcttcaaa ccgctaacaa tacctgggcc caccacaccg
     6961 tgtgcattcg taatgtctgc ccattctgct attctgtata cacccgcaga gtactgcaat
     7021 ttgactgtat taccaatgtc agcaaatttt ctgtcttcga agagtaaaaa attgtacttg
     7081 gcggataatg cctttagcgg cttaactgtg ccctccatgg aaaaatcagt caagatatcc
     7141 acatgtgttt ttagtaaaca aattttggga cctaatgctt caactaactc cagtaattcc
     7201 ttggtggtac gaacatccaa tgaagcacac aagtttgttt gcttttcgtg catgatatta
     7261 aatagcttgg cagcaacagg actaggatga gtagcagcac gttccttata tgtagctttc
     7321 gacatgattt atcttcgttt cctgcaggtt tttgttctgt gcagttgggt taagaatact
     7381 gggcaatttc atgtttcttc aacactacat atgcgtatat ataccaatct aagtctgtgc
     7441 tccttccttc gttcttcctt ctgttcggag attaccgaat caaaaaaatt tcaaagaaac
     7501 cgaaatcaaa aaaaagaata aaaaaaaaat gatgaattga attgaaaagc tgtggtatgg
     7561 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     7621 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     7681 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     7741 a
//
