LOCUS       sequence-8712-ex        7837 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 7837)
  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 7837)
  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+8
FEATURES             Location/Qualifiers
     source          1..7837
                     /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(5845..5864)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5846..5864)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5871..5888)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5871..5887)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5880..5902)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6032..6487
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6119..6138)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6329..6350
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6618..7421)
                     /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(7422..7642)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7668..7687)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7787..7809
                     /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 atggaggagg ttggggtcaa ccccatggag
     3961 gaggttgggg tcaaccccat ggaggaggtt ggggtcaacc ccatggagga ggttggggtc
     4021 aaccccatgg aggtggttgg ggtcaaccac aaggcggccg tggaaattac aaaaacttca
     4081 actacaataa caatttgcaa ggatatcaag ctggtttcca accacagtct caaggtatgt
     4141 ctttgaacga ctttcaaaag caacaaaagc aggccgctcc caaaccaaag aagactttga
     4201 agcttgtctc cagttccggt atcaagttgg ccaatgctac caagaaggtt ggcacaaaac
     4261 ctgccgaatc tgataagaaa gaggaagaga agtctgctga aaccaaagaa ccaactaaag
     4321 agccaacaaa ggtcgaagaa ccagttaaaa aggaggagaa accagtccag actgaagaaa
     4381 agacggagga aaaatcggaa cttccaaagg tagaagacct taaaatctct gaatcaacac
     4441 ataataccaa caatgccaat gttaccagtg ctgatgcctt gatcaaggaa caggaagaag
     4501 aagtggatga cgaagttgtt aacgatccgc ggatgtttgg tggtaaagat cacgtttctt
     4561 taattttcat gggtcatgtt gatgccggta aatctactat gggtggtaat ctactatact
     4621 tgactggctc tgtggataag agaactattg agaaatatga aagagaagcc aaggatgcag
     4681 gcagacaagg ttggtacttg tcatgggtca tggataccaa caaagaagaa agaaatgatg
     4741 gtaagactat cgaagttggt aaggcctact ttgaaactga aaaaaggcgt tataccatat
     4801 tggatgctcc tggtcataaa atgtacgttt ccgagatgat cggtggtgct tctcaagctg
     4861 atgttggtgt tttggtcatt tccgccagaa agggtgagta cgaaaccggt tttgagagag
     4921 gtggtcaaac tcgtgaacac gccctattgg ccaagaccca aggtgttaat aagatggttg
     4981 tcgtcgtaaa taagatggat gacccaaccg ttaactggtc taaggaacgt tacgaccaat
     5041 gtgtgagtaa tgtcagcaat ttcttgagag caattggtta caacattaag acagacgttg
     5101 tatttatgcc agtatccggc tacagtggtg caaatttgaa agatcacgta gatccaaaag
     5161 aatgcccatg gtacaccggc ccaactctgt tagaatatct ggatacaatg aaccacgtcg
     5221 accgtcacat caatgctcca ttcatgttgc ctattgccgc taagatgaag gatctaggta
     5281 ccatcgttga aggtaaaatt gaatccggtc atatcaaaaa gggtcaatcc accctactga
     5341 tgcctaacaa aaccgctgtg gaaattcaaa atatttacaa cgaaactgaa aatgaagttg
     5401 atatggctat gtgtggtgag caagttaaac taagaatcaa aggtgttgaa gaagaagaca
     5461 tttcaccagg ttttgtacta acatcgccaa agaaccctat caagagtgtt accaagtttg
     5521 tagctcaaat tgctattgta gaattaaaat ctatcatagc agccggtttt tcatgtgtta
     5581 tgcatgttca tacagcaatt gaagaggtac atattgttaa gttattgcac aaattagaaa
     5641 agggtaccaa ccgtaagtca aagaaaccac ctgcttttgc taagaagggt atgaaggtca
     5701 tcgctgtttt agaaactgaa gctccagttt gtgtggaaac ttaccaagat taccctcaat
     5761 taggtagatt cactttgaga gatcaaggta ccacaatagc aattggtaaa attgttaaaa
     5821 ttgccgagta agagctccaa ttcgccctat agtgagtcgt attacaattc actggccgtc
     5881 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     5941 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     6001 cagttgcgca gcctgaatgg cgaatggcgc gacgcgccct gtagcggcgc attaagcgcg
     6061 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     6121 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     6181 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     6241 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     6301 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     6361 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     6421 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     6481 acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatag
     6541 ggtaataact gatataatta aattgaagct ctaatttgtg agtttagtat acatgcattt
     6601 acttataata cagtttttta gttttgctgg ccgcatcttc tcaaatatgc ttcccagcct
     6661 gcttttctgt aacgttcacc ctctacctta gcatcccttc cctttgcaaa tagtcctctt
     6721 ccaacaataa taatgtcaga tcctgtagag accacatcat ccacggttct atactgttga
     6781 cccaatgcgt ctcccttgtc atctaaaccc acaccgggtg tcataatcaa ccaatcgtaa
     6841 ccttcatctc ttccacccat gtctctttga gcaataaagc cgataacaaa atctttgtcg
     6901 ctcttcgcaa tgtcaacagt acccttagta tattctccag tagataggga gcccttgcat
     6961 gacaattctg ctaacatcaa aaggcctcta ggttcctttg ttacttcttc tgccgcctgc
     7021 ttcaaaccgc taacaatacc tgggcccacc acaccgtgtg cattcgtaat gtctgcccat
     7081 tctgctattc tgtatacacc cgcagagtac tgcaatttga ctgtattacc aatgtcagca
     7141 aattttctgt cttcgaagag taaaaaattg tacttggcgg ataatgcctt tagcggctta
     7201 actgtgccct ccatggaaaa atcagtcaag atatccacat gtgtttttag taaacaaatt
     7261 ttgggaccta atgcttcaac taactccagt aattccttgg tggtacgaac atccaatgaa
     7321 gcacacaagt ttgtttgctt ttcgtgcatg atattaaata gcttggcagc aacaggacta
     7381 ggatgagtag cagcacgttc cttatatgta gctttcgaca tgatttatct tcgtttcctg
     7441 caggtttttg ttctgtgcag ttgggttaag aatactgggc aatttcatgt ttcttcaaca
     7501 ctacatatgc gtatatatac caatctaagt ctgtgctcct tccttcgttc ttccttctgt
     7561 tcggagatta ccgaatcaaa aaaatttcaa agaaaccgaa atcaaaaaaa agaataaaaa
     7621 aaaaatgatg aattgaattg aaaagctgtg gtatggtgca ctctcagtac aatctgctct
     7681 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     7741 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     7801 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcga
//
