LOCUS       sequence-8710-ex        7789 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 7789)
  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 7789)
  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+6
FEATURES             Location/Qualifiers
     source          1..7789
                     /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(5797..5816)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5798..5816)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5823..5840)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5823..5839)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5832..5854)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5984..6439
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6071..6090)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6281..6302
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6570..7373)
                     /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(7374..7594)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7620..7639)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7739..7761
                     /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 ggaggtggtt ggggtcaacc acaaggcggc cgtggaaatt
     4021 acaaaaactt caactacaat aacaatttgc aaggatatca agctggtttc caaccacagt
     4081 ctcaaggtat gtctttgaac gactttcaaa agcaacaaaa gcaggccgct cccaaaccaa
     4141 agaagacttt gaagcttgtc tccagttccg gtatcaagtt ggccaatgct accaagaagg
     4201 ttggcacaaa acctgccgaa tctgataaga aagaggaaga gaagtctgct gaaaccaaag
     4261 aaccaactaa agagccaaca aaggtcgaag aaccagttaa aaaggaggag aaaccagtcc
     4321 agactgaaga aaagacggag gaaaaatcgg aacttccaaa ggtagaagac cttaaaatct
     4381 ctgaatcaac acataatacc aacaatgcca atgttaccag tgctgatgcc ttgatcaagg
     4441 aacaggaaga agaagtggat gacgaagttg ttaacgatcc gcggatgttt ggtggtaaag
     4501 atcacgtttc tttaattttc atgggtcatg ttgatgccgg taaatctact atgggtggta
     4561 atctactata cttgactggc tctgtggata agagaactat tgagaaatat gaaagagaag
     4621 ccaaggatgc aggcagacaa ggttggtact tgtcatgggt catggatacc aacaaagaag
     4681 aaagaaatga tggtaagact atcgaagttg gtaaggccta ctttgaaact gaaaaaaggc
     4741 gttataccat attggatgct cctggtcata aaatgtacgt ttccgagatg atcggtggtg
     4801 cttctcaagc tgatgttggt gttttggtca tttccgccag aaagggtgag tacgaaaccg
     4861 gttttgagag aggtggtcaa actcgtgaac acgccctatt ggccaagacc caaggtgtta
     4921 ataagatggt tgtcgtcgta aataagatgg atgacccaac cgttaactgg tctaaggaac
     4981 gttacgacca atgtgtgagt aatgtcagca atttcttgag agcaattggt tacaacatta
     5041 agacagacgt tgtatttatg ccagtatccg gctacagtgg tgcaaatttg aaagatcacg
     5101 tagatccaaa agaatgccca tggtacaccg gcccaactct gttagaatat ctggatacaa
     5161 tgaaccacgt cgaccgtcac atcaatgctc cattcatgtt gcctattgcc gctaagatga
     5221 aggatctagg taccatcgtt gaaggtaaaa ttgaatccgg tcatatcaaa aagggtcaat
     5281 ccaccctact gatgcctaac aaaaccgctg tggaaattca aaatatttac aacgaaactg
     5341 aaaatgaagt tgatatggct atgtgtggtg agcaagttaa actaagaatc aaaggtgttg
     5401 aagaagaaga catttcacca ggttttgtac taacatcgcc aaagaaccct atcaagagtg
     5461 ttaccaagtt tgtagctcaa attgctattg tagaattaaa atctatcata gcagccggtt
     5521 tttcatgtgt tatgcatgtt catacagcaa ttgaagaggt acatattgtt aagttattgc
     5581 acaaattaga aaagggtacc aaccgtaagt caaagaaacc acctgctttt gctaagaagg
     5641 gtatgaaggt catcgctgtt ttagaaactg aagctccagt ttgtgtggaa acttaccaag
     5701 attaccctca attaggtaga ttcactttga gagatcaagg taccacaata gcaattggta
     5761 aaattgttaa aattgccgag taagagctcc aattcgccct atagtgagtc gtattacaat
     5821 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     5881 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     5941 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcgacgcgcc ctgtagcggc
     6001 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     6061 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     6121 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     6181 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     6241 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     6301 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     6361 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     6421 atattaacgt ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     6481 cacaccgcat agggtaataa ctgatataat taaattgaag ctctaatttg tgagtttagt
     6541 atacatgcat ttacttataa tacagttttt tagttttgct ggccgcatct tctcaaatat
     6601 gcttcccagc ctgcttttct gtaacgttca ccctctacct tagcatccct tccctttgca
     6661 aatagtcctc ttccaacaat aataatgtca gatcctgtag agaccacatc atccacggtt
     6721 ctatactgtt gacccaatgc gtctcccttg tcatctaaac ccacaccggg tgtcataatc
     6781 aaccaatcgt aaccttcatc tcttccaccc atgtctcttt gagcaataaa gccgataaca
     6841 aaatctttgt cgctcttcgc aatgtcaaca gtacccttag tatattctcc agtagatagg
     6901 gagcccttgc atgacaattc tgctaacatc aaaaggcctc taggttcctt tgttacttct
     6961 tctgccgcct gcttcaaacc gctaacaata cctgggccca ccacaccgtg tgcattcgta
     7021 atgtctgccc attctgctat tctgtataca cccgcagagt actgcaattt gactgtatta
     7081 ccaatgtcag caaattttct gtcttcgaag agtaaaaaat tgtacttggc ggataatgcc
     7141 tttagcggct taactgtgcc ctccatggaa aaatcagtca agatatccac atgtgttttt
     7201 agtaaacaaa ttttgggacc taatgcttca actaactcca gtaattcctt ggtggtacga
     7261 acatccaatg aagcacacaa gtttgtttgc ttttcgtgca tgatattaaa tagcttggca
     7321 gcaacaggac taggatgagt agcagcacgt tccttatatg tagctttcga catgatttat
     7381 cttcgtttcc tgcaggtttt tgttctgtgc agttgggtta agaatactgg gcaatttcat
     7441 gtttcttcaa cactacatat gcgtatatat accaatctaa gtctgtgctc cttccttcgt
     7501 tcttccttct gttcggagat taccgaatca aaaaaatttc aaagaaaccg aaatcaaaaa
     7561 aaagaataaa aaaaaaatga tgaattgaat tgaaaagctg tggtatggtg cactctcagt
     7621 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     7681 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     7741 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcga
//
