LOCUS       sequence-8706-ex        7651 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 7651)
  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 7651)
  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
FEATURES             Location/Qualifiers
     source          1..7651
                     /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(5659..5678)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5660..5678)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5685..5702)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5685..5701)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5694..5716)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5846..6301
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5933..5952)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6143..6164
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6432..7235)
                     /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(7236..7456)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7482..7501)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7601..7623
                     /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 ccaatctaga ccacaaggcg gccgtggaaa ttacaaaaac ttcaactaca
     3901 ataacaattt gcaaggatat caagctggtt tccaaccaca gtctcaaggt atgtctttga
     3961 acgactttca aaagcaacaa aagcaggccg ctcccaaacc aaagaagact ttgaagcttg
     4021 tctccagttc cggtatcaag ttggccaatg ctaccaagaa ggttggcaca aaacctgccg
     4081 aatctgataa gaaagaggaa gagaagtctg ctgaaaccaa agaaccaact aaagagccaa
     4141 caaaggtcga agaaccagtt aaaaaggagg agaaaccagt ccagactgaa gaaaagacgg
     4201 aggaaaaatc ggaacttcca aaggtagaag accttaaaat ctctgaatca acacataata
     4261 ccaacaatgc caatgttacc agtgctgatg ccttgatcaa ggaacaggaa gaagaagtgg
     4321 atgacgaagt tgttaacgat ccgcggatgt ttggtggtaa agatcacgtt tctttaattt
     4381 tcatgggtca tgttgatgcc ggtaaatcta ctatgggtgg taatctacta tacttgactg
     4441 gctctgtgga taagagaact attgagaaat atgaaagaga agccaaggat gcaggcagac
     4501 aaggttggta cttgtcatgg gtcatggata ccaacaaaga agaaagaaat gatggtaaga
     4561 ctatcgaagt tggtaaggcc tactttgaaa ctgaaaaaag gcgttatacc atattggatg
     4621 ctcctggtca taaaatgtac gtttccgaga tgatcggtgg tgcttctcaa gctgatgttg
     4681 gtgttttggt catttccgcc agaaagggtg agtacgaaac cggttttgag agaggtggtc
     4741 aaactcgtga acacgcccta ttggccaaga cccaaggtgt taataagatg gttgtcgtcg
     4801 taaataagat ggatgaccca accgttaact ggtctaagga acgttacgac caatgtgtga
     4861 gtaatgtcag caatttcttg agagcaattg gttacaacat taagacagac gttgtattta
     4921 tgccagtatc cggctacagt ggtgcaaatt tgaaagatca cgtagatcca aaagaatgcc
     4981 catggtacac cggcccaact ctgttagaat atctggatac aatgaaccac gtcgaccgtc
     5041 acatcaatgc tccattcatg ttgcctattg ccgctaagat gaaggatcta ggtaccatcg
     5101 ttgaaggtaa aattgaatcc ggtcatatca aaaagggtca atccacccta ctgatgccta
     5161 acaaaaccgc tgtggaaatt caaaatattt acaacgaaac tgaaaatgaa gttgatatgg
     5221 ctatgtgtgg tgagcaagtt aaactaagaa tcaaaggtgt tgaagaagaa gacatttcac
     5281 caggttttgt actaacatcg ccaaagaacc ctatcaagag tgttaccaag tttgtagctc
     5341 aaattgctat tgtagaatta aaatctatca tagcagccgg tttttcatgt gttatgcatg
     5401 ttcatacagc aattgaagag gtacatattg ttaagttatt gcacaaatta gaaaagggta
     5461 ccaaccgtaa gtcaaagaaa ccacctgctt ttgctaagaa gggtatgaag gtcatcgctg
     5521 ttttagaaac tgaagctcca gtttgtgtgg aaacttacca agattaccct caattaggta
     5581 gattcacttt gagagatcaa ggtaccacaa tagcaattgg taaaattgtt aaaattgccg
     5641 agtaagagct ccaattcgcc ctatagtgag tcgtattaca attcactggc cgtcgtttta
     5701 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     5761 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     5821 cgcagcctga atggcgaatg gcgcgacgcg ccctgtagcg gcgcattaag cgcggcgggt
     5881 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     5941 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     6001 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     6061 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     6121 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     6181 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     6241 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     6301 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagggtaat
     6361 aactgatata attaaattga agctctaatt tgtgagttta gtatacatgc atttacttat
     6421 aatacagttt tttagttttg ctggccgcat cttctcaaat atgcttccca gcctgctttt
     6481 ctgtaacgtt caccctctac cttagcatcc cttccctttg caaatagtcc tcttccaaca
     6541 ataataatgt cagatcctgt agagaccaca tcatccacgg ttctatactg ttgacccaat
     6601 gcgtctccct tgtcatctaa acccacaccg ggtgtcataa tcaaccaatc gtaaccttca
     6661 tctcttccac ccatgtctct ttgagcaata aagccgataa caaaatcttt gtcgctcttc
     6721 gcaatgtcaa cagtaccctt agtatattct ccagtagata gggagccctt gcatgacaat
     6781 tctgctaaca tcaaaaggcc tctaggttcc tttgttactt cttctgccgc ctgcttcaaa
     6841 ccgctaacaa tacctgggcc caccacaccg tgtgcattcg taatgtctgc ccattctgct
     6901 attctgtata cacccgcaga gtactgcaat ttgactgtat taccaatgtc agcaaatttt
     6961 ctgtcttcga agagtaaaaa attgtacttg gcggataatg cctttagcgg cttaactgtg
     7021 ccctccatgg aaaaatcagt caagatatcc acatgtgttt ttagtaaaca aattttggga
     7081 cctaatgctt caactaactc cagtaattcc ttggtggtac gaacatccaa tgaagcacac
     7141 aagtttgttt gcttttcgtg catgatatta aatagcttgg cagcaacagg actaggatga
     7201 gtagcagcac gttccttata tgtagctttc gacatgattt atcttcgttt cctgcaggtt
     7261 tttgttctgt gcagttgggt taagaatact gggcaatttc atgtttcttc aacactacat
     7321 atgcgtatat ataccaatct aagtctgtgc tccttccttc gttcttcctt ctgttcggag
     7381 attaccgaat caaaaaaatt tcaaagaaac cgaaatcaaa aaaaagaata aaaaaaaaat
     7441 gatgaattga attgaaaagc tgtggtatgg tgcactctca gtacaatctg ctctgatgcc
     7501 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     7561 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     7621 aggttttcac cgtcatcacc gaaacgcgcg a
//
