LOCUS       sequence-314743-        8314 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 8314)
  AUTHORS   Glover JR, Kowal AS, Schirmer EC, Patino MM, Liu JJ, Lindquist S
  TITLE     Self-seeded fibers formed by Sup35, the protein determinant of 
            [PSI+], a heritable prion-like factor of S. cerevisiae.
  JOURNAL   Cell 1997 May 30;89(5):811-9.
  PUBMED    9182769
REFERENCE   2  (bases 1 to 8314)
  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: "Cell"; 
            date: "1997-05-30"; volume: "89(5)"; pages: "811-9"
COMMENT     Sequence Label: pRSCupSup35GFP
FEATURES             Location/Qualifiers
     source          1..8314
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     1058..1074
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1059..1075
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1496..1730
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     CDS             3845..4567
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYKG"
     primer_bind     complement(3878..3906)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4501..4521
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4589..4608)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4590..4608)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4615..4632)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4615..4631)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4624..4646)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4772..5227
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4859..4878)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5069..5090
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5358..6161)
                     /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(6162..6377)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6401..6420)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6520..6542
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6580..6598)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    6639..7142
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        7179..7283
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7284..7352,7353..8144)
                     /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(7502..7521)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctcg aaattaaccc
     1021 tcactaaagg gaacaaaagc tggtaccggg ccccccctcg aggtcgacgg tatcgataag
     1081 cttgatatcg aatttcccat taccgacatt tgggcggcgg ccaaagcggt cggacagtgc
     1141 tccgagaacg ggtgcgcata gaaattgcat caacgcatat agcgctagca gcacgccata
     1201 gtgactggcg atgctgtcgg aatggacgat atcccgcaag aggcccggca gtaccggcat
     1261 aaccaagcct atgcctacag catccagggt gacggtgccg aggatgacga tgagcgcatt
     1321 gttagatttc atacacggtg cctgactgcg ttagcaattt aactgtgata aactaccgca
     1381 ttaaagctaa aacacttttg tattattttt cctcatatat gtgtataggt ttatacggat
     1441 gatttaatta ttacttcacc accctttatt tcaggctgat atcttagcct tgttactagt
     1501 tagaaaaaga catttttgct gtcagtcact gtcaagagat tcttttgctg gcatttcttc
     1561 tagaagcaaa aagagcgatg cgtcttttcc gctgaaccgt tccagcaaaa aagactacca
     1621 acgcaatatg gattgtcaga atcatataaa agagaagcaa ataactcctt gtcttgtatc
     1681 aattgcatta taatatcttc ttgttagtgc aatatcatat agaagtcatc gaaatagata
     1741 ttaagaaaaa caaactgtac aatcaatcaa tcaatcagga tccatgtcgg attcaaacca
     1801 aggcaacaat cagcaaaact accagcaata cagccagaac ggtaaccaac aacaaggtaa
     1861 caacagatac caaggttatc aagcttacaa tgctcaagcc caacctgcag gtgggtacta
     1921 ccaaaattac caaggttatt ctgggtacca acaaggtggc tatcaacagt acaatcccga
     1981 cgccggttac cagcaacagt ataatcctca aggaggctat caacagtaca atcctcaagg
     2041 cggttatcag cagcaattca atccacaagg tggccgtgga aattacaaaa acttcaacta
     2101 caataacaat ttgcaaggat atcaagctgg tttccaacca cagtctcaag gtatgtcttt
     2161 gaacgacttt caaaagcaac aaaagcaggc cgctcccaaa ccaaagaaga ctttgaagct
     2221 tgtctccagt tccggtatca agttggccaa tgctaccaag aaggttggca caaaacctgc
     2281 cgaatctgat aagaaagagg aagagaagtc tgctgaaacc aaagaaccaa ctaaagagcc
     2341 aacaaaggtc gaagaaccag ttaaaaagga ggagaaacca gtccagactg aagaaaagac
     2401 ggaggaaaaa tcggaacttc caaaggtaga agaccttaaa atctctgaat caacacataa
     2461 taccaacaat gccaatgtta ccagtgctga tgccttgatc aaggaacagg aagaagaagt
     2521 ggatgacgaa gttgttaacg atatgtttgg tggtaaagat cacgtttctt taattttcat
     2581 gggtcatgtt gatgccggta aatctactat gggtggtaat ctactatact tgactggctc
     2641 tgtggataag agaactattg agaaatatga aagagaagcc aaggatgcag gcagacaagg
     2701 ttggtacttg tcatgggtca tggataccaa caaagaagaa agaaatgatg gtaagactat
     2761 cgaagttggt aaggcctact ttgaaactga aaaaaggcgt tataccatat tggatgctcc
     2821 tggtcataaa atgtacgttt ccgagatgat cggtggtgct tctcaagctg atgttggtgt
     2881 tttggtcatt tccgccagaa agggtgagta cgaaaccggt tttgagagag gtggtcaaac
     2941 tcgtgaacac gccctattgg ccaagaccca aggtgttaat aagatggttg tcgtcgtaaa
     3001 taagatggat gacccaaccg ttaactggtc taaggaacgt tacgaccaat gtgtgagtaa
     3061 tgtcagcaat ttcttgagag caattggtta caacattaag acagacgttg tatttatgcc
     3121 agtatccggc tacagtggtg caaatttgaa agatcacgta gatccaaaag aatgcccatg
     3181 gtacaccggc ccaactctgt tagaatatct ggatacaatg aaccacgtcg accgtcacat
     3241 caatgctcca ttcatgttgc ctattgccgc taagatgaag gatctaggta ccatcgttga
     3301 aggtaaaatt gaatccggtc atatcaaaaa gggtcaatcc accctactga tgcctaacaa
     3361 aaccgctgtg gaaattcaaa atatttacaa cgaaactgaa aatgaagttg atatggctat
     3421 gtgtggtgag caagttaaac taagaatcaa aggtgttgaa gaagaagaca tttcaccagg
     3481 ttttgtacta acatcgccaa agaaccctat caagagtgtt accaagtttg tagctcaaat
     3541 tgctattgta gaattaaaat ctatcatagc agccggtttt tcatgtgtta tgcatgttca
     3601 tacagcaatt gaagaggtac atattgttaa gttattgcac aaattagaaa agggtaccaa
     3661 ccgtaagtca aagaaaccac ctgcttttgc taagaagggt atgaaggtca tcgctgtttt
     3721 agaaactgaa gctccagttt gtgtggaaac ttaccaagat taccctcaat taggtagatt
     3781 cactttgaga gatcaaggta ccacaatagc aattggtaaa attgttaaaa ttgccgagga
     3841 gctcatggct agcaaaggag aagaactctt cactggagtt gtcccaattc ttgttgaatt
     3901 agatggtgat gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgcaac
     3961 atacggaaaa cttaccctta aatttatttg cactactgga aaactacctg ttccatggcc
     4021 aacacttgtc actactttca cttatggtgt tcagtgcttt tcaagatacc cggatcatat
     4081 gaaacggcat gactttttca agagtgccat gcccgaaggt tatgtacagg aaagaactat
     4141 atttttcaaa gatgacggga actacaagac acgtgctgaa gtcaagtttg aaggtgatac
     4201 ccttgttaat agaatcgagt taaaaggtat tgattttaaa gaagatggaa acattcttgg
     4261 gcacaaattg gaatacaact ataactcaca caatgtatac atcatggcag acaaacaaaa
     4321 gaatggaatc aaagctaact tcaaaattag acacaacatt gaagatggaa gcgttcaact
     4381 agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa
     4441 ccattacctg tccacacaat ctgccctttc gaaagatccc aacgaaaaga gagaccacat
     4501 ggtccttctt gagtttgtaa cagctgctgg gattacacat ggcatggatg aactatacaa
     4561 aggatgaccg cggtggagct ccaattcgcc ctatagtgag tcgtattaca attcactggc
     4621 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     4681 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     4741 ccaacagttg cgcagcctga atggcgaatg gacgcgccct gtagcggcgc attaagcgcg
     4801 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4861 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4921 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4981 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     5041 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     5101 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     5161 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     5221 acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatag
     5281 ggtaataact gatataatta aattgaagct ctaatttgtg agtttagtat acatgcattt
     5341 acttataata cagtttttta gttttgctgg ccgcatcttc tcaaatatgc ttcccagcct
     5401 gcttttctgt aacgttcacc ctctacctta gcatcccttc cctttgcaaa tagtcctctt
     5461 ccaacaataa taatgtcaga tcctgtagag accacatcat ccacggttct atactgttga
     5521 cccaatgcgt ctcccttgtc atctaaaccc acaccgggtg tcataatcaa ccaatcgtaa
     5581 ccttcatctc ttccacccat gtctctttga gcaataaagc cgataacaaa atctttgtcg
     5641 ctcttcgcaa tgtcaacagt acccttagta tattctccag tagataggga gcccttgcat
     5701 gacaattctg ctaacatcaa aaggcctcta ggttcctttg ttacttcttc tgccgcctgc
     5761 ttcaaaccgc taacaatacc tgggcccacc acaccgtgtg cattcgtaat gtctgcccat
     5821 tctgctattc tgtatacacc cgcagagtac tgcaatttga ctgtattacc aatgtcagca
     5881 aattttctgt cttcgaagag taaaaaattg tacttggcgg ataatgcctt tagcggctta
     5941 actgtgccct ccatggaaaa atcagtcaag atatccacat gtgtttttag taaacaaatt
     6001 ttgggaccta atgcttcaac taactccagt aattccttgg tggtacgaac atccaatgaa
     6061 gcacacaagt ttgtttgctt ttcgtgcatg atattaaata gcttggcagc aacaggacta
     6121 ggatgagtag cagcacgttc cttatatgta gctttcgaca tgatttatct tcgtttcctg
     6181 caggtttttg ttctgtgcag ttgggttaag aatactgggc aatttcatgt ttcttcaaca
     6241 ctacatatgc gtatatatac caatctaagt ctgtgctcct tccttcgttc ttccttctgt
     6301 tcggagatta ccgaatcaaa aaaatttcaa ggaaaccgaa atcaaaaaaa agaataaaaa
     6361 aaaaatgatg aattgaaaag gtggtatggt gcactctcag tacaatctgc tctgatgccg
     6421 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     6481 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     6541 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     6601 tataggttaa tgtcatgata ataatggttt cttagacgga tcgcttgcct gtaacttaca
     6661 cgcgcctcgt atcttttaat gatggaataa tttgggaatt tactctgtgt ttatttattt
     6721 ttatgttttg tatttggatt ttagaaagta aataaagaag gtagaagagt tacggaatga
     6781 agaaaaaaaa ataaacaaag gtttaaaaaa tttcaacaaa aagcgtactt tacatatata
     6841 tttattagac aagaaaagca gattaaatag atatacattc gattaacgat aagtaaaatg
     6901 taaaatcaca ggattttcgt gtgtggtctt ctacacagac aagatgaaac aattcggcat
     6961 taatacctga gagcaggaag agcaagataa aaggtagtat ttgttggcga tccccctaga
     7021 gtcttttaca tcttcggaaa acaaaaacta ttttttcttt aatttctttt tttactttct
     7081 atttttaatt tatatattta tattaaaaaa tttaaattat aattattttt atagcacgtg
     7141 atgaaaagga cccaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     7201 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     7261 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     7321 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     7381 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     7441 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     7501 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     7561 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     7621 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     7681 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     7741 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     7801 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     7861 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     7921 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     7981 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     8041 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     8101 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     8161 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     8221 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     8281 cgtcagaccc cgtagaaaag atcaaaggat cttc
//
