LOCUS       sequence-6-expor        8585 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 8585)
  AUTHORS   Liu JJ, Lindquist S
  TITLE     Oligopeptide-repeat expansions modulate 'protein-only' inheritance 
            in yeast.
  JOURNAL   Nature 1999 Aug 5;400(6744):573-6.
  PUBMED    10448860
REFERENCE   2  (bases 1 to 8585)
  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: "Nature"; 
            date: "1999-08-5"; volume: "400(6744)"; pages: "573-6"
COMMENT     Sequence Label: pJLI-SUP35 R2E2
FEATURES             Location/Qualifiers
     source          1..8585
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        196..416
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             417..1220
                     /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"
     rep_origin      complement(1351..1806)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1488..1509)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1700..1719
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1936..1958
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1950..1967
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1951..1967
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1974..1993
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1974..1992
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(2444..2467)
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     promoter        complement(6325..6343)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6364..6380)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6364..6380)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6377..6399)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6388..6404
                     /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)."
     promoter        complement(join(6412..6418,6419..6436,6437..6442))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(6595..6612)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6766..7354)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6846..6865)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7525..8316,8317..8385))
                     /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     8148..8167
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8386..8490)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8558..8576
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac agcttttcaa ttcaattcat catttttttt ttattctttt ttttgatttc
      241 ggtttctttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
      301 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc
      361 cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaatcatgt
      421 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat
      481 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca
      541 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg
      601 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
      661 ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt aatacagtca
      721 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
      781 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
      841 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
      901 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
      961 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
     1021 ccggtgtggg tttagatgac aagggagacg cattgggtca acagtataga accgtggatg
     1081 atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
     1141 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
     1201 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac
     1261 aaattagagc ttcaatttaa ttatatcagt tattacccta tgcggtgtga aataccgcac
     1321 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat
     1381 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     1441 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     1501 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     1561 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     1621 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     1681 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     1741 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     1801 gcgcgtcgcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     1861 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     1921 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttgtaatacg
     1981 actcactata gggcgaattg gagctccacc gcggtgaaaa gagtcagtga gacgacgact
     2041 tcaggatctt tgggtttcag gatatgtggc atgaaaatac agaaaaatcc ctctgtgctg
     2101 aatcagcttt cgctcgaata ttacgaagaa gaagcagaca gtgattatat ctttataaac
     2161 aaattgtatg gtcgttcaag aaccgatcaa aatgtttcag atgcaattga actttatttt
     2221 aacaatcctc atctgtcgga tgcgagaaag catcaactga agaaaacatt tttgaaaaga
     2281 ttgcagttgt tttataatac tatgctagaa gaagaagtta gaatgatatc aagtagtctt
     2341 ttgtttattt acgaaggaga cccggagcga tgggaattac taaatgatgt tgacaaactt
     2401 atgcgagatg attttataga cgatgatgac gacgatgatg ataatgatga tgatgatgat
     2461 gatgatgccg agggaagcag cgaaggacca aaggacaaaa aaacaactgg ttctttgagt
     2521 tccatgtcac taatagattt tgcacattct gaaataacgc cggggaaggg ttatgatgaa
     2581 aacgtgattg aaggagttga aaccttgcta gatattttta tgaaattcta gatatattga
     2641 gaggtgaagt ttaccttgtt tatggtatat ggtacaaaaa gaactaaact aattatacgt
     2701 ctatatatat atatatatat ataacagctt tattaaacct tgttttttaa tatagaagaa
     2761 aatgctttat gatcggtatt attgtgtttg catttactta tgtttgcaag aaatggatcc
     2821 ttactcggca attttaacaa ttttaccaat tgctattgtg gtaccttgat ctctcaaagt
     2881 gaatctacct aattgagggt aatcttggta agtttccaca caaactggag cttcagtttc
     2941 taaaacagcg atgaccttca tacccttctt agcaaaagca ggtggtttct ttgacttacg
     3001 gttggtaccc ttttctaatt tgtgcaataa cttaacaata tgtacctctt caattgctgt
     3061 atgaacatgc ataacacatg aaaaaccggc tgctatgata gattttaatt ctacaatagc
     3121 aatttgagct acaaacttgg taacactctt gatagggttc tttggcgatg ttagtacaaa
     3181 acctggtgaa atgtcttctt cttcaacacc tttgattctt agtttaactt gctcaccaca
     3241 catagccata tcaacttcat tttcagtttc gttgtaaata ttttgaattt ccacagcggt
     3301 tttgttaggc atcagtaggg tggattgacc ctttttgata tgaccggatt caattttacc
     3361 ttcaacgatg gtacctagat ccttcatctt agcggcaata ggcaacatga atggagcatt
     3421 gatgtgacgg tcgacgtggt tcattgtatc cagatattct aacagagttg ggccggtgta
     3481 ccatgggcat tcttttggat ctacgtgatc tttcaaattt gcaccactgt agccggatac
     3541 tggcataaat acaacgtctg tcttaatgtt gtaaccaatt gctctcaaga aattgctgac
     3601 attactcaca cattggtcgt aacgttcctt agaccagtta acggttgggt catccatctt
     3661 atttacgacg acaaccatct tattaacacc ttgggtcttg gccaataggg cgtgttcacg
     3721 agtttgacca cctctctcaa aaccggtttc gtactcaccc tttctggcgg aaatgaccaa
     3781 aacaccaaca tcagcttgag aagcaccacc gatcatctcg gaaacgtaca ttttatgacc
     3841 aggagcatcc aatatggtat aacgcctttt ttcagtttca aagtaggcct taccaacttc
     3901 gatagtctta ccatcatttc tttcttcttt gttggtatcc atgacccatg acaagtacca
     3961 accttgtctg cctgcatcct tggcttctct ttcatatttc tcaatagttc tcttatccac
     4021 agagccagtc aagtatagta gattaccacc catagtagat ttaccggcat caacatgacc
     4081 catgaaaatt aaagaaacgt gatctttacc accaaacata tcgttaacaa cttcgtcatc
     4141 cacttcttct tcctgttcct tgatcaaggc atcagcactg gtaacattgg cattgttggt
     4201 attatgtgtt gattcagaga ttttaaggtc ttctaccttt ggaagttccg atttttcctc
     4261 cgtcttttct tcagtctgga ctggtttctc ctccttttta actggttctt cgacctttgt
     4321 tggctcttta gttggttctt tggtttcagc agacttctct tcctctttct tatcagattc
     4381 ggcaggtttt gtgccaacct tcttggtagc attggccaac ttgataccgg aactggagac
     4441 aagcttcaaa gtcttctttg gtttgggagc ggcctgcttt tgttgctttt gaaagtcgtt
     4501 caaagacatg aattcacctt gagactgtgg ttggaaacca gcttgatatc cttgcaaatt
     4561 gttattgtag ttgaagtttt tgtaatttcc acggccacct tgtggattga attgctgctg
     4621 ataaccgcct tgaggattgt actgttgata gcctccttga ggattatact gttgctggta
     4681 accggcgtcg ggattgtact gttgatagcc accttgggga ttgtactgtt gatagccacc
     4741 ttggggattg tactgttgat agccaccttg ttggtaccca gaataacctt ggtaattttg
     4801 gtagtaccca cctgcaggtt gggcttgagc attgtaagct tgataacctt ggtatctgtt
     4861 gttaccttgt tgttggttac cgttctggct gtattgctgg tagttttgct gattgttgcc
     4921 ttggtttgaa tccgacatga attctgttgc tagtgggcag atatagatgt tattccgagc
     4981 aagtcgatga agaaaccgct ttttgttaca gtacaatgga gtctttcaag agaagatgta
     5041 ccaatataca ctacactctt cagaagcaat gggagctttg gtcgagtgaa aaaaaaattt
     5101 tctccataaa gaaagatcat attatacgat gatgtaagat ataatacccg gttgtaatgt
     5161 acatttaaga gcaaggtaag aagtgacaat aacttctgta tgatcttagc atgtacctct
     5221 tttggtgggc tgagaactaa gattcatctt tttgcggaag aattttgcta tgaacttcac
     5281 aactttatga agtggtttaa gagaattaca aaagaaatga cacagactcg aacactgtga
     5341 cgcgtcgtct tagtaaaaaa taataatttg agtcaaatag cgcagctaat gcgaaacaaa
     5401 gaaatgaagc atataccatt cgttgtatga tttttgtgtg gttgacagat attctgccga
     5461 aattttaacg cttattataa atataaatgt atgtatgtgt gtataaacag atacgatatt
     5521 caattttcta ccgtagggtt gggattttct tcaaactcca attcttcgtc gggtatttcc
     5581 tcaatggcga tcctcttttt tggcttcggc ttttcagtgt cattgacaat tttaggcacc
     5641 ttaatttgta gtagaccgtt gttgtaagta gctttaattt cttcgtcctt aatgcgtggc
     5701 agcacgggga atttaacggt tctctcaaac gcaccatatt ttagttccgt gatcttcaag
     5761 aatttttcat caatgcccac tctgtcttcg atcttaccct tgatgagcat ctcatgagaa
     5821 gatggatggt aatcaatgtg gaaagcccta gagttagcac ctggtaacgc aagaacaact
     5881 acgtaagtgt cctcggtatc atagacattc acttctggtg aaaatggtaa gtccattctc
     5941 gtttcaggct tggatacttg taacgggtca ggtattgggg aaggcgcttg taggtgagcg
     6001 aacgatgaag attttttggc taatggtggt ctcgacgatt cctccagctg attcaaaggt
     6061 ttttctttgt tggtttcgcc agcttcctct ttgggtgctt cagacttatc cttcttatcc
     6121 ttttcttctc ccttttcgcc ctcctgttcg gtatttgctt caatttctgg ttcagtgcct
     6181 tcatatggtg gaacacctat taacgcggtt aataagtcgt ttaaactgtt tgcctgttgg
     6241 ttggtcctat tattcctggc agtattacaa tggtaatatg atggatatct tctcgagggg
     6301 gggcccggta cccagctttt gttcccttta gtgagggtta attccgagct tggcgtaatc
     6361 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     6421 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgaggtaac tcacattaat
     6481 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     6541 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     6601 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     6661 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     6721 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     6781 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6841 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6901 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6961 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     7021 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     7081 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     7141 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     7201 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     7261 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     7321 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     7381 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     7441 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     7501 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     7561 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     7621 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     7681 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     7741 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     7801 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     7861 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7921 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     7981 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     8041 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     8101 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     8161 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     8221 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     8281 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     8341 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     8401 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     8461 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     8521 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     8581 tcgtc
//
