LOCUS       sequence-7-expor        8417 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 8417)
  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 8417)
  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 (delta)2-5
FEATURES             Location/Qualifiers
     source          1..8417
                     /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(6157..6175)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6196..6212)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6196..6212)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6209..6231)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6220..6236
                     /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(6244..6250,6251..6268,6269..6274))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(6427..6444)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6598..7186)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6678..6697)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7357..8148,8149..8217))
                     /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     7980..7999
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8218..8322)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8390..8408
                     /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 tgtgggtacc cagaataacc
     4621 ttggtaattt tggtagtacc cacctgcagg ttgggcttga gcattgtaag cttgataacc
     4681 ttggtatctg ttgttacctt gttgttggtt accgttctgg ctgtattgct ggtagttttg
     4741 ctgattgttg ccttggtttg aatccgacat gaattctgtt gctagtgggc agatatagat
     4801 gttattccga gcaagtcgat gaagaaaccg ctttttgtta cagtacaatg gagtctttca
     4861 agagaagatg taccaatata cactacactc ttcagaagca atgggagctt tggtcgagtg
     4921 aaaaaaaaat tttctccata aagaaagatc atattatacg atgatgtaag atataatacc
     4981 cggttgtaat gtacatttaa gagcaaggta agaagtgaca ataacttctg tatgatctta
     5041 gcatgtacct cttttggtgg gctgagaact aagattcatc tttttgcgga agaattttgc
     5101 tatgaacttc acaactttat gaagtggttt aagagaatta caaaagaaat gacacagact
     5161 cgaacactgt gacgcgtcgt cttagtaaaa aataataatt tgagtcaaat agcgcagcta
     5221 atgcgaaaca aagaaatgaa gcatatacca ttcgttgtat gatttttgtg tggttgacag
     5281 atattctgcc gaaattttaa cgcttattat aaatataaat gtatgtatgt gtgtataaac
     5341 agatacgata ttcaattttc taccgtaggg ttgggatttt cttcaaactc caattcttcg
     5401 tcgggtattt cctcaatggc gatcctcttt tttggcttcg gcttttcagt gtcattgaca
     5461 attttaggca ccttaatttg tagtagaccg ttgttgtaag tagctttaat ttcttcgtcc
     5521 ttaatgcgtg gcagcacggg gaatttaacg gttctctcaa acgcaccata ttttagttcc
     5581 gtgatcttca agaatttttc atcaatgccc actctgtctt cgatcttacc cttgatgagc
     5641 atctcatgag aagatggatg gtaatcaatg tggaaagccc tagagttagc acctggtaac
     5701 gcaagaacaa ctacgtaagt gtcctcggta tcatagacat tcacttctgg tgaaaatggt
     5761 aagtccattc tcgtttcagg cttggatact tgtaacgggt caggtattgg ggaaggcgct
     5821 tgtaggtgag cgaacgatga agattttttg gctaatggtg gtctcgacga ttcctccagc
     5881 tgattcaaag gtttttcttt gttggtttcg ccagcttcct ctttgggtgc ttcagactta
     5941 tccttcttat ccttttcttc tcccttttcg ccctcctgtt cggtatttgc ttcaatttct
     6001 ggttcagtgc cttcatatgg tggaacacct attaacgcgg ttaataagtc gtttaaactg
     6061 tttgcctgtt ggttggtcct attattcctg gcagtattac aatggtaata tgatggatat
     6121 cttctcgagg gggggcccgg tacccagctt ttgttccctt tagtgagggt taattccgag
     6181 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     6241 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgaggta
     6301 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     6361 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     6421 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     6481 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6541 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6601 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6661 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6721 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6781 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6841 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6901 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6961 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     7021 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     7081 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     7141 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     7201 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     7261 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     7321 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     7381 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     7441 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     7501 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7561 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7621 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7681 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7741 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7801 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7861 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7921 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7981 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     8041 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     8101 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     8161 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     8221 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     8281 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     8341 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     8401 cacgaggccc tttcgtc
//
