LOCUS       sequence-343546-        6467 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 6467)
  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 6467)
  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
FEATURES             Location/Qualifiers
     source          1..6467
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     47..65
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(103..125)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     225..244
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        268..483
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             484..1287
                     /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(1418..1873)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1555..1576)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1767..1786
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1999..2021
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2013..2030
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2014..2030
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2037..2056
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2037..2055
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(2507..2530)
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     promoter        complement(4281..4299)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4320..4336)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4320..4336)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4333..4355)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4344..4360
                     /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(4368..4374,4375..4392,4393..4398))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4413..4434
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4551..4568)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4722..5310)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4802..4821)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5481..6272,6273..6341))
                     /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     6104..6123
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6342..6446)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
       61 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
      121 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
      181 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
      241 tgtactgaga gtgcaccata ccaccttttc aattcatcat ttttttttta ttcttttttt
      301 tgatttcggt ttccttgaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga
      361 aggaaggagc acagacttag attggtatat atacgcatat gtagtgttga agaaacatga
      421 aattgcccag tattcttaac ccaactgcac agaacaaaaa cctgcaggaa acgaagataa
      481 atcatgtcga aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc
      541 aagctattta atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt
      601 accaccaagg aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa
      661 acacatgtgg atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca
      721 ttatccgcca agtacaattt tttactcttc gaagacagaa aatttgctga cattggtaat
      781 acagtcaaat tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg
      841 aatgcacacg gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa
      901 gtaacaaagg aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta
      961 tctactggag aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt
     1021 atcggcttta ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt
     1081 atgacacccg gtgtgggttt agatgacaag ggagacgcat tgggtcaaca gtatagaacc
     1141 gtggatgatg tggtctctac aggatctgac attattattg ttggaagagg actatttgca
     1201 aagggaaggg atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat
     1261 ttgagaagat gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta
     1321 aactcacaaa ttagagcttc aatttaatta tatcagttat taccctatgc ggtgtgaaat
     1381 accgcacaga tgcgtaagga gaaaataccg catcaggaaa ttgtaagcgt taatattttg
     1441 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     1501 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     1561 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     1621 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     1681 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     1741 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     1801 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     1861 ctacagggcg cgtccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     1921 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     1981 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattgtaat
     2041 acgactcact atagggcgaa ttggagctcc accgcggtga aaagagtcag tgagacgacg
     2101 acttcaggat ctttgggttt caggatatgt ggcatgaaaa tacagaaaaa tccctctgtg
     2161 ctgaatcagc tttcgctcga atattacgaa gaagaagcag acagtgatta tatctttata
     2221 aacaaattgt atggtcgttc aagaaccgat caaaatgttt cagatgcaat tgaactttat
     2281 tttaacaatc ctcatctgtc ggatgcgaga aagcatcaac tgaagaaaac atttttgaaa
     2341 agattgcagt tgttttataa tactatgcta gaagaagaag ttagaatgat atcaagtagt
     2401 cttttgttta tttacgaagg agacccggag cgatgggaat tactaaatga tgttgacaaa
     2461 cttatgcgag atgattttat agacgatgat gacgacgatg atgataatga tgatgatgat
     2521 gatgatgatg ccgagggaag cagcgaagga ccaaaggaca aaaaaacaac tggttctttg
     2581 agttccatgt cactaataga ttttgcacat tctgaaataa cgccggggaa gggttatgat
     2641 gaaaacgtga ttgaaggagt tgaaaccttg ctagatattt ttatgaaatt ctagatatat
     2701 tgagaggtga agtttacctt gtttatggta tatggtacaa aaagaactaa actaattata
     2761 cgtctatata tatatatata tatataacag ctttattaaa ccttgttttt taatatagaa
     2821 gaaaatgctt tatgatcggt attattgtgt ttgcatttac ttatgtttgc aagaaatgga
     2881 tcccccgggc tgcaggaatt ctgttgctag tgggcagata tagatgttat tccgagcaag
     2941 tcgatgaaga aaccgctttt tgttacagta caatggagtc tttcaagaga agatgtacca
     3001 atatacacta cactcttcag aagcaatggg agctttggtc gagtgaaaaa aaaattttct
     3061 ccataaagaa agatcatatt atacgatgat gtaagatata atacccggtt gtaatgtaca
     3121 tttaagagca aggtaagaag tgacaataac ttctgtatga tcttagcatg tacctctttt
     3181 ggtgggctga gaactaagat tcatcttttt gcggaagaat tttgctatga acttcacaac
     3241 tttatgaagt ggtttaagag aattacaaaa gaaatgacac agactcgaac actgtgacgc
     3301 gtcgtcttag taaaaaataa taatttgagt caaatagcgc agctaatgcg aaacaaagaa
     3361 atgaagcata taccattcgt tgtatgattt ttgtgtggtt gacagatatt ctgccgaaat
     3421 tttaacgctt attataaata taaatgtatg tatgtgtgta taaacagata cgatattcaa
     3481 ttttctaccg tagggttggg attttcttca aactccaatt cttcgtcggg tatttcctca
     3541 atggcgatcc tcttttttgg cttcggcttt tcagtgtcat tgacaatttt aggcacctta
     3601 atttgtagta gaccgttgtt gtaagtagct ttaatttctt cgtccttaat gcgtggcagc
     3661 acggggaatt taacggttct ctcaaacgca ccatatttta gttccgtgat cttcaagaat
     3721 ttttcatcaa tgcccactct gtcttcgatc ttacccttga tgagcatctc atgagaagat
     3781 ggatggtaat caatgtggaa agccctagag ttagcacctg gtaacgcaag aacaactacg
     3841 taagtgtcct cggtatcata gacattcact tctggtgaaa atggtaagtc cattctcgtt
     3901 tcaggcttgg atacttgtaa cgggtcaggt attggggaag gcgcttgtag gtgagcgaac
     3961 gatgaagatt ttttggctaa tggtggtctc gacgattcct ccagctgatt caaaggtttt
     4021 tctttgttgg tttcgccagc ttcctctttg ggtgcttcag acttatcctt cttatccttt
     4081 tcttctccct tttcgccctc ctgttcggta tttgcttcaa tttctggttc agtgccttca
     4141 tatggtggaa cacctattaa cgcggttaat aagtcgttta aactgtttgc ctgttggttg
     4201 gtcctattat tcctggcagt attacaatgg taatatgatg gatatcttct cgaggggggg
     4261 cccggtacca gcttttgttc cctttagtga gggttaattt cgagcttggc gtaatcatgg
     4321 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4381 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     4441 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4501 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4561 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4621 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4681 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4741 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4801 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4861 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4921 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4981 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5041 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5101 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5161 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5221 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5281 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5341 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5401 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5461 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5521 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5581 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5641 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5701 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5761 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5821 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5881 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5941 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6001 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6061 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6121 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6181 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6241 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6301 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6361 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6421 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgcc
//
