LOCUS       sequence-172866-        8123 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 8123)
  AUTHORS   Liu JJ, Sondheimer N, Lindquist SL
  TITLE     Changes in the middle region of Sup35 profoundly alter the nature of
            epigenetic inheritance for the yeast prion [PSI+].
  JOURNAL   Proc Natl Acad Sci U S A 2002 Dec 10;99 Suppl 4:16446-53. Epub 2002 
            Dec 02.
  PUBMED    12461168
REFERENCE   2  (bases 1 to 8123)
  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: "Proc Natl 
            Acad Sci U S A"; date: "2002-12-10"; volume: "99 Suppl 4"; pages: 
            "16446-53. Epub 2002 Dec 02"
COMMENT     Sequence Label: pJLI-Sup35?M
FEATURES             Location/Qualifiers
     source          1..8123
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(35..52)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(206..794)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(286..305)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(965..1756,1757..1825))
                     /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     1588..1607
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1826..1930)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1998..2016
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2054..2076)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2176..2195
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2219..2434
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             2435..3238
                     /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(3369..3824)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3506..3527)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3718..3737
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3950..3972
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3964..3981
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3965..3981
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3988..4007
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3988..4006
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(4458..4481)
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     promoter        complement(7888..7906)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7927..7943)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7927..7943)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7940..7962)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7951..7967
                     /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(7975..7981,7982..7999,8000..8005))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8020..8041
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
       61 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
      121 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
      181 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
      241 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
      301 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
      361 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
      421 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
      481 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
      541 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
      601 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
      661 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
      721 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
      781 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
      841 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
      901 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
      961 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     1021 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1081 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     1141 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     1201 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     1261 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     1321 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     1381 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     1441 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     1501 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     1561 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     1621 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     1681 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     1741 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     1801 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     1861 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     1921 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     1981 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     2041 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     2101 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     2161 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat accacctttt
     2221 caattcatca tttttttttt attctttttt ttgatttcgg tttccttgaa atttttttga
     2281 ttcggtaatc tccgaacaga aggaagaacg aaggaaggag cacagactta gattggtata
     2341 tatacgcata tgtagtgttg aagaaacatg aaattgccca gtattcttaa cccaactgca
     2401 cagaacaaaa acctgcagga aacgaagata aatcatgtcg aaagctacat ataaggaacg
     2461 tgctgctact catcctagtc ctgttgctgc caagctattt aatatcatgc acgaaaagca
     2521 aacaaacttg tgtgcttcat tggatgttcg taccaccaag gaattactgg agttagttga
     2581 agcattaggt cccaaaattt gtttactaaa aacacatgtg gatatcttga ctgatttttc
     2641 catggagggc acagttaagc cgctaaaggc attatccgcc aagtacaatt ttttactctt
     2701 cgaagacaga aaatttgctg acattggtaa tacagtcaaa ttgcagtact ctgcgggtgt
     2761 atacagaata gcagaatggg cagacattac gaatgcacac ggtgtggtgg gcccaggtat
     2821 tgttagcggt ttgaagcagg cggcagaaga agtaacaaag gaacctagag gccttttgat
     2881 gttagcagaa ttgtcatgca agggctccct atctactgga gaatatacta agggtactgt
     2941 tgacattgcg aagagcgaca aagattttgt tatcggcttt attgctcaaa gagacatggg
     3001 tggaagagat gaaggttacg attggttgat tatgacaccc ggtgtgggtt tagatgacaa
     3061 gggagacgca ttgggtcaac agtatagaac cgtggatgat gtggtctcta caggatctga
     3121 cattattatt gttggaagag gactatttgc aaagggaagg gatgctaagg tagagggtga
     3181 acgttacaga aaagcaggct gggaagcata tttgagaaga tgcggccagc aaaactaaaa
     3241 aactgtatta taagtaaatg catgtatact aaactcacaa attagagctt caatttaatt
     3301 atatcagtta ttaccctatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     3361 gcatcaggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     3421 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     3481 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     3541 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     3601 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     3661 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     3721 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     3781 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc gccattcagg
     3841 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     3901 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     3961 cgttgtaaaa cgacggccag tgaattgtaa tacgactcac tatagggcga attggagctc
     4021 caccgcggtg aaaagagtca gtgagacgac gacttcagga tctttgggtt tcaggatatg
     4081 tggcatgaaa atacagaaaa atccctctgt gctgaatcag ctttcgctcg aatattacga
     4141 agaagaagca gacagtgatt atatctttat aaacaaattg tatggtcgtt caagaaccga
     4201 tcaaaatgtt tcagatgcaa ttgaacttta ttttaacaat cctcatctgt cggatgcgag
     4261 aaagcatcaa ctgaagaaaa catttttgaa aagattgcag ttgttttata atactatgct
     4321 agaagaagaa gttagaatga tatcaagtag tcttttgttt atttacgaag gagacccgga
     4381 gcgatgggaa ttactaaatg atgttgacaa acttatgcga gatgatttta tagacgatga
     4441 tgacgacgat gatgataatg atgatgatga tgatgatgat gccgagggaa gcagcgaagg
     4501 accaaaggac aaaaaaacaa ctggttcttt gagttccatg tcactaatag attttgcaca
     4561 ttctgaaata acgccgggga agggttatga tgaaaacgtg attgaaggag ttgaaacctt
     4621 gctagatatt tttatgaaat tctagatata ttgagaggtg aagtttacct tgtttatggt
     4681 atatggtaca aaaagaacta aactaattat acgtctatat atatatatat atatataaca
     4741 gctttattaa accttgtttt ttaatataga agaaaatgct ttatgatcgg tattattgtg
     4801 tttgcattta cttatgtttg caagaaatgg atccttactc ggcaatttta acaattttac
     4861 caattgctat tgtggtacct tgatctctca aagtgaatct acctaattga gggtaatctt
     4921 ggtaagtttc cacacaaact ggagcttcag tttctaaaac agcgatgacc ttcataccct
     4981 tcttagcaaa agcaggtggt ttctttgact tacggttggt acccttttct aatttgtgca
     5041 ataacttaac aatatgtacc tcttcaattg ctgtatgaac atgcataaca catgaaaaac
     5101 cggctgctat gatagatttt aattctacaa tagcaatttg agctacaaac ttggtaacac
     5161 tcttgatagg gttctttggc gatgttagta caaaacctgg tgaaatgtct tcttcttcaa
     5221 cacctttgat tcttagttta acttgctcac cacacatagc catatcaact tcattttcag
     5281 tttcgttgta aatattttga atttccacag cggttttgtt aggcatcagt agggtggatt
     5341 gacccttttt gatatgaccg gattcaattt taccttcaac gatggtacct agatccttca
     5401 tcttagcggc aataggcaac atgaatggag cattgatgtg acggtcgacg tggttcattg
     5461 tatccagata ttctaacaga gttgggccgg tgtaccatgg gcattctttt ggatctacgt
     5521 gatctttcaa atttgcacca ctgtagccgg atactggcat aaatacaacg tctgtcttaa
     5581 tgttgtaacc aattgctctc aagaaattgc tgacattact cacacattgg tcgtaacgtt
     5641 ccttagacca gttaacggtt gggtcatcca tcttatttac gacgacaacc atcttattaa
     5701 caccttgggt cttggccaat agggcgtgtt cacgagtttg accacctctc tcaaaaccgg
     5761 tttcgtactc accctttctg gcggaaatga ccaaaacacc aacatcagct tgagaagcac
     5821 caccgatcat ctcggaaacg tacattttat gaccaggagc atccaatatg gtataacgcc
     5881 ttttttcagt ttcaaagtag gccttaccaa cttcgatagt cttaccatca tttctttctt
     5941 ctttgttggt atccatgacc catgacaagt accaaccttg tctgcctgca tccttggctt
     6001 ctctttcata tttctcaata gttctcttat ccacagagcc agtcaagtat agtagattac
     6061 cacccatagt agatttaccg gcatcaacat gacccatgaa aattaaagaa acgtgatctt
     6121 taccaccaaa cataccttga gactgtggtt ggaaaccagc ttgatatcct tgcaaattgt
     6181 tattgtagtt gaagtttttg taatttccac ggccaccttg tggattgaat tgctgctgat
     6241 aaccgccttg aggattgtac tgttgatagc ctccttgagg attatactgt tgctggtaac
     6301 cggcgtcggg attgtactgt tgatagccac cttgttggta cccagaataa ccttggtaat
     6361 tttggtagta cccacctgca ggttgggctt gagcattgta agcttgataa ccttggtatc
     6421 tgttgttacc ttgttgttgg ttaccgttct ggctgtattg ctggtagttt tgctgattgt
     6481 tgccttggtt tgaatccgac atgaattctg ttgctagtgg gcagatatag atgttattcc
     6541 gagcaagtcg atgaagaaac cgctttttgt tacagtacaa tggagtcttt caagagaaga
     6601 tgtaccaata tacactacac tcttcagaag caatgggagc tttggtcgag tgaaaaaaaa
     6661 attttctcca taaagaaaga tcatattata cgatgatgta agatataata cccggttgta
     6721 atgtacattt aagagcaagg taagaagtga caataacttc tgtatgatct tagcatgtac
     6781 ctcttttggt gggctgagaa ctaagattca tctttttgcg gaagaatttt gctatgaact
     6841 tcacaacttt atgaagtggt ttaagagaat tacaaaagaa atgacacaga ctcgaacact
     6901 gtgacgcgtc gtcttagtaa aaaataataa tttgagtcaa atagcgcagc taatgcgaaa
     6961 caaagaaatg aagcatatac cattcgttgt atgatttttg tgtggttgac agatattctg
     7021 ccgaaatttt aacgcttatt ataaatataa atgtatgtat gtgtgtataa acagatacga
     7081 tattcaattt tctaccgtag ggttgggatt ttcttcaaac tccaattctt cgtcgggtat
     7141 ttcctcaatg gcgatcctct tttttggctt cggcttttca gtgtcattga caattttagg
     7201 caccttaatt tgtagtagac cgttgttgta agtagcttta atttcttcgt ccttaatgcg
     7261 tggcagcacg gggaatttaa cggttctctc aaacgcacca tattttagtt ccgtgatctt
     7321 caagaatttt tcatcaatgc ccactctgtc ttcgatctta cccttgatga gcatctcatg
     7381 agaagatgga tggtaatcaa tgtggaaagc cctagagtta gcacctggta acgcaagaac
     7441 aactacgtaa gtgtcctcgg tatcatagac attcacttct ggtgaaaatg gtaagtccat
     7501 tctcgtttca ggcttggata cttgtaacgg gtcaggtatt ggggaaggcg cttgtaggtg
     7561 agcgaacgat gaagattttt tggctaatgg tggtctcgac gattcctcca gctgattcaa
     7621 aggtttttct ttgttggttt cgccagcttc ctctttgggt gcttcagact tatccttctt
     7681 atccttttct tctccctttt cgccctcctg ttcggtattt gcttcaattt ctggttcagt
     7741 gccttcatat ggtggaacac ctattaacgc ggttaataag tcgtttaaac tgtttgcctg
     7801 ttggttggtc ctattattcc tggcagtatt acaatggtaa tatgatggat atcttctcga
     7861 gggggggccc ggtaccagct tttgttccct ttagtgaggg ttaatttcga gcttggcgta
     7921 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     7981 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     8041 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     8101 atgaatcggc caacgcgcgg gga
//
