LOCUS       sequence-190851-        8332 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 8332)
  AUTHORS   Patino MM, Liu JJ, Glover JR, Lindquist S
  TITLE     Support for the prion hypothesis for inheritance of a phenotypic 
            trait in yeast.
  JOURNAL   Science 1996 Aug 2;273(5275):622-6.
  PUBMED    8662547
REFERENCE   2  (bases 1 to 8332)
  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: "Science"; 
            date: "1996-08-2"; volume: "273(5275)"; pages: "622-6"
COMMENT     Sequence Label: pK218TK620T
FEATURES             Location/Qualifiers
     source          1..8332
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     364..382
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(420..442)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     542..561
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        708..895
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             896..1558
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     rep_origin      complement(1818..2273)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1955..1976)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2167..2186
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2399..2421
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2413..2430
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2414..2430
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2437..2456
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2437..2455
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5905..5921)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5906..5922)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(5950..5968)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5989..6005)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5989..6005)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6002..6024)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6013..6029
                     /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(6037..6043,6044..6061,6062..6067))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6082..6103
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6220..6237)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6391..6979)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6471..6490)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7150..7941,7942..8010))
                     /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     7773..7792
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8011..8115)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    join(8145..8332,1..316)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 actttacata tatatttatt agacaagaaa agcagattaa atagatatac attcgattaa
       61 cgataagtaa aatgtaaaat cacaggattt tcgtgtgtgg tcttctacac agacaagatg
      121 aaacaattcg gcattaatac ctgagagcag gaagagcaag ataaaaggta gtatttgttg
      181 gcgatccccc tagagtcttt tacatcttcg gaaaacaaaa actatttttt ctttaatttc
      241 tttttttact ttctattttt aatttatata tttatattaa aaaatttaaa ttataattat
      301 ttttatagca cgtgatgaaa aggaccctaa gaaaccatta ttatcatgac attaacctat
      361 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
      421 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
      481 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
      541 gcggcatcag agcagattgt actgagagtg caccataaat tcccgtttta agagcttggt
      601 gagcgctagg agtcactgcc aggtatcgtt tgaacacggc attagtcagg gaagtcataa
      661 cacagtcctt tcccgcaatt ttctttttct attactcttg gcctcctcta gtacactcta
      721 tattttttta tgcctcggta atgattttca tttttttttt tccacctagc ggatgactct
      781 ttttttttct tagcgattgg cattatcaca taatgaatta tacattatat aaagtaatgt
      841 gatttcttcg aagaatatac taaaaaatga gcaggcaaga taaacgaagg caaagatgac
      901 agagcagaaa gccctagtaa agcgtattac aaatgaaacc aagattcaga ttgcgatctc
      961 tttaaagggt ggtcccctag cgatagagca ctcgatcttc ccagaaaaag aggcagaagc
     1021 agtagcagaa caggccacac aatcgcaagt gattaacgtc cacacaggta tagggtttct
     1081 ggaccatatg atacatgctc tggccaagca ttccggctgg tcgctaatcg ttgagtgcat
     1141 tggtgactta cacatagacg accatcacac cactgaagac tgcgggattg ctctcggtca
     1201 agcttttaaa gaggccctag gggccgtgcg tggagtaaaa aggtttggat caggatttgc
     1261 gcctttggat gaggcacttt ccagagcggt ggtagatctt tcgaacaggc cgtacgcagt
     1321 tgtcgaactt ggtttgcaaa gggagaaagt aggagatctc tcttgcgaga tgatcccgca
     1381 ttttcttgaa agctttgcag aggctagcag aattaccctc cacgttgatt gtctgcgagg
     1441 caagaatgat catcaccgta gtgagagtgc gttcaaggct cttgcggttg ccataagaga
     1501 agccacctcg cccaatggta ccaacgatgt tccctccacc aaaggtgttc ttatgtagtg
     1561 acaccgatta tttaaagctg cagcatacga tatatataca tgtgtatata tgtataccta
     1621 tgaatgtcag taagtatgta tacgaacagt atgatactga agatgacaag gtaatgcatc
     1681 attctatacg tgtcattctg aacgaggcgc gctttccttt tttctttttg ctttttcttt
     1741 ttttttctct tgaactcgac ggatctatgc ggtgtgaaat accgcacaga tgcgtaagga
     1801 gaaaataccg catcaggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     1861 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     1921 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     1981 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     2041 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     2101 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     2161 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     2221 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtccattcg
     2281 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     2341 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     2401 cagtcacgac gttgtaaaac gacggccagt gaattgtaat acgactcact atagggcgaa
     2461 ttggagctct tttgctcggg tgtcaagttc aattaaacaa attccattag ttttattaat
     2521 tatatatatt atattactga ttcttgttcg aaagttttta aaaatcacac tatattaaat
     2581 taatctaggt catcatcaat ttccatactg tcctcattat cgtcatcacc taacgtgtca
     2641 gcccctatag tagcttcgtg atttggtaga acttccagac attcttcagc ttcctcagga
     2701 acattttcat cacgagattt acccttcttc aagacgacat tgacagtttc cttatccttg
     2761 atttcattct ttaagatcct tagtgccagt ttgttcaaaa tttcgttttg aattaacctg
     2821 ttcagtggac gtgcacccat atcatcggaa taaccatatt tggccaagaa gtccttggcc
     2881 tcttgagtta aattcaactt gtaatgttta tcattttgct cgaatctctc ttcaatttcc
     2941 ttcaaacgaa tatccacgat cttatgaata gcttttctag atagcttgtt gaaaatgact
     3001 atactagaaa ttctgttcaa aaattctggt ctgaaatgtt gcctaacagc acccatgacc
     3061 aaattcttgg tagattcttg gatctttgat ccttgttgag aattgataaa ttcagcacct
     3121 agattggaag tcatgatgac aatacaattg gaacagtcga tcgtcttacc ttgaccagaa
     3181 gtaattctac cgtcatccaa catttgtagc atgacagtca aaacatcagg atgtgccttt
     3241 tctacttcat cgaataacaa aacggagtat ggtttgtatt gcagttggtt agttaaaaag
     3301 ccaccttcat cgtacccgac ataacctgcc gtggtaccca acaacttaga gaccgcatac
     3361 ttctcgctta attcagaaca atcgaccctg atcatcatgt cctcatcatt aaacaaaaat
     3421 ccagcaactt ttttagccaa ttcagtggta ccggaaccgg acaaacctaa aaataagaag
     3481 gatgctggtt gccttggatt agctaaacct gatctagaca atctaacggc attggaaaca
     3541 gctttaatgg catccatttg gcccacgact tcagatgata agtcacgttc catatgaatc
     3601 aatttttcat tttcagattc tgacaacttc ttaacaggga taccagtcaa tcttgcagct
     3661 gtttcagaaa tggtgtctga atcgaccaca ttttggatca tggagttggc accagcacgt
     3721 ctctcttcct cagcaacctg atcttcaagc ttttcgattt gctttttgat atctgggatg
     3781 gcgaagtacc ttaaatcagc ggcggtagca gtatcatatc tacgttcagc atcaagggcc
     3841 ttgttttcca gttcatccaa tttcttttta gcttgtgtca attcttcatg gccatgcttt
     3901 tcttcattgt aacgttgtct tagaggttcc aattcttctt gcaatgaagc ttccttctgc
     3961 ctagctaact ttaatctatc tttagtggtg gagtcggcat cttcatctct ctctagagct
     4021 tttatctcta cttgaatcaa ttgcaattga cgttccttgg aatccaattc ttctggctta
     4081 gaatctcttg cgacggcgac accagcacaa gaaatatcaa ctaaatccaa agcagaatct
     4141 ggcaatcttc tatatggcaa gtaacgcttg gctaattgag cagcagtgac taaggcgcta
     4201 tccagaatcc ttacaccatg atgtatttca tactttggtt gcagacctct caatatggcc
     4261 actgtttgtc tcacacttgg ttcagcgact tcaattttct ggaatcttct ttcaaaggca
     4321 ccatcctttt ccacaataga tctatattcg ttattggtgg tggcaccgat gaccttcaat
     4381 tggcctctgg acaaagctgg cttcaagatg ttagcagcgt cgtccttacc attacccatt
     4441 aacatgtgaa tttcatcaat gaataacaca attagagtct ttgattcttc gatttccttc
     4501 aaaacacctt tgaatctttc ttcgaaatca cctttgtatt tagcacctgc ggttaatgcg
     4561 gccaaatcta gactgaacaa tttagcgcct tgtaagatag tgggaacgtc atcgtcaatg
     4621 attctttgag caacaccttc aataatagcg gtggtaccga tacctggctc accaattaaa
     4681 catgggttgg acttaattct tcttgctaaa actctaatag tgcttcttat ttcttcttca
     4741 cggccgatga cagggtcaag tttaccttga cgagcctgct cagtcatatc aatggcgtac
     4801 tttgataaat attccaaagg tgtgttcgta tcagcgccac gagagtcaat tctagtgtta
     4861 ccacgaagtt caagagcttg ttgcttgatg gcctcaatat ctacttgagc ttccttaaat
     4921 atttgctgaa tagacgaatc attgaataga gcaaacaata tatggtcttg cgctataaat
     4981 gagtccttct gttgtttttg aatcttagca gcgtcttgaa ggactttccc caaagcataa
     5041 cttggagtta tctccgcagg tgcaggttgc tgttgaggaa ttcttactag atttctatta
     5101 accactttct tgaaaagatc atagtcgtaa cggcccttct caattagatt ctgtaggtaa
     5161 gggactgatc catcttctgg cgtttcaatg aaggcagcta gaatatgtat aggttgtaat
     5221 tgtggatgtt gatgatccga agccaatttt tgagccaacg tcaaaatcgt tagagccctt
     5281 tctgtaaatt gcgtttggtc gttcatatat tctgtatatt ttatggtacg tgtagttgat
     5341 ttcttttttc tttgttattg ctgattcgat tcaagggtaa aaaggaacaa gtgacaaagg
     5401 aacgaattat gaaggaattg aggcaagatt acaatgcctt atataataat ctggcaggtt
     5461 gcataagttt gcgccccttt gccttttact gtgatataat ttgcagttct ttgagatggg
     5521 ccccctgttg aaaattgaac aaaaaaatcg cggcccctta tggaacgttt aagattgtaa
     5581 ttaataaccc ttctagaaaa ttctggaaaa atatatatct acaagaacaa cgaaagaaaa
     5641 gttaggaaca gcaaacacta ccacgatgga gggttcaatg ttaatatgta tgataaatat
     5701 gtaaataaat aagtaaatga ataaataagt gaataggtag attctttttt tctaaaaatg
     5761 gtgcagaagg tttttaaacc atgaaactct cgccacaacc gcatgtgccc ttggagtttg
     5821 gattcttgaa gacaaactta gaggccaact tgtcgtcgac ccagtccatt tcacttccaa
     5881 tgatgctgaa taacgccttt gaatcgatac cgtcgacctc gagggggggc ccggtaccag
     5941 cttttgttcc ctttagtgag ggttaatttc gagcttggcg taatcatggt catagctgtt
     6001 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6061 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6121 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6181 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6241 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6301 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6361 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6421 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6481 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6541 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6601 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6661 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6721 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6781 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     6841 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6901 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6961 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     7021 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7081 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7141 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7201 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7261 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7321 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7381 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7441 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7501 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7561 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7621 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7681 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7741 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7801 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7861 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7921 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7981 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     8041 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8101 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacggatcgc ttgcctgtaa
     8161 cttacacgcg cctcgtatct tttaatgatg gaataatttg ggaatttact ctgtgtttat
     8221 ttatttttat gttttgtatt tggattttag aaagtaaata aagaaggtag aagagttacg
     8281 gaatgaagaa aaaaaaataa acaaaggttt aaaaaatttc aacaaaaagc gt
//
