LOCUS       sequence-3-expor        8451 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 8451)
  AUTHORS   Li L, Lindquist S
  TITLE     Creating a protein-based element of inheritance.
  JOURNAL   Science 2000 Jan 28;287(5453):661-4.
  PUBMED    10650001
REFERENCE   2  (bases 1 to 8451)
  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: "2000-01-28"; volume: "287(5453)"; pages: "661-4"
COMMENT     Sequence Label: pCUP1-NMGR
FEATURES             Location/Qualifiers
     source          1..8451
                     /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"
     promoter        complement(5213..5447)
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     primer_bind     complement(5631..5647)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5632..5648)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(5677..5695)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5716..5732)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5716..5732)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5729..5751)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5740..5756
                     /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(5764..5770,5771..5788,5789..5794))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(5947..5964)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6118..6706)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6198..6217)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6877..7668,7669..7737))
                     /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     7500..7519
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7738..7842)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7879..8382
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     8424..8442
                     /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 gagctctcat ttttgatgaa acagaagctt tttgatattt
     2041 ccatttgaat attttggtat ctgattagtg atgatttcag ctaacatctc tgggaattca
     2101 atactcatgg tcttatccaa aaatgtctgg aagcagtagg taaggagatt ctcaaccacc
     2161 tcatgcatgg agtccagaag ctttgtcagt tggtaaaacc gttgccagtt ctgactggag
     2221 ttcccttccc ttttgacgat ggcttttcct agctctttga tataagtcat tcgaatctca
     2281 tcaaataact cttggctctt cagaccttcc ttaggaactg aggagagaag cagtaaggtt
     2341 ttcatacaga gatactcttc ataggatacc tgcaatcttt gtaattcaga ggagacaaac
     2401 agcatgtgtt tacattggtc atacatgcag ggtagagaca ttctctgctc attaataatc
     2461 agatcaggag caaagcagag caggtttccg cttgattgtc tgtatgatct ccaacccaag
     2521 gcaaatgcca tgagaaacat ccatgagtac tgtagcaggg tcatttggtc atcgaggtgt
     2581 aagtttctca agcctagtat cgcctttgcc catttcactg ctgcaatcac ttgacgccca
     2641 cctaacatgt tgagtgtggt cataattctc catgctgaat ctggaacaga gctatcatat
     2701 cctgcataca acacctcggg ttcaatcacc tccagcagtg acaccaaggt aggggtgagc
     2761 tgtggtaatg ctgcaggaac tattgttttg ttaggatttt ccgaagtgtc ttgtgagact
     2821 cctgcagtgg cttgctgaat ccctttgatt tttttctttg tttttcgagc ttcaaggttc
     2881 attccagcct gaagacattt ccgatagcgg catgctgggc agtttttcct tcgaatttta
     2941 tcaatgatgc aatcgtttct tccagcacaa aggtaattgt gctgtccttc cactgctctt
     3001 ttaaagaata ctttgcagct tccacatgtc agcaccccgt aatgacatcc tgaagcttca
     3061 tcggagcaca ccaggcagag cttgggaggt ggtcccgtgg ctgctgacga gctggatgga
     3121 ggagagctta catctggtct cattccaggg cttgagtacc cattagaaaa cactgaccgg
     3181 cctgggaagt tcagagcccc caaggaagtc aggctgtcct ctccggagcc ttggcaccta
     3241 ttccagtttt cagaaccaac aggaattggt ggaatgacat taaaaacagg cttctgatcc
     3301 tgctgctgag aaagggatgc tgtattcatg tcatagtggt acatctgtcc tccagaggta
     3361 ctcacaccat gaacagaaat ggcagacatt ttattaccaa ttatatttgt cccagaaaag
     3421 cttgcctgac aataaactgg gcccagtttc tcttgcttaa ttaccccggg ggtgcaaagt
     3481 tcaatgaaat catctttttc tgttttcact tggggtagtg ccacactgct gggacttgat
     3541 aagattgtat ctccagtatc cttaatttta ggtttagtgt ccggtaaaat aagaggctta
     3601 caatcctcat tcgtgttccc ttcgagaagg aatggatcat cttctcccgc caaaggagaa
     3661 agcaagtttt catctatcaa cagatctgat ctccagggac tctcgtttgt gtctttactt
     3721 ggggacccag cggaaaactc caaatccttc aagaggtcaa aggtgctttg gtctgtggga
     3781 tacaatttca cactgcctcc gttggtgccg gtctggcttt ttcgattttg ctgttctgaa
     3841 gatgcatccg agtgagtttt gggaaactcc ttctctgtcg gggtagcaca cccagttgca
     3901 gacgttgaac tcttggggtt ctctggaacg ctggtcgacc tattgaggtt tgcaatgctt
     3961 tcttccagaa gccgaaagtc tgtttcccca gaggaaaggc caagttggcc ctgctgtggg
     4021 tagcccaagt cattccccat cacttttgtt tctgtctctc ccatatacag ccccatggac
     4081 agtgaaacgg ctttggataa gcctggctgc tgctgctgct gctgctgctg ctgctgctgc
     4141 tgctgctgct gctgctgctg ctgtcgctgc tgcacattgc ttgtggagcc tttcgagaaa
     4201 tcaaggagaa tcctctgctg cttggaatct gcctgagaag cagcagccac tgagggcgaa
     4261 gatgcagaaa ccttgactgt agctcctccc ctcaggcttt tataaaagtc cattacgctc
     4321 cccctccctt ggccaagcaa actgccaggg acttcgtctc taccaggggg agctaaggat
     4381 tctttggagt ccatccgcgg atcgttaaca acttcgtcat ccacttcttc ttcctgttcc
     4441 ttgatcaagg catcagcact ggtaacattg gcattgttgg tattatgtgt tgattcagag
     4501 attttaaggt cttctacctt tggaagttcc gatttttcct ccgtcttttc ttcagtctgg
     4561 actggtttct cctccttttt aactggttct tcgacctttg ttggctcttt agttggttct
     4621 ttggtttcag cagacttctc ttcctctttc ttatcagatt cggcaggttt tgtgccaacc
     4681 ttcttggtag cattggccaa cttgataccg gaactggaga caagcttcaa agtcttcttt
     4741 ggtttgggag cggcctgctt ttgttgcttt tgaaagtcgt tcaaagacat accttgagac
     4801 tgtggttgga aaccagcttg atatccttgc aaattgttat tgtagttgaa gtttttgtaa
     4861 tttccacggc caccttgtgg attgaattgc tgctgataac cgccttgagg attgtactgt
     4921 tgatagcctc cttgaggatt atactgttgc tggtaaccgg cgtcgggatt gtactgttga
     4981 tagccacctt gttggtaccc agaataacct tggtaatttt ggtagtaccc acctgcaggt
     5041 tgggcttgag cattgtaagc ttgataacct tggtatctgt tgttaccttg ttgttggtta
     5101 ccgttctggc tgtattgctg gtagttttgc tgattgttgc cttggtttga atccgacatg
     5161 gatcctgatt gattgattga ttgtacagtt tgtttttctt aatatctatt tcgatgactt
     5221 ctatatgata ttgcactaac aagaagatat tataatgcaa ttgatacaag acaaggagtt
     5281 atttgcttct cttttatatg attctgacaa tccatattgc gttggtagtc ttttttgctg
     5341 gaacggttca gcggaaaaga cgcatcgctc tttttgcttc tagaagaaat gccagcaaaa
     5401 gaatctcttg acagtgactg acagcaaaaa tgtctttttc taactagtaa caaggctaag
     5461 atatcagcct gaaataaagg gtggtgaagt aataattaaa tcatccgtat aaacctatac
     5521 acatatatga ggaaaaataa tacaaaagtg ttttaaatac agatacatac atgaacatat
     5581 gcacgtatag cgcccaaatg tcggtaatgg gaattcgata tcaagcttat cgataccgtc
     5641 gacctcgagg gggggcccgg tacccagctt ttgttccctt tagtgagggt taattccgag
     5701 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5761 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgaggta
     5821 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5881 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5941 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     6001 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6061 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6121 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6181 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6241 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6301 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6361 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6421 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6481 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6541 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6601 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6661 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6721 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6781 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6841 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6901 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6961 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     7021 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7081 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7141 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7201 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7261 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7321 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7381 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7441 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7501 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7561 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7621 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7681 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7741 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7801 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7861 gccacctggg tccttttcat cacgtgctat aaaaataatt ataatttaaa ttttttaata
     7921 taaatatata aattaaaaat agaaagtaaa aaaagaaatt aaagaaaaaa tagtttttgt
     7981 tttccgaaga tgtaaaagac tctaggggga tcgccaacaa atactacctt ttatcttgct
     8041 cttcctgctc tcaggtatta atgccgaatt gtttcatctt gtctgtgtag aagaccacac
     8101 acgaaaatcc tgtgatttta cattttactt atcgttaatc gaatgtatat ctatttaatc
     8161 tgcttttctt gtctaataaa tatatatgta aagtacgctt tttgttgaaa ttttttaaac
     8221 ctttgtttat ttttttttct tcattccgta actcttctac cttctttatt tactttctaa
     8281 aatccaaata caaaacataa aaataaataa acacagagta aattcccaaa ttattccatc
     8341 attaaaagat acgaggcgcg tgtaagttac aggcaagcga tccgtcctaa gaaaccatta
     8401 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt c
//
