LOCUS       sequence-19-expo        7688 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 7688)
  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 7688)
  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-GR
FEATURES             Location/Qualifiers
     source          1..7688
                     /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(4450..4684)
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     primer_bind     complement(4868..4884)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4869..4885)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(4914..4932)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4953..4969)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4953..4969)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4966..4988)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4977..4993
                     /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(5001..5007,5008..5025,5026..5031))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(5184..5201)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5355..5943)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5435..5454)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6114..6905,6906..6974))
                     /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     6737..6756
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6975..7079)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7116..7619
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     7661..7679
                     /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 ccatccggat cctgattgat tgattgattg tacagtttgt ttttcttaat
     4441 atctatttcg atgacttcta tatgatattg cactaacaag aagatattat aatgcaattg
     4501 atacaagaca aggagttatt tgcttctctt ttatatgatt ctgacaatcc atattgcgtt
     4561 ggtagtcttt tttgctggaa cggttcagcg gaaaagacgc atcgctcttt ttgcttctag
     4621 aagaaatgcc agcaaaagaa tctcttgaca gtgactgaca gcaaaaatgt ctttttctaa
     4681 ctagtaacaa ggctaagata tcagcctgaa ataaagggtg gtgaagtaat aattaaatca
     4741 tccgtataaa cctatacaca tatatgagga aaaataatac aaaagtgttt taaatacaga
     4801 tacatacatg aacatatgca cgtatagcgc ccaaatgtcg gtaatgggaa ttcgatatca
     4861 agcttatcga taccgtcgac ctcgaggggg ggcccggtac ccagcttttg ttccctttag
     4921 tgagggttaa ttccgagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     4981 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     5041 gcctaatgag tgaggtaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5101 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5161 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5221 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5281 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5341 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5401 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5461 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5521 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5581 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5641 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5701 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5761 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     5821 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5881 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5941 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6001 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6061 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6121 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6181 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6241 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6301 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6361 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6421 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6481 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6541 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6601 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6661 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6721 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6781 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6841 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6901 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6961 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7021 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7081 acatttcccc gaaaagtgcc acctgggtcc ttttcatcac gtgctataaa aataattata
     7141 atttaaattt tttaatataa atatataaat taaaaataga aagtaaaaaa agaaattaaa
     7201 gaaaaaatag tttttgtttt ccgaagatgt aaaagactct agggggatcg ccaacaaata
     7261 ctacctttta tcttgctctt cctgctctca ggtattaatg ccgaattgtt tcatcttgtc
     7321 tgtgtagaag accacacacg aaaatcctgt gattttacat tttacttatc gttaatcgaa
     7381 tgtatatcta tttaatctgc ttttcttgtc taataaatat atatgtaaag tacgcttttt
     7441 gttgaaattt tttaaacctt tgtttatttt tttttcttca ttccgtaact cttctacctt
     7501 ctttatttac tttctaaaat ccaaatacaa aacataaaaa taaataaaca cagagtaaat
     7561 tcccaaatta ttccatcatt aaaagatacg aggcgcgtgt aagttacagg caagcgatcc
     7621 gtcctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     7681 ctttcgtc
//
