LOCUS       sequence-101201-        3628 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of S. cerevisiae URA3 cloned into pFA6a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3628)
  AUTHORS   Houseley J, Tollervey D
  TITLE     Repeat expansion in the budding yeast ribosomal DNA can occur 
            independently of the canonical homologous recombination machinery.
  JOURNAL   Nucleic Acids Res. 2011 Nov 1;39(20):8778-91. doi: 
            10.1093/nar/gkr589. Epub 2011 Jul 17.
  PUBMED    21768125
REFERENCE   2  (bases 1 to 3628)
  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"; doi: 
            "10.1093/nar/gkr589"; journalName: "Nucleic Acids Res"; date: 
            "2011-11-1- 1"; volume: "39"; issue: "20"; pages: "8778-91"
COMMENT     Sequence Label: pFA6a URA3
FEATURES             Location/Qualifiers
     source          1..3628
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        18..234
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             235..1038
                     /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
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLATGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     primer_bind     complement(1195..1214)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1196..1214)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1301..1318)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1472..2060)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1552..1571)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2231..3022,3023..3091))
                     /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     2854..2873
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3092..3196)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3264..3282
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3320..3342)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3442..3461
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3542..3560
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3542..3559
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gatcctccat aaagcttttc aattcatctt tttttttttt gttctttttt ttgattccgg
       61 tttctttgaa atttttttga ttcggtaatc tccgagcaga aggaagaacg aaggaaggag
      121 cacagactta gattggtata tatacgcata tgtggtgttg aagaaacatg aaattgccca
      181 gtattcttaa cccaactgca cagaacaaaa acctgcagga aacgaagata aatcatgtcg
      241 aaagctacat ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt
      301 aatatcatgc acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag
      361 gaattactgg agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg
      421 gatatcttga ctgatttttc catggagggc acagttaagc cgctaaaggc attatccgcc
      481 aagtacaatt ttttactctt cgaagacaga aaatttgctg acattggtaa tacagtcaaa
      541 ttgcagtact ctgcgggtgt atacagaata gcagaatggg cagacattac gaatgcacac
      601 ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg cggcggaaga agtaacaaag
      661 gaacctagag gccttttgat gttagcagaa ttgtcatgca agggctccct agctactgga
      721 gaatatacta agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt
      781 attgctcaaa gagacatggg tggaagagat gaaggttacg attggttgat tatgacaccc
      841 ggtgtgggtt tagatgacaa gggagacgca ttgggtcaac agtatagaac cgtggatgat
      901 gtggtctcta caggatctga cattattatt gttggaagag gactatttgc aaagggaagg
      961 gatgctaagg tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga
     1021 tgcggccagc aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa
     1081 attagagctt caatttaatt atatcagtta ttacccgggc tgtttaaacg agctcgaatt
     1141 catcgatgat atcagatcca ctagtggcct atgcggccgc ggatctgccg gtctccctat
     1201 agtgagtcgt attaatttcg ataagccagg ttaacctgca ttaatgaatc ggccaacgcg
     1261 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     1321 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     1381 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     1441 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     1501 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     1561 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     1621 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta
     1681 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     1741 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     1801 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     1861 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     1921 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     1981 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2041 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     2101 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     2161 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     2221 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     2281 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     2341 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     2401 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     2461 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     2521 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     2581 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     2641 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     2701 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     2761 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     2821 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     2881 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     2941 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     3001 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     3061 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     3121 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     3181 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     3241 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     3301 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     3361 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     3421 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgg
     3481 acatattgtc gttagaacgc ggctacaatt aatacataac cttatgtatc atacacatac
     3541 gatttaggtg acactataga acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac
     3601 ggatccccgg gttaattaag gcgcgcca
//
