LOCUS       sequence-109932-        4752 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of K. lactis LEU2 cloned into pFA6a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4752)
  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 4752)
  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 LEU2
FEATURES             Location/Qualifiers
     source          1..4752
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2331..2350)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2332..2350)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2437..2454)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2608..3196)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2688..2707)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3367..4158,4159..4227))
                     /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     3990..4009
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4228..4332)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4400..4418
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4456..4478)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4578..4597
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4678..4696
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4678..4695
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 taagcaggct aaccggaacc tgtattattt agtttatgct acgttaaata aagacctttc
       61 gttcacataa ctgaatgtgt aatggccttg agatttcaag cataccaagt tggtggagac
      121 ggggtcgtta caaaagactc tttaagccaa gatttccttg acagccttgg cgatagcatc
      181 gccaacctca gtggtagagt tagaaccacc aaggtcaccg gttctgacac ctgcatccaa
      241 gacatttcta acagcttctt caagagccct accttcttca accaaatcca aggataactt
      301 caacatcata gctgcagata agatggtagc aattgggtta accttgtttg ctggtaaatc
      361 tggggcagaa ccatgacatg gttcgtacaa accgaatgcc ttgttagtgt caggtaggga
      421 agctagagat gcagaaggta ataaacccaa agaacctgga ataacagagg cttcatcgga
      481 gataatatca ccaaacatgt tgttggtaat aacaacaccg tttagcttag ttggtgattt
      541 aaccaaaatc atagcagcag agtcgatcaa ttggtgctga acagttaatt gtgggaactc
      601 agtcttgatg gtttcttcaa cagtctttct ccacaatctg gaagaggcaa gcacgttagc
      661 cttgtcaaga gaccagattg gtaatggtgg gttttgttgc aatgccaaga aagcagccat
      721 tcttgtaatt ctttgaactt caggaacact gtatttctca gagtcccaag caactccgtc
      781 accttcatct tcttttcttt caccaaagta gataccacca accaattctc taacgacgac
      841 gaaatcggta ccctttgcat attcaggctt caaaggagaa agatctagta aagaatcaga
      901 agcaaagtta catggtctca agttggcgta tagacccaat tcctttctga tcttcaatag
      961 accttgttct ggtctaactg cgcccgtacc ccattttgga ccaccaacag cacctagtaa
     1021 gacagcatcg gctttcttag aggcttctag agcttcatct ggtaaaggag tgccagtggc
     1081 atcgatggca gcacccccga tcaagtgatg ttggaaattg aacttaattt ctggacggac
     1141 ttcagcaatg gcattcaaga ccttaatagc ttcgtcagta acttctttac cgacgtgatc
     1201 acccggtagg acaacgatat tcttagacat ggtataatct gtgtagtgtg ggatactttt
     1261 tacttcttca aataggtatc aacaataaaa ataaatcgaa gcaaatgtag gaatgcgtta
     1321 aagcagatgt acttacttag agtacataga tatatttata taattcaata tataaaagta
     1381 tatgaagcat ctgatgttga acctgtcatg actctaaact ggaatgggta gttatgcttc
     1441 tgaaggtttt ccgttagaaa atgagtgcag ggatcaggcc cggaaccggt tttagcctga
     1501 aaggaaaaga aaaaaatgcg gtgattacta gcacgtgact gcgctgaatt ggaatccacc
     1561 acaagtacag cgaatggcaa gagggaaggg gaatatattg tgcctggcgg tagcgtctgc
     1621 tatatcaaat gttaaacata actaaatgca gtagtgaata gagtaatgta tttctacata
     1681 tgtgggtagg gccaatggga gcccgatgtc tattgtacag atattttcct tttatagtta
     1741 atcaattcaa cttggaaatc attgagtcga tatctacttg gataactgga gccgtggcag
     1801 cgtttgaaat ttgacgagtc ttcttgtttt gttcgcgcat tttcagttta ttcaaacggg
     1861 agatggcacc gaatccactg acttctctgt tcaatttgaa tggtaatagg aagaaagtac
     1921 atttcgagaa attttcgccc aacgtcgaaa tctcatggat gatatcctcg atacaaatga
     1981 caccttcatc acctaatttc tcttcaacga tattgttgtc attcaagatt atctcttttg
     2041 gttctttatc ttctggcctt tgccacatta ttctagatct cttttggatt aaagaacgga
     2101 tagatgcaag tgaaggttgt ccgataacaa tatatggtga tacaagtttc aacaatgggt
     2161 acacgtcctt ggtaagtttg atgaaaacac cagtgttcaa ctcaacaagt ctcaaaagag
     2221 ataacacttt gtaagccttt gctgggatct tcacagcgtt taaacgagct cgaattcatc
     2281 gatgatatca gatccactag tggcctatgc ggccgcggat ctgccggtct ccctatagtg
     2341 agtcgtatta atttcgataa gccaggttaa cctgcattaa tgaatcggcc aacgcgcggg
     2401 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2461 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2521 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2581 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2641 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2701 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2761 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     2821 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2881 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2941 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3001 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     3061 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3121 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3181 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3241 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3301 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3361 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3421 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3481 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3541 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3601 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3661 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3721 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3781 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     3841 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     3901 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     3961 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4021 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4081 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4141 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4201 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     4261 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4321 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     4381 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     4441 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     4501 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     4561 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatggacat
     4621 attgtcgtta gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt
     4681 taggtgacac tatagaacgc ggccgccagc tgaagcttcg tacgctgcag gtcgacggat
     4741 ccccgggtta at
//
