LOCUS       sequence-374866-        4758 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gene deletion in S. cerevisiae and S. pombe.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4758)
  AUTHORS   Gadaleta MC, Iwasaki O, Noguchi C, Noma K, Noguchi E
  TITLE     New vectors for epitope tagging and gene disruption in 
            Schizosaccharomyces pombe.
  JOURNAL   Biotechniques. 2013 Nov;55(5):257-63. doi: 10.2144/000114100.
  PUBMED    24215641
REFERENCE   2  (bases 1 to 4758)
  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"; journalName: 
            "Biotechniques"; date: "2013-11"; volume: "55"; issue: "5"; pages: 
            "257-63"
COMMENT     Sequence Label: pFA6a-LEU2MX6
FEATURES             Location/Qualifiers
     source          1..4758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..406
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             407..1501
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(2062..2081)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2063..2081)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2168..2185)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2339..2927)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2419..2438)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3098..3889,3890..3958))
                     /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     3721..3740
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3959..4063)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4131..4149
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4187..4209)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4309..4328
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4409..4427
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4409..4426
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 aactgtggga atactcaggt atcgtaagat gcaagagttc gaatctctta gcaaccatta
       61 tttttttcct caacataacg agaacacaca ggggcgctat cgcacagaat caaattcgat
      121 gactggaaat tttttgttaa tttcagaggt cgcctgacgc atataccttt ttcaactgaa
      181 aaattgggag aaaaaggaaa ggtgagagcg ccggaaccgg cttttcatat agaatagaga
      241 agcgttcatg actaaatgct tgcatcacaa tacttgaagt tgacaatatt atttaaggac
      301 ctattgtttt ttccaatagg tggttagcaa tcgtcttact ttctaacttt tcttaccttt
      361 tacatttcag caatatatat atatatttca aggatatacc attctaatgt ctgcccctaa
      421 gaagatcgtc gttttgccag gtgaccacgt tggtcaagaa atcacagccg aagccattaa
      481 ggttcttaaa gctatttctg atgttcgttc caatgtcaag ttcgatttcg aaaatcattt
      541 aattggtggt gctgctatcg atgctacagg tgttccactt ccagatgagg cgctggaagc
      601 ctccaagaag gctgatgccg ttttgttagg tgctgtgggt ggtcctaaat ggggtaccgg
      661 tagtgttaga cctgaacaag gtttactaaa aatccgtaaa gaacttcaat tgtacgccaa
      721 cttaagacca tgtaactttg catccgactc tcttttagac ttatctccaa tcaagccaca
      781 atttgctaaa ggtactgact tcgttgttgt cagagaatta gtgggaggta tttactttgg
      841 taagagaaag gaagacgatg gtgatggtgt cgcttgggat agtgaacaat acaccgttcc
      901 agaagtgcaa agaatcacaa gaatggccgc tttcatggcc ctacaacatg agccaccatt
      961 gcctatttgg tccttggata aagctaatgt tttggcctct tcaagattat ggagaaaaac
     1021 tgtggaggaa accatcaaga acgaattccc tacattgaag gttcaacatc aattgattga
     1081 ttctgccgcc atgatcctag ttaagaaccc aacccaccta aatggtatta taatcaccag
     1141 caacatgttt ggtgatatca tctccgatga agcctccgtt atcccaggtt ccttgggttt
     1201 gttgccatct gcgtccttgg cctctttgcc agacaagaac accgcatttg gtttgtacga
     1261 accatgccac ggttctgctc cagatttgcc aaagaataag gtcaacccta tcgccactat
     1321 cttgtctgct gcaatgatgt tgaaattgtc attgaacttg cctgaagaag gtaaggccat
     1381 tgaagatgca gttaaaaagg ttttggatgc aggtatcaga actggtgatt taggtggttc
     1441 caacagtacc accgaagtcg gtgatgctgt cgccgaagaa gttaagaaaa tccttgctta
     1501 aaaagattct ctttttttat gatatttgta cataaacttt ataaatgaaa ttcataatag
     1561 aaacgacacg aaattacaaa atggaatatg ttcatagggt agacgaaact atatacgcaa
     1621 tctacataca tttatcaaga aggagaaaaa ggaggatgta aaggaataca ggtaagcaaa
     1681 ttgatactaa tggctcaacg tgataaggaa aaagaattgc actttaacat taatattgac
     1741 aaggaggagg gcaccacaca aaaagttagg tgtaacagaa aatcatgaaa ctatgattcc
     1801 taatttatat attggaggat tttctctaaa aaaaaaaaaa tacaacaaat aaaaaacact
     1861 caatgacctg accatttgat ggagtttaag tcaatacctt cttgaaccat ttcccataat
     1921 ggtgaaagtt ccctcaagaa ttttactctg tcagaaacgg ccttaacgac gtagtcgatc
     1981 gacaagctgt ttaaacgagc tcgaattcat cgatgatatc agatccacta gtggcctatg
     2041 cggccgcgga tctgccggtc tccctatagt gagtcgtatt aatttcgata agccaggtta
     2101 acctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     2161 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2221 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2281 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2341 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2401 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2461 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2521 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2581 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2641 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2701 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2761 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2821 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2881 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2941 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3001 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3061 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3121 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3181 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3241 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3301 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3361 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3421 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3481 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3541 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     3601 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3661 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3721 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3781 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3841 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3901 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3961 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4021 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4081 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     4141 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     4201 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     4261 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     4321 agattgtact gagagtgcac catatggaca tattgtcgtt agaacgcggc tacaattaat
     4381 acataacctt atgtatcata cacatacgat ttaggtgaca ctatagaacg cggccgccag
     4441 ctgaagcttc gtacgctgca ggtcgacgga tccccgggtt aattaaggcg cgccagatct
     4501 tgcaggcatg caagcttgtc gaggagaact tctagtatat ctacatacct aatattattg
     4561 ccttattaaa aatggaatcc caacaattac atcaaaatcc acattctctt caaaatcaat
     4621 tgtcctgtac ttccttgttc atgtgtgttc aaaaacgtta tatttatagg ataattatac
     4681 tctatttctc aacaagtaat tggttgtttg gccgagcggt ctaaggcgcc tgattcaaga
     4741 aatatcttga ccgcagtt
//
