LOCUS       sequence-454889-        3400 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template plasmid for yeast gene knockout, uses auxotrophic marker 
            his5 (Schizosaccharomyces Pombe) (complements S.c. his3).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3400)
  AUTHORS   Feng Y-H, Song J-Z, Zhu J, Xie Z
  TITLE     Template plasmids optimized for deletion of multiple genes in yeast 
            Saccharomyces cerevisiae.
  JOURNAL   Microbiol Spectr. 2024 Oct 10:e0132024. doi: 
            10.1128/spectrum.01320-24.
  PUBMED    39387598
REFERENCE   2  (bases 1 to 3400)
  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: "Microbiol 
            Spectr. 2024 Oct 10:e0132024. doi: 10.1128/spectrum.01320-24."
COMMENT     Sequence Label: SJZ6-HIS5
FEATURES             Location/Qualifiers
     source          1..3400
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..25
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     7..24
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        201..614
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             628..1281
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe his5"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=S. pombe his5
                     /note="yeast auxotrophic marker; corresponds to S. 
                     cerevisiae HIS3"
                     /translation="MGRRAFVERNTNETKISVAIALDKAPLPEESNFIDELITSKHANQ
                     KGEQVIQVDTGIGFLDHMYHALAKHAGWSLRLYSRGDLIIDDHHTAEDTAIALGIAFKQ
                     AMGNFAGVKRFGHAYCPLDEALSRSVVDLSGRPYAVIDLGLKREKVGELSCEMIPHLLY
                     SFSVAAGITLHVTCLYGSNDHHRAESAFKSLAVAMRAATSLTGSSEVPSTKGVL"
     promoter        complement(1650..1668)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(1676..2264)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1756..1775)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2435..3226,3227..3295))
                     /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     3058..3077
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3296..3400)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcattt aggtgacact atagagcggc cgccagctga agcctcgtac gcagctggtc
       61 gacgaatagg cgatggcgct ggtttaatta atcctaggtg aggcgcgcct agggataaca
      121 gggtaataag caacaggcgc gttggacttt taattttcga ggaccgcgaa tccttacatc
      181 acacccaatc ccccacaagt gatcccccac acaccatagc ttcaaaatgt ttctactcct
      241 tttttactct tccagatttt ctcggactcc gcgcatcgcc gtaccacttc aaaacaccca
      301 agcacagcat actaaatttc ccctctttct tcctctaggg tgtcgttaat tacccgtact
      361 aaaggtttgg aaaagaaaaa agagaccgcc tcgtttcttt ttcttcgtcg aaaaaggcaa
      421 taaaaatttt tatcacgttt ctttttcttg aaaatttttt tttttgattt ttttctcttt
      481 cgatgacctc ccattgatat ttaagttaat aaacggtctt caatttctca agtttcagtt
      541 tcatttttct tgttctatta caactttttt tacttcttgc tcattagaaa gaaagcatag
      601 caatctaatc taagttttaa ttacaaaatg ggtaggaggg cttttgtaga aagaaatacg
      661 aacgaaacga aaatcagcgt tgccatcgct ttggacaaag ctcccttacc tgaagagtcg
      721 aattttattg atgaacttat aacttccaag catgcaaacc aaaagggaga acaagtaatc
      781 caagtagaca cgggaattgg attcctggat cacatgtatc atgcactggc taaacatgca
      841 ggctggagct tacgacttta ctcaagaggt gatttaatca tcgatgatca tcacactgca
      901 gaagatactg ctattgcact tggtattgca ttcaagcagg ctatgggtaa ctttgccggc
      961 gttaaaagat ttggacatgc ttattgtcca cttgacgaag ccctttctag aagcgtagtt
     1021 gacttgtcgg gacggcccta tgctgttatc gatttgggat taaagcgtga aaaggttggg
     1081 gaattgtcct gtgaaatgat ccctcactta ctatattcct tttcggtagc agctggaatt
     1141 actttgcatg ttacctgctt atatggtagt aatgaccatc atcgtgctga aagcgctttt
     1201 aaatctctgg ctgttgccat gcgcgcggct actagtctta ctggaagttc tgaagtccca
     1261 agcacgaagg gagtgttgta aatgctatgt aatagacaat aaaaccatgt ttatataaaa
     1321 aaaattcaaa atagaaaacg attctgtaca aggagtattt tttttttgtt ctagtgtgtt
     1381 tatattatcc ttggctaaga ggcactgcgt atacttcaag gtacccctgt gttttgaaaa
     1441 aaaacaacag taaaatagga actccgcgag gttcaggaac ctgaaacaaa atcaataaaa
     1501 acattatatg cgtttcgaac aaaattaaag aaaaagaata aatatagatt aaaaaaaaaa
     1561 agaagaaatt aaaagaattt ctactaaatc ccaattgtta tatatttgtt aaatgccgag
     1621 ctcgatccac tagtggccta tgcccgcggc ctatagtgag tcgtattagt taacttttcc
     1681 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     1741 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     1801 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     1861 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     1921 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     1981 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2041 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2101 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2161 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2221 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     2281 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     2341 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     2401 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     2461 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     2521 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     2581 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     2641 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     2701 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     2761 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     2821 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     2881 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     2941 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     3001 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     3061 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     3121 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     3181 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     3241 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     3301 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     3361 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
//
