LOCUS       sequence-453990-        3890 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template plasmid for yeast gene knockout, uses auxotrophic marker 
            LEU2 (Kluyveromyces lactis).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3890)
  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 3890)
  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: SJZ10-LEU2
FEATURES             Location/Qualifiers
     source          1..3890
                     /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        complement(2140..2158)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(2166..2754)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2246..2265)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2925..3716,3717..3785))
                     /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     3548..3567
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3786..3890)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcattt aggtgacact atagagcggc cgccagctga agcatcgtac gccgcaggtc
       61 gacgaattgg agacggtgct ggattaatta atcctaggtg aggcgcgcct agggataaca
      121 gggtaatctc gtcaaatttc aaacgctgcc acggctccag ttatccaagt agatatcgac
      181 tcaatgattt ccaagttgaa ttgattaact ataaaaggaa aatatctgta caatagacat
      241 cgggctccca ttggccctac ccacatatgt agaaatacat tactctattc actactgcat
      301 ttagttatgt ttaacatttg atatagcaga cgctaccgcc aggcacaata tattcccctt
      361 ccctcttgcc attcgctgta cttgtggtag attccaattc agcgcagtca cgtgctagta
      421 atcaccgcat ttttttcttt tcctttcagg ctaaaaccgg ttccgggcct gatccctgca
      481 ctcattttct aacggaaaac cttcagaagc ataactaccc attccagttt agagtcatga
      541 caggttcaac atcagatgct tcatatactt ttatatattg aattatataa atatatctat
      601 gtactctaag taagtacatc tgctttaacg cattcctaca tttgcttcga tttattttta
      661 ttgttgatac ctatttgaag aagtaaaaag tatcccacac tacacagatt ataccatgtc
      721 taagaatatc gttgtcctac cgggtgatca cgtcggtaaa gaagttactg acgaagctat
      781 taaggtcttg aatgccattg ctgaagtccg tccagaaatt aagttcaatt tccaacatca
      841 cttgatcggg ggtgctgcca tcgatgccac tggcactcct ttaccagatg aagctctaga
      901 agcctctaag aaagccgatg ctgtcttact aggtgctgtt ggtggtccaa aatggggtac
      961 gggcgcagtt agaccagaac aaggtctatt gaagatcaga aaggaattgg gtctatacgc
     1021 caacttgaga ccatgtaact ttgcttctga ttctttacta gatctttctc ctttgaagcc
     1081 tgaatatgca aagggtaccg atttcgtcgt cgttagagaa ttggttggtg gtatctactt
     1141 tggtgaaaga aaagaagatg aaggtgacgg agttgcttgg gactctgaga aatacagtgt
     1201 tcctgaagtt caaagaatta caagaatggc tgctttcttg gcattgcaac aaaacccacc
     1261 attaccaatc tggtctcttg acaaggctaa cgtgcttgcc tcttccagat tgtggagaaa
     1321 gactgttgaa gaaaccatca agactgagtt cccacaatta actgttcagc accaattgat
     1381 cgactctgct gctatgattt tggttaaatc accaactaag ctaaacggtg ttgttattac
     1441 caacaacatg tttggtgata ttatctccga tgaagcctct gttattccag gttctttggg
     1501 tttattacct tctgcatctc tagcttccct acctgacact aacaaggcat tcggtttgta
     1561 cgaaccatgt catggttctg ccccagattt accagcaaac aaggttaacc caattgctac
     1621 catcttatct gcagctatga tgttgaagtt atccttggat ttggttgaag aaggtagggc
     1681 tcttgaagaa gctgttagaa atgtcttgga tgcaggtgtc agaaccggtg accttggtgg
     1741 ttctaactct accactgagg ttggcgatgc tatcgccaag gctgtcaagg aaatcttggc
     1801 ttaataagag taataattat tgcttccata taatattttt atatacctct tatttttatg
     1861 tattagttaa ttaagtattt ttatctatct gcttatcatt ttcttttcat ataggggggg
     1921 ttggtgtttt cttgcccatc agattgatgt cctccaactc ggcactattt tacaaagggt
     1981 ttttttgtaa gagaaggaga agacagatac taaaccatac gttactcgaa acaaaaaaaa
     2041 aaaaaatgga aaaagctgct atcaacaaaa gacggcctca tcaaacctaa agaaaccatg
     2101 tcagcgtgag ctcgatccac tagtggccta tgcccgcggc ctatagtgag tcgtattagt
     2161 taacttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2221 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2281 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2341 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2401 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2461 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2521 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2581 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2641 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2701 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2761 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     2821 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2881 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     2941 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3001 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3061 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3121 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3181 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3241 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3301 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3361 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3421 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3481 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3541 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     3601 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3661 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     3721 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     3781 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     3841 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
//
