LOCUS       sequence-454888-        3536 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template plasmid for yeast gene knockout, confers antibiotic 
            resistance to G418.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3536)
  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 3536)
  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: SJZ7-KAN
FEATURES             Location/Qualifiers
     source          1..3536
                     /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"
     CDS             644..1453
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(711..730)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1786..1804)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(1812..2400)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1892..1911)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2571..3362,3363..3431))
                     /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     3194..3213
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3432..3536)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcattt aggtgacact atagagcggc cgccagctga agcatcgtac gccgctggtc
       61 gacggatcgg tgacggtgcc ggcttaatta accctaggtg aggcgcgcct agggataaca
      121 gggtaatgca cgcgcgatgg tgctaagtta catatatata tatatatata tatatatata
      181 tatagccata gtgatgtcta agtaaccttt atggtatatt tcttaatgtg gaaagatact
      241 agcgcgcgca cccacacaca agcttcgtct tttcttgaag aaaagaggaa gctcgctaaa
      301 tgggattcca ctttccgttc cctgccagct gatggaaaaa ggttagtgga acgatgaaga
      361 ataaaaagag agatccactg aggtgaaatt tcagctgaca gcgagtttca tgatcgtgat
      421 gaacaatggt aacgagttgt ggctgttgcc agggagggtg gttctcaact tttaatgtat
      481 ggccaaatcg ctacttgggt ttgttatata acaaagaaga aataatgaac tgattctctt
      541 cctccttctt gtcctttctt aattctgttg taattacctt cctttgtaat tttttttgta
      601 attattcttc ttaataatcc aaacaaacac acatattaca ataatgggta aggaaaagac
      661 tcacgtttcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
      721 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgattgtatg ggaagcccga
      781 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
      841 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
      901 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggcaaaa cagcattcca
      961 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     1021 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     1081 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     1141 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataagc ttttgccatt
     1201 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     1261 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     1321 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     1381 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     1441 tgagtttttc taagcgattt aatctctaat tattagttaa agttttataa gcatttttat
     1501 gtaacgaaaa ataaattggt tcatattatt actgcactgt cacttaccat ggaaagacca
     1561 gacaagaagt tgccgacagt ctgttgaatt ggcctggtta ggcttaagtc tgggtccgct
     1621 tctttacaaa tttggagaat ttctcttaaa cgatatgtat attcttttcg ttggaaaaga
     1681 tgtcttccaa aaaaaaaacc gatgaattag tggaaccaag gaaaaaaaaa gaggtatcct
     1741 tgattaagga acagagctcg atccactagt ggcctatgcc cgcggcctat agtgagtcgt
     1801 attagttaac ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     1861 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     1921 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     1981 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     2041 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2101 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     2161 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     2221 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     2281 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     2341 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     2401 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     2461 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     2521 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     2581 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     2641 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     2701 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     2761 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     2821 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     2881 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     2941 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     3001 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     3061 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     3121 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     3181 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     3241 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     3301 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     3361 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     3421 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     3481 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcg
//
