LOCUS       sequence-455746-        3738 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template plasmid for yeast gene knockout, confers antibiotic 
            resistance to hygromycin B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3738)
  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 3738)
  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: SJZ5-HPH
FEATURES             Location/Qualifiers
     source          1..3738
                     /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             627..1655
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             627..1655
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(1988..2006)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(2014..2602)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2094..2113)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2773..3564,3565..3633))
                     /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     3396..3415
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3634..3738)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcattt aggtgacact atagagcggc cgccagctga agcttcgtac gccgcaggtc
       61 gacgtatagg agatggtgca ggattaatta accctaggtg aggcgcgcct agggataaca
      121 gggtaataga aaaaaaattt cctggttgaa aatggcggaa aaaaaaaatt cagaaaaaga
      181 aataaagcac gtgtgcgcgg tgtgtggatg atggtttcat cattgtcaac ggcattttcg
      241 ttcttgtgga ttgttgtaaa ctttccagaa cattctagaa agaaagcaca cggaacgttt
      301 agaagctgtc atttgcgttt tttctccaga ttttagttga gaaagtaatt aaattattct
      361 tctttttcca gaacgttcca tcggcggcaa aagggagaga aagaacccaa aaagaagggg
      421 ggccatttag attagctgat cgtttcgagg acttcaaggt tatataaggg gtggattgat
      481 gtatcttcga gaagggattg agttgtagtt tcgtttccca attcttactt aagttgtttt
      541 attttctcta tttgtaagat aagcacatca aaagaaaagt aatcaagtat tacaagaaac
      601 aaaaattcaa gtaaataaca gataatatgg gtaaaaagcc tgaactcacc gcgacgtctg
      661 tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg
      721 gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa
      781 atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatcggccg
      841 cgctcccgat tccggaagtg cttgacattg gggaattcag cgagagcctg acctattgca
      901 tctcccgccg tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg
      961 ttctgcagcc ggtcgcggag gcaatggatg cgatcgctgc ggccgatctt agccagacga
     1021 gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca
     1081 tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca
     1141 gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag
     1201 tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca
     1261 taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca
     1321 acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc
     1381 ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc gtatatgctc cgcattggtc
     1441 ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg
     1501 gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc
     1561 gcagaagcgc ggccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc
     1621 gacgccccag cactcgtccg agggcaaagg aataagttaa ttcaaattaa ttgatatagt
     1681 tttttaatga gtattgaatc tgtttagaaa taatggaata ttatttttat ttatttattt
     1741 atattattgg tcggctcttt tcttctgaag gtcaatgaca aaatgatatg aaggaaataa
     1801 tgatttctaa aattttacaa cgtaagatat ttttacaaaa gcctagctca tcttttgtca
     1861 tgcactattt tactcacgct tgaaattaac ggccagtcca ctgcggagtc atttcaaagt
     1921 catcctaatc gatctatcgt ttttgatagc tcattgagct cgatccacta gtggcctatg
     1981 cccgcggcct atagtgagtc gtattagtta acttttccat aggctccgcc cccctgacga
     2041 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2101 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2161 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2221 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2281 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2341 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2401 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     2461 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2521 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2581 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2641 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     2701 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2761 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2821 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     2881 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     2941 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3001 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     3061 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     3121 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     3181 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     3241 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     3301 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     3361 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     3421 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     3481 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     3541 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     3601 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     3661 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     3721 acaaataggg gttccgcg
//
